On Methods Of Analysis And Sampling And Preparation Of Control Samples

Original Language Title: o metodách zkoušení a způsobu odběru a přípravy kontrolních vzorků

Read the untranslated law here: https://portal.gov.cz/app/zakony/download?idBiblio=57811&nr=211~2F2004~20Sb.&ft=txt

211/2004 Coll.
DECREE


Dated 15 April 2004

On methods of analysis and sampling and preparation of control samples

Change: 611/2004 Coll.

Change: 238/2005 Coll.

Change: 459/2005 Coll.

The Ministry of Agriculture stipulates pursuant to § 18 point. n) of the Act no. 110/1997
Coll., on foodstuffs and tobacco products and amending and supplementing
some related laws, as amended by Act no. 119/2000 Coll., Act No.
. 306/2000 Coll., Act no. 146/2002 Coll. and Act no. 274/2003 Coll., (hereinafter
"Act").

§ 1
General


(1) This decree ^ 1) transposes the relevant European Community
^ 1) and provides testing and sampling and preparation
control samples (hereinafter "Samples") for the purpose of determining the quality and || | food safety and quality of tobacco products within
state supervision, with the exception of sampling for microbiological testing.

(2) When testing, sampling and sample preparation of food or tobacco products
may be used other scientifically valid methods (such as methods
International Organization for Standardization, or Codex Alimentarius)
provided, however, that their use is not an obstacle to the free movement of goods
. In the event that there are discrepancies in the results of testing are considered to be determinant
test results obtained using the methods outlined in
hereto.

(3) For food and tobacco products, which do not by this Decree
defined test methods or not use specified in this Decree
sampling and preparation of the samples, shall be governed by
paragraph 2, first sentence similarly .

§ 2
Basic concepts


For the purposes of this Decree

A) part of the batch ^ 2) - defined part of a lot in order to apply certain methods
taking the sample, with each part of the batch is
physically separate and separately identifiable, unless stated otherwise
, | ||
B) controlled dose - defined quantity of food or tobacco
product, which as a whole is presented for inspection; It may be a
lot or a part of a lot,

C) of the sampled unit - one of the basic units each of which is composed
basic file or quantity of food or tobacco product
forming a coherent unit and taken suddenly from one place
so that formed part of subsample ; a basic set is considered
file all considered sampled units

D) sample - one or more of the sampled units taken from
basic set and intended to enable them to obtain information about the basic
file

E) sub-sample - the quantity of material taken from a single place in the lot or sublot
,

F) the aggregate sample - sample composed of all the incremental samples

G) reduced sample - the aggregate sample or a representative portion
prepared from the bulk sample separation; reduced sample
prepared only in case where the aggregate sample is too large,

H) laboratory sample - sample intended for laboratory tests

I) duplikátním sample - one of the two or more samples obtained at
same time using the same sampling procedure and sample division,

J) dividing the sample - the technique to take one or more of the subsamples
sample nekusové food or tobacco product by means such as cutting
lattice divider, mechanical separation or separation

K) sub-sample - sample taken from the population;
nekusové when ordering food or tobacco product subsamples are prepared
dividing samples

L) sample preparation - summary of the operations with the foodstuff or tobacco
product, such as size reduction, blending, division,
necessary to convert the aggregate sample to the laboratory sample

M) sampling (sampling) - The process of sampling,

N) the sampling rate - the number of units subscribed sampled,

O) random sampling - sampling the same level
probability that a random sample of the batch checked will be selected,

P) targeted sampling - sampling for a specific purpose
unequal level of confidence that any sample will be selected,

Q) sampling of nekusové food or tobacco product - taking
sample nekusové food or tobacco products submitted in

Doses, which can not immediately distinguish the sampled unit,

R) multistage sampling - sampling in which the sample
form by degrees so that the unit in each stage is removed from the larger
units selected in the previous step,

S) laminated sampling - sampling executed from the basic
file which can be divided into mutually exclusive subsets and
covering is such that the specified portion of the sample is removed from
different layers from each layer is removed at least one unit,

T) sampling plan - a plan setting out the scope of sampling

U) test report - a document setting forth the results of laboratory tests set
requirements

In) accuracy - closeness of agreement between a test result and the accepted
benchmark

W) accuracy - closeness of agreement between the average value obtained from a large
series of test results and an accepted reference value

X) commonality - the closeness of agreement between independent test results obtained under
prespecified conditions

Y) repeatability conditions - conditions where independent test results are obtained
same test method on identical test
units in the same laboratory by the same operator, using the same equipment
during the short time span ,

A) repeatability - precision under repeatability conditions,

Aa) reproducibility - precision under reproducibility conditions,

Bb) reproducibility conditions - the conditions under which the test results
receive the same test method on identical test
units in different laboratories, different operators that use
variety of amenities

Cc) test result - the character value obtained by performing the method
testing

Dd) errors in the results - test result minus the accepted reference value
character

Ee) specificity - characterization testing methods stipulating for
which features the method is specific,

Ff) the detection limit - the lowest value that can be determined by the
testing

Gg) fitness for use - characteristics which define methods for testing purpose
for which the test method specified,

Hh) usability - characterization test methods specifying
for what purposes the method is applicable testing,

Ii) the accuracy of the experiment - an interlaboratory test suitability
appropriate test methods.
Sampling


§ 3

(1) Sampling by the person authorized for this activity and
properly trained. ^ 3)

(2) The sample is first detected

A) batch designation as indicated on the packaging of food intended for
consumer or the transport packaging and documents relating to the
food,

B) the weight or volume of the lot or the number of units in the lot or
controlled entity

C) the type and size of packaging and labeling

D) the possible presence of parts of the lot

First corrupt, damaged, or otherwise objectionable; These are part of the batch
off a sample for testing from them does not remove, if not the
suspected food when the procedure under paragraph 12

Second safe; of each homogeneous part of the batch to be sampled separately
.

(3), before the start of sampling down

A) sampling procedure or if it is already set, the sample collection process
specify

B) the required types of tests

C) sampling plan,

D) the extent of sampling for each test by sampling plan,

E) total mass or volume of sample required to perform all
required inspections and laboratory tests, the combination of the range of sampling and sample
kinds of tests

F) the total required number of packages or partial samples that are collected
.

(4) The acceptance plan shall be determined in accordance with the procedures outlined in the Czech
technical standards governing the acceptance procedures, statistical
acceptance and more details ^ 4) or in accordance with procedures processed
supervisory authority and disclosed website
supervisory authority.

(5) The sample is taken from each batch separately controlled so that
always represent the whole lot, unless stated otherwise.

(6) The sample consists of one or more sub-samples.
Sub-sample is taken from a randomly chosen point in the lot; if this is not possible,

Collected from randomly selected sites in accessible parts of the lot.
Subsample size is calculated as the proportion between the desired size
general laboratory sample or a set number of units subscribed
.

(7) fractional sampling is performed

A) by random sampling, in which all the sampled units have the same probability
removal,

B) sampling the in which each sampled unit
taken at predetermined distances or intervals
randomly chosen from the beginning, or

C) the manner in which each sampled unit
taken from different locations.

(8) Partial samples taken from the packaged food without damaging the packaging
intended for consumers, a non-prepacked food shall be taken
partial samples in violation of the outer packaging.

(9) liquid or semi-liquid food in storage containers
sub-sample taken after mixing the contents. If you can not mix the contents,
removes a sub-sample of the individual layers of stratified sampling.
Similarly for dry foodstuffs or unpackaged food, where
sub-sample taken from different layers (strata) or from various locations
appropriate sampling equipment.

(10) In addition, partial sample remove

A) of unpackaged food sampling device or odkrojením,

B) of unpackaged foods consisting of solid and liquid substances independently of
solid and liquid components, or

C) fresh fruit, fresh vegetables, potatoes and fresh mushrooms
.

(11) In the case that the food is simultaneously in multiple containers
particular in any outer packaging used multistage sampling
sample so that it is selected

A) in the first stage of primary sample, which is transport packaging

B) in the second stage of secondary sample, which is packing
removed from the shipping container,

C) sample in subsequent stages similarly under point b), so that in
last step was taken subsample of the package intended for consumers
.

(12) U suspect foods ^ 2) is carried out targeted sampling used for
laboratory testing to detect faults in suspect foods. Part of the batch in question
food must be specified prior to sampling.
This sample is not representative of the whole lot and have to be labeled
.

(13) The sample must be taken such precautionary measures
to prevent its depreciation and any changes that would affect the
testing methods.

(14) Frozen food must not thaw during sampling.

(15) The sampling referred to in paragraphs 1 to 13 also apply to
sample evaluation on the spot.

§ 4

(1) When sampling of tobacco products is progressing in accordance with the procedures set forth in
Czech technical standards governing
taking samples of tobacco products. ^ 5)

(2) If you can not at a sampling of tobacco products to follow
paragraph 1 shall be used for the sampling of tobacco provisions of §
3 appropriately.

(3) When sampling for the determination of aflatoxins in foods
samples for determination of pesticides in food and their
surface, samples of tea, samples of cereals, legumes and grain mill products, animal samples
and vegetable fats and oils and control samples
temperature of frozen food the procedure set out in the Czech
technical standards for the collection of samples and testing methods
certain foods. ^ 6)

(4) When sampling of dried milk products, dairy products
thickened, casein and caseinates the procedure set out in
Czech technical standards governing the testing methods
dairy products, casein and caseinates. ^ 7 )

(5) When sampling food for inspection ochratoxin A, samples for
control of dioxins and the determination of dioxin-like polychlorinated biphenyls
PCBs in certain foodstuffs and samples for compliance levels of lead,
cadmium, mercury and 3 -chlorpropan-1,2-diol as described in Annex no. 1 to 3.


(6) When sampling of raw and heat-treated milk
proceed in accordance with the procedures laid down in European Community regulations. ^ 8)

(7) When sampling fresh fruit and vegetables
proceed in accordance with the procedures laid down in European Community regulations. ^ 9)

(8) In Sampling of olive oil should be followed in

Accordance with the procedures laid down in European Community regulations. ^ 10)

(9) When sampling for control of tin content in foods packaged in cans
as described in Annex no. 40.

(10) When sampling for control of the levels of patulin in foodstuffs is
described in Annex no. 41.

(11) Sampling of benzo [a] pyrene in foodstuffs
as described in Annex No. 44.

(12) When sampling for the detection of genetically modified organisms and
material produced from genetically modified organisms or products containing them
into account the recommendations set out in the Regulation of the European Communities ^
10a).

(13) When sampling to check the content of Fusarium toxins in food
as described in Annex no. 46.

§ 5

Sampling protocol

(1) of sampled Each sample must be drawn up a protocol for sampling
sample, which allows unique identification of controlled
food or tobacco product, the lot or sublot.

(2) the sampling protocol must contain

A) protocol number,

B) the data referred to in § 6 para. 1 point. a) of the Act or identifier
food business operator

C) the name of the food or tobacco product under which it is put into circulation
,

D) an indication of the amount of food or tobacco product in the package (volume
weight or number of pieces)

E) indication of the lot:

First batch designation under § 3 para. 2 point. a)

Second the extent or size of the sampled lot; for non-prepacked food
number of packing units or their weight in food nekusové
total mass or volume,

Third production date, if indicated

Fourth expiration date or date of minimum durability;

F) information on sampling:

First a reference to the Czech technical standard, or deviations from the used
Czech technical standards, or a reference to a decree

Second details of any environmental conditions during sampling that may affect
test results

Third instead of sampling or graphs, drawings or photographs,

Fourth date and time of sampling,

Fifth the purpose of sampling,

6th the amount of sample for laboratory testing; the number and amount of
package for non-prepacked food, weight or volume at nekusové foods

7th Name and signature of the person who performed the sampling, and signature
controlled person;

G) information for the laboratory, which may affect the quality and safety of health
, especially about the time of shipment of the sample, the conditions under which it was
sampling and any suspected violations
quality or health,

H) other data, including in particular the type of packaging sample
way to ensure the integrity of the sample used sampling devices or other circumstances
during sampling, which could affect the assessment
sample, the state-controlled food or tobacco product,
possible presence spoiled, contaminated or otherwise defective parts
batch and sampling of these parts of the batch

I) information on whether the duplicate sample was taken.

§ 6

Packaging, labeling and transport of sample

(1) Each sample is placed in a clean and inert container which protects
sample from contamination and damage during transportation. At the same time
implement the necessary measures to avoid any change in composition of the sample,
that might occur during transport.

(2) The sample packing containers are used compliant
special legal regulation, 11) which do not affect the results of laboratory tests
.

(3) The sample must be delivered to the laboratory as soon as possible. When transporting
not cause deterioration. A sample of frozen food must
remain permanently frozen sample and perishable food
permanently chilled or frozen.

(4) The sample shall be marked, closed and sealed so as to avoid confusion
sample and to open the package without damaging the packaging or seals.

(5) The sample is labeled with relevant information about

A) the name of the product

B) batch under § 3 para. 2 point. a)

C) the sampling protocol,

D) other facts about the method of sampling, if
could affect the test results.

(6) If you can not sample mark pursuant to paragraph 5, a sample can indicate
just an indication pursuant to paragraph 5. C).

(7) The sample must be delivered and handed over to the lab immediately after his

Collection. If there is a sample during transport under official control, the person
that performed the sampling, ensure that there
during transport to damage the sample.

(8) In the event that you can not sample immediately after it is removed
sent to the laboratory, the person who performed the sampling, such
measures to ensure that the sample taken prior to dispatch stored under conditions || | where there is no visible damage to the sample and confusion. This does not apply to samples
perishable foodstuffs or frozen specimens
food.

§ 7
Sample preparation


(1) When preparing the sample used

A) homogenizing, such as mixing, stirring, and the reduction of grain, crushing, grinding
,

B) division, for example, reducing the sample to a divider, cutting, slicing or
quartering; while quartering means the elimination of two opposite quarters, mixing and re
quartering balance until it reaches the desired size
or

C) the combination of homogenization and division.

(2) During the preparation of laboratory samples, such precautionary measures
to avoid any change that would affect the outcome
test.

(3) merging and mixing all subsamples are prepared
aggregate sample, unless otherwise specified. If necessary the aggregate sample
modify the manner specified in paragraph 1, or from the prepared
reduced sample.

(4) The aggregate sample is prepared laboratory sample and a duplicate sample
. Laboratory sample was labeled in a manner allowing its
unambiguous identification. In order to quantity the analyte
calculated to include parts which are not analyzed, the mass separated
portion is recorded.

(5) Laboratory sample as required pulverized and mixed well to
order to collect a representative test portions. Size of testing
shares is determined by testing and mixing efficiency.
Methods of grinding and mixing must not affect the composition of the test portion.
Test portion is treated as necessary under specific conditions in order to minimize the adverse effects
.

(6) Replicate sample shall bear a manner allowing its unambiguous identification
and stored for retesting or additional tests.
Method and duration of storage of a duplicate sample shall not affect its
composition.

(7) Using a sample for laboratory analyzes expires
date of application or date of minimum durability, which would occur at a time after his
subscribe to the commencement of laboratory tests.

(8) when preparing samples for testing animal and vegetable fats and oils
proceed according to Czech technical standards governing the procedure for the preparation of samples for
certain fats and oils. ^ 12)

(9) In the preparation of samples for determining the levels of ochratoxin A,
samples for determination of lead, cadmium, mercury and 3-chloro-propane-1,2-diol in certain
foods and samples for determination of dioxins and polychlorinated biphenyls
dioxin-like PCBs in certain foodstuffs as described in Annex
no. 5 to 7

(10) When preparing samples for olive oil
proceed in accordance with the Regulation of the European Communities. ^ 10)

(11) Laboratory samples can be prepared on-site sampling,
only if it is not influenced by the composition of the sample, and it will not be
devaluation. When preparing the sample shall be governed by paragraphs 1 to 10

(12) When preparing samples for tin content in foods packaged in cans
as described in Annex no. 42.

(13) when preparing samples for checking compliance with maximum levels of patulin in foodstuffs
as described in Annex no. 43.

(14) When preparing samples for levels of benzo [a] pyrene in
foods as described in Annex no. 45.

(15) When preparing samples for the detection of genetically modified organisms
and material produced from genetically modified organisms or products
their contents into account the recommendations set out in the European Community regulations
^ 10a).

(16) When preparing samples for the content of Fusarium toxins in food
as described in Annex no. 47.
Testing methods


§ 8

(1) Sensory evaluation performed by a person authorized for this activity and properly trained
^ 3) in accordance with the requirements of Czech technical standards
governing procedure and training of assessors. ^ 13)


(2) When the sensory evaluation progresses person referred to in paragraph 1
according to Czech technical standards governing sensory analysis. ^ 14)

(3) Inspection of the sensory properties of olive oil is
procedure set down in European Community. ^ 10)

§ 9

(1) Quality control and food safety and quality
tobacco products is carried out on samples taken in accordance with § 3 and 4
appropriate testing methods.

(2) preferred to use testing methods that are applicable
same way for different groups of food or tobacco products before
testing methods that are applicable only to some
food or tobacco product.

(3) For each test methods to be used for official control and
that's its purpose and nature does not rule must be set at least
following characteristics:

A) specificity,

B) precision

C) consistency, repeatability and reproducibility,

D) limit of detection,

E) usability and usability.

(4) Methods of testing must be arranged in the form of recommendations for testing methods
International Organization for Standardization. ^ 15)

(5) The exact values ​​are obtained by evaluating the conformity experiments
accuracy, which took place in accordance with the procedures outlined in the Czech
technical standards governing the accuracy of measurement methods and results. ^ 16)

(6) repeatability and reproducibility values ​​shall be expressed in the form
referred to in technical standards ^ 16) while the normal value
probability level is 95%.

(7) Results of experiments, the accuracy of the supervisory authority shall publish on its website
.

(8) Methods of testing laboratories perform supervisory authorities. ^ 3)
The implementation methods of analysis for the official control may also be charged
other laboratories that meet the requirements of special legal regulation
^ 17) and Czech technical standards governing requirements
lab. ^ 17)

§ 10

(1) For cigarettes inspection of tar, nicotine and carbon monoxide
evaluate the accuracy of the tar and nicotine numbers on the packages
intended for consumers carried out by the Czech technical standards governing
cigarettes. ^ 18 )

(2) Checking the physical and chemical characteristics of olive oils and their composition is done
methods specified in regulations of the European
communities. ^ 10)

(3) Inspection of the physical and chemical properties of the casein and caseinates are
carried out by the Czech technical standards governing the testing methods
casein and caseinates. ^ 19)

(4) Control of dried milk products and dairy thickened
is done according to Czech technical standards governing the testing methods
dairy products. ^ 20)

(5) When checking the temperature of frozen foods is governed by Czech
technical standards governing the testing methods frozen products. ^ 21)

(6) Checking the physical and chemical quality characteristics for spirits is done
methods set out in European Community regulations. ^ 22)

(7) In determining the value of refractive solids
proceed in accordance with the procedure laid down in European Community regulations. ^ 23)

(8) In determining the content of starch and its fission products including
glucose determination of starches or dextrins or other modified starches
proceed in accordance with the methods outlined in
European Community regulations. ^ 24)

(9) When checking the contents of ochratoxin A as described in Annex no. 5

(10) When checking the levels of lead, cadmium, mercury and
3-chloro-propane-1,2-diol in certain foods is governed by
Annex no. 6.

(11) When checking the levels of dioxins and polychlorinated biphenyls
determination of dioxin-like PCBs in certain foodstuffs as described in Annex
no. 7th

(12) When checking the level of erucic acid in fats and oils and
foods made from them are described in Annex No. 11.

(13) When checking the physical and chemical characteristics of certain sugars
proceed in accordance with Annexes Nos. 12-22 and annex no. 39.

(14) When checking the freezing point, phosphatase and peroxidase activity in
raw and heat-treated milk is progressing in accordance with the methods
testing specified in the Regulation of the European Communities. ^ 8)

(15) while checking the purity of additives in food proceed
according to Annex no. 23 to 38.


(16) When checking the water content of solids-non-fat butter
proceed in accordance with the testing methods set out in the regulations of the European
communities. ^ 25)

(17) The quality inspection of fresh fruit and vegetables
proceed in accordance with the testing methods set out in the regulations of the European
communities. ^ 8)

(18) When checking the physical, chemical and sensory characteristics of wine
governed by special legislation. ^ 26)

(19) When checking samples of tin content in foods packaged in cans
as described in Annex no. 42.

(20) when checking compliance with maximum levels of patulin in foodstuffs
as described in Annex no. 43.

(21) When checking the levels of benzo [a] pyrene in foodstuffs proceed
according to Annex no. 45.

(22) Upon detection of genetically modified organisms and material
produced from genetically modified organisms or their products
contents into account the recommendations set out in the regulations of the European Communities ^
10a).

(23) When checking the content of Fusarium toxins in food
proceed in accordance with Annex no. 47.

§ 11
Expressing results


(1) Results of laboratory tests on the quality and health
audited sample shall be included in the test report, which must
contain the information necessary for the representation of test results and information
required testing method used.

(2) The test report must include at least the following information:

A) protocol number,

B) the name and address of the laboratory and the place where the tests were performed, if
these data are different from the address of the laboratory

C) identification of the test report; each page of the test report
must be identifiable as part of the test report and be
apparent end of a test,

D) the business name or name, manufacturer, importer, seller
or packager, and its registered office, if a legal person, and
permanent residence or place of business, if a natural person ,

E) the method used for testing

F) clear identification of test performed,

G) the date of receipt of the sample, if it is important for the use of force and
results, and the date of testing,

H) a reference to the sampling plan and sampling procedure according to § 3 and 4 or
link protocol for sampling pursuant to § 5

I) the results of the test and measurement unit by
Czech technical standards ^ 27)

J) the name, title and signature of the person receiving the test report,

A) a statement that the test result applies only to the test
samples,

L) the number of pages and the total number of pages in the printed output protocol
a test

M) a statement that the test report may not be reproduced
laboratory in which the test was performed, published other than whole,

N) deviations, additions or exclusions relating to methods of testing and
information on specific test conditions, such conditions
environment

A) a statement of compliance or non-compliance with the requirements of the testing methods, if
is necessary

P) a statement of the estimation of measurement uncertainty, if necessary;
information on the measurement uncertainty is always required if the uncertainty affect the value
compliance with the relevant test methods

Q) any expert opinions and expressions used in the implementation
tests in accordance with the Czech technical standard governing the requirements
testing and calibration laboratories, ^ 28)

R) any additional information that may be required for methods
testing.

(3) The test result is the average of the results of at least two parallel
determination, unless otherwise specified. Part of the results of the test must be always
error result.

(4) Upon learning that the test result exceeds the limit
identified substances shall immediately be repeated examination to confirm
previously obtained result, and using scientifically proven methods
in accordance with § 1. 2 or 3.

(5) Test results are shown with or without correction for recovery.
If it is mentioned correction for recovery shall indicate in the report
test recoveries. Yield value is the ratio of the detected
amount of substance in the sample and the actual, known or
added amount of the substance in the sample and is expressed in percentage.

§ 12
Repealing provisions


The Decree no. 339/2001 Coll., On methods of analysis and

Sampling and preparation of the samples in order to measure the quality and
health food or raw materials for their production and
quality tobacco products.

§ 13
Efficiency


This Decree shall take effect on the date of the accession
Czech Republic to the European Union.
Minister
:

Ing. Palas vr


Příl.1
Sampling procedure for determining the levels of ochratoxin A in
certain foodstuffs and raw materials

First

Purpose and scope

Samples intended for official control of the levels of ochratoxin A in foods must be taken
methods described below. Thus obtained aggregate
samples are considered representative of the lots.
Compliance with maximum limits laid down in the European Community regulations no. 466/2001
will be determined based on the amount determined in the laboratory samples
.

Second


Definition
Lot: an identifiable quantity of food delivered at one time
which holds an official determination
uniform characteristics such as origin, variety, type of packing, packer, consignor or markings
.

Part of the batch: a certain part of a large lot allocated to it was
sampled. Each sublot must be physically separate and identifiable
.

Incremental sample: a quantity of material taken from a single place in the lot or sublot
.

A comprehensive summary of all the incremental samples taken from the lot sample:
or part of the batch.

Third
General

3.1
Staff

Sampling must be performed by an authorized officer (§ 3 para. 1 of this Decree
).

3.2 Material to be

Each lot which is to be examined must be sampled separately.
Large lots under this Annex is broken into sections to be sampled separately
.

3.3 Precautions

Of sampling and preparation of the samples must be taken
precautions to avoid any changes, which would affect
ochratoxin A content, adversely affect the analytical determination or
aggregate samples unrepresentative.
Sub-samples 3.4


Incremental samples shall, if possible, from various places throughout the lot or sublot
. Deviations from this procedure must be recorded in the log.

3.5 Preparation of aggregate sample

The aggregate sample is made up by uniting the incremental samples.

3.6 Samples for re-examination

Samples for repeated tests to confirm, defense
commercial dispute or referee purposes shall be taken from the laboratory sample
if it does not conflict with Member States' rules on sampling
samples.

3.7 Packaging and transmission of laboratory samples

Each sample is placed in a clean, inert container which
provides adequate protection from contamination and damage during transportation
. They must be taken all the necessary measures to prevent
change in composition of the sample, which might arise during transportation or storage.

3.8 Sealing and labeling of samples

Each sample taken for official controls sealed at the place
sampling and identified according to § 6 of this Decree. For each sample must be exposed
protocol enabling unambiguous identification of each lot
which must be the date and place of sampling and other data that can be
to the analyst.

Fourth
Specific provisions


4.1 Different types of lots

Packing food commodities may have to trade freely
form of bulk foods, food containers, or individual packings
(sacks, bags, retail packings, etc.).
Collection of samples can be applied to all the different forms in which the product is being marketed
.

Without prejudice to the specific provisions of paragraphs 4.3, 4.4 and 4.5 of this Annex
can be used the following formula as a guide for the sampling
batches that are trading form individual packages (bags
bags, retail packaging etc.)
weight of the lot x weight of the incremental sample
sampling frequency = -------------------------------------- -------------------
Weight of the aggregate sample x weight of each package

- Weight: in kg

- Sampling frequency: every nth sack or bag from which they must be

Removed subsample (decimal places are rounded to the nearest whole number
).

4.2 Weight of the incremental sample

Weight of the incremental sample should be 100 g, unless this annex
otherwise specified. For lots in retail packing, the weight depends
subsample to the weight of the retail pack.

4.3 Sampling procedure for cereals, dried vine fruit and roasted coffee

Table 1: Distribution of the consignments, depending on product and lot weight


----------------------------------------------------------------------------------------------------------------------------
Commodity Lot weight Weight Weight neboPočet sub
(T) the number částívzorků general
Batch sample
(Kg)
----------------------------------------------------------------------------------------------------------------------------
Cereals and> = 1500 500t100 10
cereal> 300 and <1,500 3 of 10 šarže100
> = 50 and <= 300 100t100 10
<50 -3 to 1001) 1-10
----------------------------------------------------------------------------------------------------------------------------
Dried grapes> = 15 15 to 10 30t100
(currants, raisins and sultanas
) <15 -10 to 1002) 1-10
----------------------------------------------------------------------------------------------------------------------------
Roasted coffee beans, ground> = 15 15 to 10 30t100
roasted coffee and soluble <15 -10 to 1002), 1-10
coffee
----------------------------------------------------------------------------------------------------------------------------
1) Depending on the lot weight - see Table 2 of this Annex.
2) Depending on the lot weight - see Table 3 of this Annex.
----------------------------------------------------------------------------------------------------------------------------

4.4 The sampling procedure for cereals and cereal products (lots> = 50
t) and for roasted coffee beans, ground roasted coffee, soluble coffee
dried vine fruit (lots> = 15 t) | ||
Sublot may be separated physically, each lot must be physically
divided into parts according to Table 1. Given the weight of the lot
not always an exact multiple of the weight of the batch, the weight of the lot
exceed the mentioned weight by a maximum of 20%.

Each sublot must be sampled separately.

Number of sub-samples is 100.

The aggregate sample is 10 kg.

If it is not possible to use the abovementioned method of sampling
because of the commercial consequences resulting from damage to the lot (because of
packaging forms etc.) can be used
alternative method of sampling provided that it is as representative as possible and is fully described and documented
.

4.5 Guidelines for sampling cereals and cereal products (lots <50 tonnes)
and roasted coffee beans, ground roasted coffee, soluble coffee, dried
vine fruit (lots <15 tonnes)

For cereal lots under 50 the roasted coffee beans, ground roasted
coffee, soluble coffee and dried vine fruit under 15 tonnes may be
depending on the weight of the lot used sampling plan consisting of 10 || | to 100 incremental samples, resulting in an aggregate sample of 1 to 10
kg. For very small lots (ú 0.5 tons) of cereals and cereal products can
remove the lower number of samples, but the aggregate sample uniting
sub-samples must be at least 1 kg.

The numbers shown in the table can be used to determine the number of incremental
samples to be taken.
Table 2: Number of incremental samples to be taken
depending on the weight of the lot



+ ---------------------------------- + ----------- ------------------- +
| Lot weight (tonnes) | Number of incremental samples |
+ ---------------------------------- + ----------- ------------------- +
| = <0.05 | 3 |
| > = 0.05 <0.5 | 5 |
| > 0.5 = <1 | 10 |
| > 1 = <3 | 20 |
| > = 3 to <10 | 40 |
| > = 10 and <20 | 60 |
| = 20 to <50 | 100 |

+ ---------------------------------- + ----------- ------------------- +
Table 3: Number of incremental samples to be taken
depending on the weight of the lot of roasted coffee beans, ground || | roasted coffee, soluble coffee and dried vine fruit


+ ---------------------------------- + ----------- ------------------- +
| Lot weight (tonnes) | Number of incremental samples |
+ ---------------------------------- + ----------- ------------------- +
| <= 0,1 | 10 |
| > = 0.1 <0.2 | 15 |
| > = 0.2 and <0.5 | 20 |
| > = 0.5 <1.0 | 30 |
| > = 1.0 to <2.0 | 40 |
| > = 2.0 to <5.0 | 60 |
| > = 5.0 to <10.0 | 80 |
| > = 10.0 to <15.0 | 100 |
+ ---------------------------------- + ----------- ------------------- +

4.6 Sampling procedure for foods intended for infants and young children ^ 29)

Apply the sampling procedure for cereals and cereal
in paragraph 4.5 of this Annex. Number of incremental samples to be
taken depends on the weight of the lot. According to Table 2 in Section 4.5 of this Annex
a minimum of 10 and maximum of 100 sub-samples.

Weight of the incremental sample should be 100 grams. For lots in retail packing
depends on the weight of the incremental sample weight
retail packing.

The aggregate sample has to be 1-10 kg;
sample must be sufficiently mixed.

4.7 Sampling at retail

Sampling of foodstuffs at retail stage shall be done where possible
accordance with the above sampling provisions. If this is not possible, you can
use other effective sampling procedures at retail
if the sampled lot sufficiently representative.

4.8 Guidelines for sampling wine and grape juice

Aggregate sample shall be at least 1 kg except where
this is not possible, for example, if the sample consists of one bottle.

Minimum number of incremental samples to be taken from the lot is
shown in Table 4. The number of designated sub-samples depends on the form in which
products concerned are usually marketed. For a free
bulk liquid products the lot immediately prior to sampling
manually or mechanically thoroughly mixed, while not affect
product quality. From the lot shall be taken at least three sub-samples
form the aggregate sample.

Sub-samples which are most frequently the form of bottles or packaging must have
same weight. Weight of the incremental sample should be at least 100 g
such that uniting the incremental samples originated aggregate sample
least 1 kg. Deviation from this procedure must be recorded in
sampling protocol.
Table 4: Minimum number of incremental samples to be taken from the lot


+--------------------------------+-----------------------------------+----------------------------+
| Form of marketing | Lot weight expressed | Minimum number of incremental |
| | in volume units (l) | samples to be |
| | | taken |
+--------------------------------+-----------------------------------+----------------------------+
| Bulk products (grape | - | 3 |
| juice, wine) | | |
| Bottles / grape juice packs | = <50 | 3 |
| Bottles / grape juice packs | > 50-500 | 5 |
| Bottles / grape juice packs | > 500 | 10 |
| Bottles / wine packing | = <50 | 1 |
| Bottles / wine packing | > 50-500 | 2 |
| Bottles / wine packing | > 500 | 3 |
+--------------------------------+-----------------------------------+----------------------------+

Fifth

Acceptance of a lot or sublot

Lot or sublot is accepted if the aggregate sample conforms to the maximum limit
taking into account the measurement uncertainty and correction for
recovery.

Lot or sublot is rejected if the aggregate sample
taking into account the measurement uncertainty and correction for recovery
exceeds the maximum limit.


Příl.2
Methods of sampling for the official control of dioxins

(Dibenzo-1,4-dioxin / dibenzofuran) and the determination of polychlorinated biphenyls
dioxin-like PCBs in certain foodstuffs

First

Purpose and scope

Samples intended for the official control of dioxins (1,4
dibenzo-dioxins / dibenzofurans) and also for the determination of polychlorinated biphenyls
dioxin-like PCBs in foodstuffs shall be taken according to the methods described below
. Aggregate samples thus obtained are considered
representative of the lots or sublots from which they were removed.
Compliance with maximum limits laid down in the Regulation of the European Communities
no. 466/2001, setting maximum levels for certain contaminants
in food are determined based on the amount
determined in the laboratory samples.

Second


Definition
Lot: an identifiable quantity of food delivered at one time
which holds an official determination
uniform characteristics such as origin, variety, type of packing, packer, consignor or markings
. For fish and fishery products must be comparable also
size fish.

Part of the batch: a certain part of a large lot allocated to it was
sampled. Each sublot must be physically separate and identifiable
.

Incremental sample: a quantity of material taken from a single place in the lot or sublot
.

Aggregate sample: the combined total of all the incremental samples taken from the lot or sublot
.

Laboratory sample: a representative part / quantity of the aggregate sample intended for laboratory
.
Table toxic equivalency factors (TEF) for risk assessment
for humans (based on the conclusions of the World Health Organization
in Stockholm, Sweden, on June 15 to 18
1997) *) | ||
----------------------------------------------- ----------------------
congener congener Value Value
TEF TEF
--------------------------------------------- ------------------------
Dibenzo-1,4-dioxins (PCDDs), polychlorinated
biphenyls,
polychlorinated
biphenyls without carbon
chlorine in the ortho
positions and
polychlorinated
biphenyls one
chlorine atom
in the ortho position with
dioxin
--------------------------------------------- ------------------------ 2,3,7,8-
1
polychlorinated biphenyls Tetrachlordibenzodioxin without carbon
chlorine in the ortho

positions ---------------------------------------------- ----------------------- 1
1,2,3,7,8- polychlorinated biphenyls 0.0001
Pentachlordibenzodioxin 77
-------------------------------------------------- -------------------
1,2,3,4,7,8- 0.1 polychlorinated biphenyls 0.0001
Hexachlordibenzodioxin 81
-------------------------------------------------- -------------------
1,2,3,6,7,8- 0.1 0.1 polychlorinated biphenyls 126 Hexachlordibenzodioxin

-------------------------------------------------- -------------------
1,2,3,7,8,9- 0.1 polychlorinated biphenyls 0.01 Hexachlordibenzodioxin 169

-------------------------------------------------- ------------------- 1,2,3,4,6,7,8- 0.01


---- Heptachlordibenzodioxin -------------------------------------------------- ---------------

Oktachlordibenzodioxin 0.0001 ------------------------- --------------------------------------------
dibenzofurans (PCDF ) polychlorinated
biphenyls one
chlorine atom
ortho -------------------------------------------
--------------------------
2,3,7,8- 0.1 polychlorinated biphenyls 0.0001
tetrachlorodibenzofurans 105
----------------------------------------------- ----------------------
1,2,3,7,8- 0.05 0.0005 polychlorinated biphenyls
Pentachlordibenzofuran 114 || | ------------------------------------------------- --------------------
2,3,4,7,8- 0.5 polychlorinated biphenyls 0.0001
Pentachlordibenzofuran 118
- -------------------------------------------------- ------------------
1,2,3,4,7,8- 0.1 polychlorinated biphenyls 0.0001
Hexachlordibenzofuran 123
- -------------------------------------------------- ------------------
1,2,3,6,7,8- 0.1 polychlorinated 0.0005


Hexachlordibenzofuran biphenyls 156 ----------------------------------------- ----------------------------
1,2,3,7,8,9- 0.1 polychlorinated 0.0005

Hexachlordibenzofuran biphenyls 157 ----------------------------------------- ----------------------------
2,3,4,6,7,8- 0.1 polychlorinated 0.00001

Hexachlordibenzofuran biphenyls 167 ----------------------------------------- ----------------------------
1,2,3,4,6,7,8- 0.01 polychlorinated 0 , 0001

Heptachlordibenzofuran biphenyls 189 --------------------------------------- ------------------------------
0.01 1,2,3,4,7,8,9- -------------------------------------------

Heptachlordibenzofuran --------------------------

Oktachlordibenzofuran 0.0001 -------------- -------------------------------------------------- -----
*) source - Van den Berg et al. (1998) Toxic Equivalency Factors
(TEFs) for PCBs, PCDDs, PCDFs for Humans and for Wildlife.
Environmental Health Perspectives, 106 (12), 775.

Third


General Provisions 3.1
Staff

Sampling must be performed by an authorized officer (§ 3 para. 1 of this Decree
).

3.2 Material to be removed

Each lot which is to be examined must be sampled separately.

1.3 Precautions

Of sampling and preparation of the samples must be taken
precautions to avoid any changes that
would affect the content of dioxins and polychlorinated biphenyls with
like PCBs, adversely affect the analytical determination or | || aggregate samples unrepresentative.
Sub-samples 3.4


Incremental samples shall, if possible, from various places throughout the lot or sublot
. Deviations from this procedure must be recorded in the log
under 3.8.

3.5 Preparation of aggregate sample

Aggregate sample is made up by uniting all incremental samples. It should have
weight 1 kg, if possible, e.g. if performed taking one
package.

3.6 Distribution of aggregate sample in laboratory samples for confirmation
defense in a trade dispute or arbitration testing

Laboratory samples to confirm, defense or
dispute for arbitration shall be taken from the homogenised aggregate sample
if it does not conflict with § 3 and 4 hereof.
Size of the laboratory samples for confirmation should be sufficient for at least
duplicate tests.

3.7 Packaging and transmission of aggregate and laboratory samples

Each aggregate and laboratory sample shall be placed in a clean container of
inert material that provides adequate protection from contamination
, loss of analytes by adsorption to the internal walls of the vessel and
against damage during shipment. They must be taken all the necessary
preventive measures to avoid change of composition of the aggregate and
laboratory samples, which might arise during transportation or storage.

3.8 Sealing and labeling of aggregate and laboratory samples

Each sample taken for official use shall be sealed at the place of sampling and identified following
§ 6 of this Decree. For each sampling point must be exposed
protocol, which allows unambiguous identification of the lot and the
which must be the date and place of sampling and other data that can be
to the analyst.

Fourth
Sampling plans


The sampling method applied shall ensure that the aggregate sample is representative for
controlled batch.

04.01 Number of subsamples

Case of milk and oils, which are expected steady expansion of
contaminant in the batch, just take three incremental samples per lot,
form the aggregate sample. Give the batch number.
For other products, the minimum number of primary samples to be taken from
batch, given in Table 1.

The aggregate sample uniting all incremental
samples must be at least 1 kg (see section 3.5). Incremental samples must be
similar weight.

Weight of the incremental sample should be at least 100 grams.
Weight of the incremental sample depends on the size of the particles in the lot. Deviations from this procedure
must be recorded in the record provided for under 3.8. In accordance with the provisions
Commission Directive 97/747 / EC of 27 October 1997 laying down
scope and frequency of sampling in accordance with Council Directive 96/23 / EC

Monitoring of certain substances and residues in animal products,
is a sample of chicken eggs at least 12 eggs (for both batch nabalených
eggs and lots consisting of individual packages, Tables 1 and 2
) .

Table 1 Minimum number of incremental samples to be taken from the lot

----------------------------------------------- -------------------
lot weight (kg) Minimum number of samples of sub
Sample
---------------------------------------------- --------------------
<50 3
50-500 5
> 500 -------- 10
-------------------------------------------------- --------

Consists if a batch of individual packages, the number of packages that must be removed
to create the aggregate sample is given in Table 2. Table 2

Number of packages (incremental samples, which must be removed to create
aggregate sample consisting If lots of individual packs
)

-----------------------------------------------
------------------- Number of packages or units Number of packages or taken

units in the lot ----------- -------------------------------------------------- -----
1 to 25 1 package or unit
-------------------------------- ----------------------------------
26-100 about 5%, at least two packages or || |
units ---------------------------------------------- --------------------
> 100 about 5%, maximum 10 packages or

units ---------------------------------------------- --------------------

4.2 Specific provisions for the sampling of lots of whole fish

The number of incremental samples to be taken from the lot are defined in Table
first The aggregate sample uniting all incremental
samples must be at least 1 kg (see section 3.5).

If the lot to be sampled contains also individual fish weighing less than 1 kg
, it is taken for bulk sample as a whole fish subsample.
If the weight of the thus formed aggregate sample greater than 3 kg
subsample may consist of the middle part of the fish weighing at least 100 g
that the aggregate sample. The whole part, covered
maximum limit is used for homogenisation of the sample.

If the lot to be sampled contains individual fish weighing more than 1
kg, an incremental sample the middle part of the fish. Each partial sample has a weight of at least 100
g. If the sampled lot consists of fish weighing more than 6 kg
and removing the central part would be of considerable economic
loss taken at least three samples of a minimum of 350 grams without
regardless of batch size.

Fifth

Specification Compliance of the lot or sublot

The lot is accepted if the analytical result does not exceed the maximum
limit laid down in Regulation (EC) no. 466/2001, taking into account the measurement uncertainty
.

The lot is rejected if the analytical result confirmed by duplicate
test sample and calculated as the mean of at least two separate determinations exceeds
doubt taking into account the measurement uncertainty
maximum limit laid down in Regulation (EC) no. 466/2001 .

Measurement uncertainty may be taken into account by one of the following ways:

A) calculating the expanded uncertainty, using a coverage factor of 2, which corresponds
confidence interval of approximately 95%, or

B) establishing the decision limit (CCalfa)
by Commission Decision 2002/657 / EC of 12 August 2002 implementing Council Directive
96/23 / EC concerning the performance of analytical methods and
interpretation of results.


Příl.3
Sampling procedure for checking compliance with maximum levels
lead, cadmium, mercury and 3-chloro-propane-1,2-diol for certain foodstuffs and raw materials


First

Purpose and scope

Samples intended for official control of the levels of lead, cadmium, mercury and
3-chloro-propane-1,2-diol in foodstuffs shall be taken following
methods. Aggregate samples thus obtained shall be considered
representative for the lot or part of the batch from which they were removed.
Compliance with maximum limits laid down in the Regulation of the European Communities
no. 466/2001 will be determined on the basis of the levels determined in the laboratory samples
.

Second


Definition Batch
:
identifiable quantity of food delivered at one time, which

Holds an official determination of the same characteristics, such as origin,
variety, type of packing, packer, consignor or markings. Fish must also have
comparable size.
Part of the batch
:
fixed part of a large lot allocated to it was made by sampling
. Each sublot must be physically separate and identifiable
.

Incremental sample:
quantity of material taken from a single place in the lot or sublot.

Aggregate sample:
summary of all the incremental samples taken from the lot or sublot.

Laboratory sample: sample intended for laboratory examination.

Third
General

3.1
Staff

Sampling must be performed by an authorized qualified personnel (§
paragraph 3. 1 hereto).

3.2 Material to be removed

Each lot which is to be examined must be sampled separately.

3.3 Precautions

Of sampling and preparation of the samples must be taken
precautions to avoid any changes, which would affect
contents of lead, cadmium, mercury and 3-chlorpropan-
1,2- diol, adversely affect the analytical determination or degrade
representativeness aggregate samples.
Sub-samples 3.4


Incremental samples shall, if possible, from various places throughout the lot or sublot
. Deviations from this procedure must be recorded in the log
under 3.8.

3.5 Preparation of aggregate sample

Aggregate sample is made up by uniting all incremental samples.
Should weigh at least 1 kg unless it is impossible, for example.
When sampling a single package.

3.6 Distribution of aggregate sample in laboratory samples for examination
repeated investigations and examinations referee

Laboratory samples for examination, repeated testing and the referee
shall be taken from the homogenised aggregate sample unless this
not conflict with § 3 and 4 hereof.
Size laboratory sample for testing shall be sufficient for at least two tests.

3.7 Packaging and transport of aggregate and laboratory samples

Each aggregate and laboratory sample shall be placed in a clean container of
inert material that provides adequate protection from contamination
, analyte loss due to adsorption on the inner walls
container and against damage in transit. They must be taken
all necessary measures to avoid any change in composition of the aggregate and
laboratory samples, which might arise during transportation or storage.

3.8 Sealing and labeling of aggregate and laboratory samples

Each sample taken for official use shall be sealed at the place of sampling and identified following
§ 6 of this Decree. For each sampling point must be exposed
protocol allowing clear identification of the sample, which
shall be the date and place of sampling and other data that can be
to the analyst.

Fourth

Sampling plan

Ideally, it should be sampled at the point where
commodity enters the food chain and where we can distinguish individual
lot. The sampling method applied shall ensure that the aggregate sample is representative for
controlled batch.

4.1 Number of subsamples

For liquid products, which are expected
uniform extension of the contaminant in the batch, simply remove one subsample of
batch, which will aggregate sample. Give the batch number.
Liquid products containing hydrolysed vegetable protein or soy sauce
fluid must before fractional sampling shaken well, or homogenised
other appropriate manner.

The other products are removed from the lot Minimum number of primary samples
listed in Table 1. The incremental samples shall be of the same weight.
Deviations from this procedure must be recorded in accordance with point
08.03
Table 1:
Minimum number of incremental samples to be taken from the lot

----------------------------------------------- ---------------
lot weight (kg) Minimum number of samples
subsamples
--------------------------------------------- -----------------
<50 3
50-500
5> 500 10
----------- -------------------------------------------------- -

In addition, when a batch of individual packages, it is taken for bulk sample number

Samples listed in Table 2. Table 2
:
Number of packages (incremental samples) to be removed for the general
sample it forms a batch of individual packages
----- --------------------------------------------------
----------- Number of packages or units Number of packages or taken

units in the lot ------------------- -----------------------------------------------
1 to 25 1 package or unit

26-100 about 5%, at least two packages or
unit

> 100 about 5%, maximum 10 packages or

units ---------------------------------------------- --------------------

Fifth

Compliance with the highest content specified in the lot or sublot

For control purposes, performs control laboratory
least two independent tests and the results averaged.

The lot is accepted, if the average of the highest relevant content
laid down in Commission Regulation (EC) no. 466/2001, taking into account the
expanded measurement uncertainty and correction for recovery following the
European Commission report on the relationship between analytical results, measurement
uncertainty, recovery factors and EC legislation in the field
food.

The lot is rejected if it exceeds the maximum diameter of the respective content
taking into account the expanded measurement uncertainty and correction for
recovery.
Appendix 4



Canceled Annex 5


Preparation procedures and criteria for the test methods used in determining
levels of ochratoxin A in certain foodstuffs and raw materials

First Preventive measures

Because the distribution of ochratoxin A is very inhomogeneous, should
samples shall be prepared, and especially homogenised, with extreme care.

All the material received by the laboratory is to be used for the preparation of test material
.

Second Treatment of the sample received in the laboratory

Each laboratory sample finely grinded and mixed thoroughly process
which achieves complete homogenisation.

If the maximum level applies to the solids content is determined in part
homogenised sample, using a method that has been demonstrated to
accurate determination of dry matter content. ".

Third Distribution of samples for examination under to confirm and defense trade dispute


Repeated samples for testing to confirm, defense
business and referee purposes shall be taken from the homogenised material
if it does not conflict with § 3 and 4 of this Decree
.

4. methods for testing in the laboratory use, and requirements for management of laboratory



4.1 Definitions the following are some of the most common definitions that the laboratory used
.
the most commonly quoted precision parameters are repeatability and reproducibility

r = repeatability - the value below which will be as expected with the
probability (typically 95%) lie or it will be equal
absolute difference between single test results
under repeatability conditions (ie. same sample, same
operator, same apparatus, same laboratory, established shortly
consecutive);

R = 2.8 x sr

Sr = standard deviation - calculated from results
Repeatability

RSDr = relative standard deviation - calculated from results
obtained under repeatability conditions [(sr / x) x 100], where
x is the average of results over all laboratories and samples

R = reproducibility - the value below which the expected
with a certain probability (typically 95%), or it will lie
equal to the absolute value of the difference between the results of two separate
under reproducibility conditions (ie. the same
material obtained by operators in different laboratories, using the standardized test method
); R = 2.8 x SR

Standard deviation - calculated from results under conditions
reproducibility

RSDR = relative standard deviation - calculated from results
generated under reproducibility conditions [(SR / x) x 100];

4.2 General requirements Testing methods used for control purposes
food should be, whenever possible, in accordance with § 9 of this decree.
4.3 Specific requirements
If not at the level of the European Communities
prescribed specific methods for the determination of ochratoxin A in foods
, laboratories may select any method provided

That meets the following criteria:
Characteristics of the efficacy of the method for the determination of ochratoxin A

----------------------------------------------- ------------------

levels of ochratoxin A microg / kg
-----------------------------------------------
RSDr (%) RSDR (%) Recovery (%) -----------------------------------
------------------------------
<1 = <40 = <60 50-120 ---
-------------------------------------------------- ------------
1-10 = <= 20 <30 70 to 110 --------------------
---------------------------------------------

- Detection limits of the methods used are not stated as the precision
It is given at the concentrations.
- The precision values ​​are calculated from the Horwitz equation:

RSDR = 2 E (1-0.5 log C)
Where
:
- RSDR is the relative standard deviation calculated from results
generated under reproducibility conditions [(sR / x) x 100]
- c is the concentration ratio (i.e. 1 = 100 g / 100 g, 0.001 = 1000
mg / kg).

This is a generalized equation for the accuracy with which it is shown that for most
routine testing methods independent of analyte and matrix but only
concentration.

4.4 Recovery calculation and reporting of results

The test results are reported corrected or uncorrected for recovery.
Must The manner of yield and its value.
Test result corrected for recovery is used for checking compliance
(see Annex no. 41 point 5).

The test result must be presented in the form (x +/- U), where x is
analytical result and U is the expanded measurement uncertainty.

U is the expanded uncertainty, using a coverage factor of 2
which gives a level of confidence of approximately 95%.

4.5 Standards Laboratory quality

Laboratories must comply with the special regulation. ^ *)
Appendix 6


Sample preparation and criteria for test methods for checking compliance
levels of lead, cadmium, mercury and 3-chloro-propane-1,2-diol
certain foodstuffs and raw materials

First

Introduction

Basic requirement is to obtain a representative and homogeneous laboratory sample
, with no secondary contamination.

Second Specific procedures for preparing samples for lead, cadmium and mercury

There are a number of compliant sample preparation procedures which may be used
for the products concerned. The procedures outlined in the draft European standard
"Foodstuffs. Determination of trace elements. Requirements for efficiency and
general principles." They have proved satisfactory, but may be equally
other procedures.

For each procedure must be complied with these rules:

- Mussels, crustaceans and small fish: if eaten whole, the viscera must be
included in the material to be tested

- Vegetables: investigates only the edible portion of which must be taken into account
requirements of the European Communities no. 466/2001.

Third

Testing methods, used by the laboratory and laboratory requirements for testing

3.1 Definitions


Below are a few common definitions that the laboratory used
:
r = repeatability - the value below which will be as expected with the
probability (typically 95%) or it will be equal to the absolute | || the difference between single test results under conditions
repeatability (ie. the same sample, same operator, same
apparatus, same laboratory, established in quick succession);

R = 2.8 x sr;

Sr = standard deviation - calculated from results
repeatability conditions;
RSDr = relative standard deviation - calculated from results
obtained under repeatability conditions [(sr /) x 100], where
the average of results over all laboratories and samples;

R = reproducibility - the value below which the expected
with a certain probability (typically 95%), or it will lie
equal to the absolute value of the difference between the results of two separate
under reproducibility conditions (ie. the same
material obtained by operators in different laboratories, the same procedure
); and therefore

R = 2.8 x SR;

Standard deviation - calculated from results under conditions
reproducibility;

RSDR = relative standard deviation - calculated from results
generated under reproducibility conditions [(SR / x) x 100];

HORRATr = value found RSDr divided by the RSDr value calculated
from the Horwitz equation using the assumption r = 0,66R;


HORRATR = RSDR value divided by the RSDR value calculated
from the Horwitz equation.
3.2 General requirements


Testing methods used for food control purposes must comply whenever
possible, in accordance with § 9 of this decree.

For the analysis of lead in wine is prescribed method given in
Chapter 35 of the Annex to Regulation Commission of the European Communities no. 2676/90
establishing methods for testing wines.
3.3 Specific requirements


3.3.1 Testing of lead, cadmium and mercury

Specific methods for the determination of lead, cadmium and mercury contents are not prescribed
. Laboratories use validated methods that meet
efficiency characteristics shown in Table 3. The test materials
in the collaborative test laboratory for the validation of methods should
feasible, include a certified reference material.
Table 3: Characteristics of the efficiency of methods for testing the contents
lead, cadmium and copper

Characteristic Value / comment

Applicability Food specified in Regulation of the European
Communities no. 466/2001

The limit of detection may not be greater than one tenth of the value
specified in the Regulation of the European Communities
no. 466/2001, except when it is for
Lead stated value of less than 0.1
mg / kg. In this case, should not exceed
fifth the values ​​

Limit shall not be higher than one-fifth of the value
specified in the Regulation of the European Communities
no. 466/2001, except when it is for
Lead stated value of less than 0.1
mg / kg. In this case, should not exceed
two-fifths of the values ​​

Accuracy values ​​HORRATr or HORRATR of validation
ring test must be less than 1.5

Yield 80 to 120% (according to the ring
validation test)

Specificity must not disturb the matrix or other substances at
Spectral analysis

3.3.2 Testing of 3-chloropropane-1,2-diol

Specific methods for the determination of 3-chloropropane-1,2-diol are
prescribed. Laboratories use validated methods that meet
performance characteristics listed in Table 4. Test
materials used in the ring test laboratories for the purpose of validation of the method should contain
possibly certified reference material.
Specific method has been validated in a ring test and fulfilled the requirements set out in Table 4.

Table 4: Characteristics of the effectiveness of methods for testing
3-chloro-propane-1,2-diol

----------------------------------------------- ------------------
Characteristics Recommended value Concentration
---------------------- -------------------------------------------
blind test lower than The limit of detection -
-------------------------------------------- ---------------------
yield of 75-110% full scope

concentration ---------------------------------------------- -------------------
limit of quantification 10 (or less) -
microg / kg based on

dry matter ---------------------------------------------- -------------------
The standard deviation of less than 4 microg / kg -

blinded ----------- -------------------------------------------------- ----
Estimates precision within <4 microg / kg, 20 microg / kg
laboratories - standard <6 microg / kg 30 mcg / kg
deviation of repeated <7 mcg / kg of 40 microg / kg | || measurements at different <8 mcg / kg of 50 microg / kg
concentrations <15 microg / kg, 100 microg / kg
-------------------- ---------------------------------------------

3.3.3 Operational characteristics - the concept of uncertainty
suitability test methods to be used in the laboratory
can be assessed also using the concept of uncertainty.
The laboratory may use a method which will produce results within a maximum standard uncertainty
. The maximum standard uncertainty is calculated using the equation
:
+ +
| 2 2 |
Phew root = | (LOD / 2) + (Alfac) |
+ +
Where:
Uf is the maximum standard uncertainty
LOD is the limit of detection methods
C is the concentration,
alpha is a numerical factor used depending on the value of C.
The values ​​to be used are given in Table
no. 5:


Table 5: Values ​​of numerical factor alpha based on
worth C
-----------------------------------------------
C (microg / kg) alpha
-----------------------------------------------
= <50 0.2
-----------------------------------------------
51-500 0.18
-----------------------------------------------
501-1 000 0.15
-----------------------------------------------
1001 - 10000 0.12
-----------------------------------------------
> = 10 000 1
-----------------------------------------------

Uf is the expanded measurement uncertainty using a coverage factor
2, which provides a confidence level of approximately 95%.

If a test method provides results with uncertainty
Measuring less than the maximum standard uncertainty, the method will
Suitable as well as a method that meets labor
characteristics listed in Tables 3 and 4.

3.4 Estimation of the accuracy tests, recovery calculation and reporting of results

Whenever possible, the Company estimates the accuracy of test that is performed
control test suitable certified reference material.

Test results will be reported as corrected or uncorrected for
recovery. This information must be included in the test report
well as yield.

Be taken into consideration to a European Commission report on the relationship between analytical results,
measurement uncertainty, recovery factors and EC legislation in the field
food.

The test result must be reported as x +/- U, where x is the result
and U is the measurement uncertainty.

3.5 Laboratory quality standards

Laboratories must comply with the provisions of special legislation. ^ *)


3.6 Expression of results
The results are expressed in the same units, in which are set
maximum levels in the European Community regulations no. 466/2001.


Příl.7
Sample preparation and requirements for the test methods used for the determination of levels of dioxins
(dibenzo-1,4-dioxin) and determination of polychlorinated biphenyls
dioxin-like PCBs in certain foodstuffs

First

Objectives and scope

These regulations apply to testing for the official control of foodstuffs
levels of dioxins (polychlorinated dibenzo-1,4-dioxin) and
polychlorinated dibenzofurans and polychlorinated biphenyls determination
with dioxin.

Monitoring the presence of dioxins in foodstuffs can be based on
strategy using a screening method to find samples with levels of dioxins
about 30-40% lower or higher than the interest level.
The concentration of dioxins in those samples with significant numbers
to establish or confirm a confirmatory method.

Screening methods are serving to detect dioxins and polychlorinated biphenyls
dioxin-like PCBs at the level of interest.
These methods have a high capacity in terms of number of samples, and are used to sift
potentially positive samples from a large number of samples
. They are specifically designed to avoid false negative results
.

Confirmatory methods are methods that provide full or complementary information enabling
clearly qualitatively and quantitatively determine
dioxins on the interest level.

Second
Basic information


Given that the samples of the environment carried
and biological samples (including samples of foodstuffs) usually contain complex mixtures of different dioxin congeners
was to facilitate risk assessment
introduced the concept of toxic equivalency factors. Factors toxic equivalence
been designed to reflect the concentration of a mixture
substituted 2,3,7,8-dibenzo-1,4-dioxins and polychlorinated dibenzofurans
, and more recently some polychlorinated biphenyls
without chlorine atoms in the ortho positions or one chlorine atom in the ortho-position
which exhibit dioxin-like activity in toxic
equivalents (TEQ) of 2,3,7,8-tetrachlordibezodioxinu.
Concentrations of the individual substances in a given sample are multiplied by their respective toxic equivalency factors
, added together and the resulting sum is the total
concentrations of dioxin-like compounds expressed in toxic equivalent
.

In the method of "upper-bound" is the size of the contribution of undetermined
quantified congener to the toxic equivalent equal to the value of the limit of quantification
.


In the method of "lower bound" is the size of the contribution of undetermined
quantified congener to the toxic equivalent of zero. In the method
"middle estimate" is the size of the quantitative contribution of undetermined
congener to the toxic equivalent equal to half of the limit of quantification
.

For the purposes of this Annex
accepted specific limit of quantification of an individual congener is the concentration of an analyte in a sample extract
that a measuring instrument provides for the two different ions that
to be monitored, the response ratio signal / noise (S / N) of 3: 1 for at least
sensitive signal and fulfills the requirements according to methods of determining
described in EPA method 1613 Revision B.

Third

Requirements for quality assurance, which must comply with sample preparation

- At each stage of the sampling and testing must be taken to avoid cross-contamination
.

- The sample must be stored and transported in glass, aluminum, polypropylene or polyethylene
. Of the sample container must be removed
Traces of paper dust. The glass is rinsed with solvents which have been previously
checked for the presence of dioxins.

- The sample must be transported and stored so as to maintain the integrity
food sample.

- If necessary, every laboratory sample is ground finely and mixed thoroughly
process that is shown that to achieve complete
homogenization (e.g. by milling and sieving through a 1 mm sieve); if
sample moisture is too high, the samples are dried before grinding.

- Carry out a blank test without the sample using
entire analytical process.

- Weight of sample used for extraction must be sufficient to be
meet the requirements of sensitivity.

- There are many satisfactory sample preparation procedures which may be relevant for
samples used. Procedures must be validated by
internationally recognized methodologies.

Fourth

Requirements for laboratories

- Laboratories shall demonstrate the functionality of the method in the range of interest
levels, e.g. at the level of interest while, at its middle or its
twice, with an acceptable coefficient of variation for repeated
test. For details of acceptance criteria are listed in note 5.

- The limit of determination for confirmatory method should be at the level of one fifth
interest levels to ensure that the level of interest will
achieve acceptable coefficients of variation.

- As a measure of internal quality control should be performed regularly
blind experiments, experiments with artificially fortified blank samples or tests
control samples (preferably certified reference material
).

- Successful participation in interlaboratory comparison tests in which the laboratory
assesses proficiency is the best way to verify the technical competence
for specific tests. Successful participation in interlaboratory tests
eg. For samples of soil or sludge, is not necessarily proof of professional competence
in the food or feed in which there are lower levels of contamination
. Therefore, it is mandatory continuous participation in interlaboratory tests
determination of dioxins and dioxin-like polychlorinated biphenyls
PCBs in food matrices corresponding feed.

- In accordance with § 9 of this decree should be
laboratories accredited by a designated authority operating in accordance with the guidelines of the International Standardization Organization
no. 58 to ensure that they are applying
system of quality assurance. Laboratories should be accredited according to
ISO / IEC / 17025: 1999.

Fifth

Requirements to be met by analytical method for the determination of dioxins and polychlorinated biphenyls
dioxin

Basic requirements for acceptance of analytical procedures:

- High sensitivity and low detection limit. In the case of dibenzo-1,4-dioxin and polychlorinated dibenzofurans
must be because of extreme toxicity
some of these compounds can be detected at picogram quantities
level toxic equivalency (10-12 g). It is known that polychlorinated biphenyls
occur at higher concentrations than 1,4-dibenzo-dioxins and polychlorinated dibenzofurans
. For most polychlorinated biphenyls
is sufficient already nanogram sensitivity level (10-9 g).
To determine more toxic polychlorinated biphenyls with
dioxin (especially unsubstituted congeners with chlorine in

Ortho-positions) must be achieved equally sensitive for determining
dibenzo-1,4-dioxins and polychlorinated dibenzofurans.

- High specificity. Dibenzo-1,4-dioxin, polychlorinated dibenzofurans
and dioxin-like PCBs must
distinguished from other compounds that were extracted together with these
substances may interfere with the determination and are present in | || concentrations up to several orders of magnitude higher than the concentration of interest
analytes. In methods based on gas chromatography with mass spectrometric detection
is necessary to distinguish between the different congeners
i.e. between toxic (eg. seventeen dibenzo-1,4-dioxin and polychlorinated dibenzofurans
substituted 2,3,7,8
PCBs and dioxin-like PCBs) and other congeners.
Bioassays should make it possible to determine the values ​​of toxic equivalent sum for selectively
dibenzo-1,4-dioxins, polychlorinated dibenzofurans and
dioxin-like PCBs.

- High accuracy (trueness and precision).
Determination should provide a valid estimate of the true concentration in a sample.
High accuracy (accuracy of the measurement: the closeness of agreement between a test result
measurement and the true value or agreed value) is necessary to
order was rejected by the test result of the sample based on the unreliability of the estimate
toxic equivalent. Accuracy is expressed
trueness (difference between the mean value obtained by measuring an analyte in a certified material
and certified value expressed as a percentage of this
certified value) and precision (precision is usually
calculated as a standard deviation including repeatability
and reproducibility, and indicates the closeness of agreement between the results obtained by several
repeating the experimental procedure
under prescribed conditions).

Screening methods can be bioassays and methods based on
gas chromatography with detection by mass spectrometry
. Confirmatory methods are methods based on gas chromatography
high-resolution mass spectrometry detection
high resolution. Determining the value of total toxic equivalent
must meet the following criteria:
---------------------------------- -------------------------------
Screening methods Confirmatory methods -------------------------------------------
----------------------
proportion of false <1% - negative results

----------- -------------------------------------------------- ----
Truthfulness - -20% to + 20% -------------------------------
----------------------------------
The coefficient of variation <30% <15%
- --------------------------------------------------
-------------
6th

Specific requirements to be met by a method based on gas chromatography
detection for mass spectrometry for screening and confirmation


- In order to validate the test procedure shall be at the very beginning of the process, such
. before extraction, internal standards added
2,3,7,8-tetrachloro dibenzo-substituted 1,4-dioxins and polychlorinated dibenzofurans
labeled with an isotope 13C (a standard
polychlorinated biphenyls with a dioxin labeled with an isotope 13C
when determining polychlorinated biphenyls and dioxin-like PCBs).
Must be added at least one congener for each of the tetra to oktachlor
dibenzo-1,4-dioxin and dibenzofurans (and at least one congener for each of
groups for dioxin-like PCBs in || | determination of polychlorinated biphenyls and dioxin-like PCBs) or to the
least one congener for each detected ion mass spectrometry
monitoring dibenzo-1,4-dioxin and polychlorinated dibenzofurans
polychlorinated biphenyls and dioxin-like PCBs.
Especially in case of confirmatory methods, the advantage of using all 17
internal standards 2,3,7,8- substituted dibenzo-1,4-dioxin and dibenzofurans
labeled with an isotope 13C and all 12 internal standards polychlorinated || | dioxin-like biphenyls labeled with an isotope 13C in the determination
polychlorinated biphenyls and dioxin-like PCBs.
Using appropriate calibration solutions should also be determined relative response
congeners for which they have been added compounds labeled with an isotope 13C.

- For foodstuffs of plant origin and foodstuffs of animal origin

Fat content of less than 10%, the addition of the internal standard before extracting
mandatory. For foodstuffs of animal origin with a fat content higher than 10%
internal standards may be added either before extraction or after extraction
. Suitable method should be validated efficacy
extraction, depending on the time of addition of the internal standards and
according to whether the results apply to a product or the fat content.

- Before testing by gas chromatography with detection by mass spectrometry
must be added one or two standards for determining
yield.

- Control of recovery is necessary. For confirmatory methods should
recoveries of the individual internal standards lie in the range 60% to 120%
. Lower or higher recoveries for individual congeners, in particular
some hepta- and octa-chlordibenzodioxinů and dibenzofurans are
acceptable, provided that their contribution to the value of toxic
equivalents does not exceed 10% of total toxic equivalent
(based only dibenzo-1,4-dioxins and polychlorinated dibenzofurans
). For screening methods should yield
lie in the interval from 30% to 140%.

- Separation of dioxins from interfering chlorinated compounds such as
polychlorinated biphenyls and chlorinated biphenyl ethers should be carried
suitable chromatographic techniques (preferred adsorbents
florisil, alumina or activated charcoal).

- Resolution gas chromatography of isomers shall be sufficient (ratio
peak between 1,2,3,4,7,8-hexachlordibenzofuranem and 1,2,3,6,7,8-
hexachlordibenzofuranem - < 25%).

- Determination should be performed revised EPA Method 1613 / B
or other method with comparable performance characteristics.

- For foodstuffs with a dioxin contamination of about 1 pg
toxic equivalent (according to the World Health Organization gram of fat -
toxic equivalent of just based on dibenzo-1,4-dioxins and polychlorinated dibenzofurans
) would had an estimated difference between the upper and lower
estimated to exceed 20%. For foodstuffs with a low fat content must be
contamination levels of about 1 pg toxic equivalent (according to the World Health Organization
per gram of product)
met the same requirements. For lower contamination levels, e.g. 0.50 pg toxic
equivalent according to the World Health Organization g product
may be the difference between the upper and lower bound within the range of 25-40%.

7th

Screening test methods

7.1 Introduction Screening methods can be used in different approaches to implementing
tests: a pure screening and the quantitative testing.
Screening approach


Response of the sample is compared with a reference sample of interest
level. Samples with a response less than the reference sample
declared negative, those with a higher response are suspected positives. Requirements:

- Blind and reference samples as classified in each test series, which is
extracted and tested at the same time and under the same conditions.
Reference sample must show a clearly elevated response in comparison to a blank
sample.

- In addition, the ranks reference samples at the half and twice
concentration than the interest level to demonstrate the proper implementation
test the extent consistent with the level of interest.

- When testing other matrices, the suitability of the reference sample
(reference samples), preferentially by including samples which were
using methods based on gas chromatography
high-resolution mass spectrometry detection . Determination of the
gas chromatography with mass spectrometric detection shall be demonstrated
toxic equivalent value close to the value in
reference sample or else a blank spiked at this value.

- Given that in bioassays can not use any internal
standards repeatability tests are very important to obtain
information on the standard deviation within one test series.
Coefficient of variation should be less than 30%.

- For bioassays must be specified target compounds, possible interferences, and
maximum permitted level in the blank.
Quantitative testing


Quantitative testing requires serial dilutions of the standard solution
duplicate or triplicate clean up and measuring as well
inclusion of blanks and control yield. Results can be expressed in

Toxic equivalents, based on the fact that compounds that cause
signal correspond to the principle of toxic equivalent. This can be realized by
tetra chlordibenzodioxinů (or standard mixtures
tetrachlordibenzodioxin / tetrachlorodibenzofurans) with
constructing a calibration curve for the calculation of toxic equivalent extract and thus
sample. This is subsequently corrected to the toxic equivalent of blank
(to account for impurities in the solvents and chemicals)
and a recovery (calculated from the toxic equivalent sample intended for
quality control toxic equivalent to the interest level).
It is necessary to note that the reduction in yield may partly be due
matrix effects or differences between the toxic factors
equivalence in the bioassays and the official values ​​of factors
toxic equivalence according to the World Health Organization.

7.2 Requirements for test methods used for screening

- For screening methods may be used based on
gas chromatography with detection by mass spectrometry and bioassay. For
methods based on gas chromatography with mass spectrometric detection
the requirements set out in paragraph 6. Specific requirements for bioassays
cells are given in section 7.3 and the requirements for bioassays with sets are presented in Note
4.7

- It is necessary to state what is the number of false positive and false negative results
in large sets of samples with values ​​lying
above and below the maximum level or emergency level compared with the results obtained
toxic equivalent confirmatory methods testing.
The actual proportion of false-negative results should be less than 1%.
Proportion of false positive results should be sufficiently low to
was use screening preferred.

- Positive results must be confirmed by a confirmatory method
testing based on gas chromatography, high-definition
mass spectrometric detection with high resolution. Furthermore, it must be
confirmatory method based on gas chromatography with mass spectrometric detection
high resolution results confirmed by
samples with a wide range of values ​​of toxic equivalent (approximately 2% to 10%
negative samples). They should be available compliance information
bioassay results and methods based on gas chromatography
High resolution mass spectrometric detection with high resolution
.
7.3


Specific requirements for bioassays on cells

- When performing bioassay it is necessary to use for each test series
reference concentrations of tetra-chlordibenzodioxinů or mixtures
tetrachlordibenzodioxin / tetrachlorodibenzofurans (full curve
dose-response with r2> 0.95). For screening purposes, however, may be the
testing of samples with low values ​​applied curve extended into areas
lows.

- To express the results of bioassays within
constant time period may be used as a reference concentration tetrachlordibenzodioxinů
(about 3 times higher than the limit of quantification) specified in the protocol of quality control.
Alternative could be the relative response of a reference sample in comparison to a calibration curve
tetra chlordibenzodioxinů, since the response of the cells may
depend on many factors.

- For each type of reference material should be recorded and verified graphs
quality control to ensure that the results are
accordance with established guidelines.

- Especially when quantitative calculations must be used to such thinning
sample within the linear portion of the response curve.
Samples above the linear portion of the response curve must be diluted and
try again. It is therefore recommended to be simultaneously tested
least three or more degrees of dilution.

- Standard deviation expressed as a percentage must be at three
set for any degree of dilution of more than 15% between three independent experiments
not be higher than 30%.

- The limit of detection may be set as three times the standard deviation
solvent blank or of the background response. Another possibility is to use
response that is above the background calculated from the calibration
curve of the day (induction factor five times the response
blank solvent). The limit of determination can be determined

Level five to six times the standard deviation of the solvent blank
or background or apply a response that is above the background response
calculated from the calibration curve of the day (inductive
response factor of ten blind sample solvent).
7.4


Specific requirements for kit bioassays

- The sample preparation and the tests must be adhered to manufacturer's instructions.

- Test kits should not be used after the expiry date.

- Should not use materials or components designed for use with a different set
.

- Test kits should be kept under storage
temperatures and should be used at the specified operating temperature.

- The limit of detection of immunoassays is determined as a percentage of the value response
three times the standard deviation of ten repetitive determination
carried to a blank value and the slope obtained by linear regression
.

- To check if the response is within an acceptable range should
during laboratory tests used reference standards.

8th
Reporting of results


If analytical procedure allows should analytical results
contain values ​​for the individual congeners dibenzo-1,4-dioxin and
dibenzofurans and polychlorinated biphenyls, and it should be indicated whether
regards the upper, lower or middle estimate in order to report on the results
specified maximum amount of information and was thus able to interpret
results according to specific requirements. The report should also be given
lipid content of the sample and the method of lipid extraction.

If recoveries are outside the range mentioned in point 6, or if
exceeded the maximum limit, and, upon request, shall be available
recovery for individual internal standards.


Příl.8

Canceled

Příl.9

Canceled

Příl.10

Canceled

Příl.11
Determination of erucic acid in oils and fats intended as
such for human consumption and the fat or oil component of foods to which
oils or fats added

I.

Introduction

First Sample Preparation
1.1 General


Weight of the sample to the laboratory for examination under normal conditions
50 g unless a larger quantity is required.

02.01 Sample preparation for the examination in the laboratory

The sample must be homogenized before the test.

03.01 Storage containers

Sample thus prepared was stored in an airtight and watertight container.

Second reagents

02.01 water

2.1.1. To dissolve and wash the used distilled or demineralized water
equivalent purity.

2.1.2. If not at the mention of dissolving or diluting stated
no other agent, it is a solution or dilution.

02.02 chemicals

Used only chemicals of analytical purity (pa), unless otherwise indicated
.

Third Apparatus

03.01 The list of devices

This list contains only those items with a special purpose and
specifications.

03.02 Analytical Balance

Term analytical balance means a balance with a sensitivity of 0.1 mg or greater.

Fourth Expression of results

04.01 results

The test report shall be the mean value from at least two
determination with satisfactory reproducibility.

04.02 Calculation of the percentage

Unless otherwise specified, the results are expressed in weight
percentage of the total fatty acids in the sample as received by the laboratory

.

04.03 Number of significant figures

Number of significant figures in the result so expressed is designed
precision of the method.

II.
Determination of erucic acid


First Subject and scope of application

The method determines the erucic acid content

- Oils and fats containing cetoleic acid (Z isomer
docosenoic acid which occurs in fish oils), and

- Hydrogenated oils and fats containing E and Z isomers
docosenoic acid.

Second definition

Term erucic acid content means the content of erucic acid
determined by the method.

Third The principle of the method

Methyl esters of the component fatty acids of the oil or fat are separated
argentation thin-layer chromatography at low temperature and
quantitatively determined by gas-liquid chromatography
phases.

Fourth Reagents


04.01 Freshly distilled ether without peroxides

04.02 n-hexane

04.03 Silica gel G thin layer chromatography

04.04 Silica gel, for column chromatography

05.04 Solution Silver nitrate 200 g / l. In water
Dissolve 24 g of silver nitrate and dilute with water to 120 ml.

06.04 A solution of methyl erucate 5 mg / ml.
In a few ml of n-hexane was dissolved 50 mg of methyl erucate and dilute
n-hexane to 10 ml.

07.04 Methyl tetracosanoate internal standard solution
0.25 mg / ml.

In a few ml of n-hexane was dissolved 25 mg of methyl
tetracosanoate (as in section 4.6.) And dilute with n-hexane to 100 ml.

08.04 Developing solvent: toluene: n-hexane 90:10 (v).

09.04 A solution of 2,7-dichloro-0,5 g / l. For
warming and stirring were dissolved 50 mg of 2,7-
dichlorofluoresceinu in 100 ml of 50% aqueous methanol.

Fifth Apparatus

05.01 Equipment for thin layer chromatography, in particular:

5.1.1. Freezing unit, capable of maintaining developing tank and contents at a temperature of -20
st. C to -25 st. C.

5.1.2. Glass plates 200x200 mm.

5.1.3. UV lamp

5.1.4. Glass columns, length about 200 mm internal diameter about 10 mm
with filter of glass wool or sintered, or small fritted funnel.

5.1.5. Applicator, for depositing solutions of a narrow band or strip to
chromatographic (TLC) plate.

05.02 Gas chromatograph-liquid phase
with an electronic integrator, as described in Section III of Annex VI to Commission Regulation
European Economic Community, no. 72/77.

6th procedure

06.01 Preparation of methyl esters of fatty acids from approximately 400 mg of the oil or fat component
test sample, prepare a solution containing about
20-50 mg / ml fatty acid methyl esters in n-hexane
method described in Section II, paragraph. Annex 3 VI to Commission Regulation European economic Community
no. 72/77.

6.2 TLC

6.2.1. Preparation of plates

A 500 mL round bottom flask was transferred onto 60 g silica gel (4.3)
added 120 ml of silver nitrate solution (4.5) and shaken for 1 min to
a fully homogeneous slurry. This suspension is then applied
conventional manner on plates. The thickness of the layer should be approximately 0.5 mm.
This quantity of slurry is sufficient for the preparation of five plates of
dimensions 200x200 mm.

Plates are allowed to partially air-dry (preferably in the dark for
about 30 min). The plates are completely dried and activated in an oven for 2.5 hours at 100
Wed. C. After activation, the board shall as soon as possible or
harbor in the dark before use and are re-activated.
Is sufficient activation at 110 st. C for 1 h, unless the plates are not darkened.
Before use in the deposited layer of sorbent Score lines through 10 mm from
side edges and the top of each plate, so that during development
reduce edge effects.

6.2.2. Application of methyl esters

Applicator (5.1.5.) Is applied to a narrow, about 50 mm long strip
least 40 mm from the edge and 10 mm from the lower edge 50
.mu.L solution of methyl esters (6.1) prepared from sample.
A similar manner to the plate 100 .mu.l of a mixed solution containing equal volumes
prepared solution of methyl esters (6.1) and a solution
erucate (04/06). Due to the fragility of the deposited layer of sorbent
proceed when depositing solutions particularly carefully. At the plate can also be applied
possibly 50 .mu.l of a solution
erucic acid (4.6) after development to assist in identifying the methyl strip
erucic acid. After application of the methyl with the bottom edge of the plate
in diethyl ether until the ether ascends to about 5 mm above the zone
sample application. So methyl esters are concentrated in a narrow strip.

6.2.3. Development of the plates

Into the development chamber Pour developing solvent to a height of about 5 mm
(4.8) and the chamber closed lid, in a refrigeration unit (5.1.1.)
Maintained at -25 st. C or as close to that temperature.
In some cases it may be appropriate to line the tank.
After two hours, the plate carefully in the tank and allow the solvent to
ascend to about one half to two thirds of the height plate.
Plate is removed and the solvent from it gently evaporated in nitrogen flow. the board is

Reinserted into the chamber and allow the solvent to ascend to the top
edge of the plate. Remove the plate and as previously dry in a nitrogen stream
then carefully sprayed with a solution
2,7-dichlorofluorescein (4.9).

View the plate under ultraviolet light, and the band containing
methyl erucate in the sample is determined by the highlight bar
sample to which was added methyl erucate.

6.2.4. Distribution of methyl

Methyl erucate band derived from the sample
scraped into a 50 ml beaker so as to avoid losses. Similarly to
another 50 ml beaker is transferred silica gel located above and below the strip
methyl erucate. This band will contain all the other factions
fatty acid methyl esters. To each beaker 1.0 ml of standard solution
methyl tetracosanoate (4.7) and 10 ml
diethyl ether (4/1). The contents of the beakers are mixed and transferred to
separate columns or funnels (5.1.4.), Each containing about 1 g
silica (4.4). The methyl esters were extracted with three or four 10 ml portions of diethyl ether
and the eluates are collected in small flasks. Each
filtrate was evaporated to a small volume under a stream of nitrogen and the methyl esters to small
tubes with conical bottom. Residual solvent was
evaporated in nitrogen flow so that the methyl esters concentrate at the bottom
tubes. Methyl esters are dissolved in 25-50 .mu.l of n-hexane (4/2).

06.03 Gas-liquid chromatography stationary phase

6.3.1. The procedure described in Section III of Annex VI to Regulation
Commission of the European Economic Community, no. 72/77 and tries to 1-2
.mu.L methyl ester solutions obtained from fractions containing methyl
erucic acid and the fractions containing the remainder of the methylated
fatty acids.

6.3.2. Electronic integrator at the following peak areas:

- From the chromatogram of the fraction containing methyl erucate surface
peaks methyl erucate (E), an internal standard (L1) of methyl
excluding internal standard (EF)

- From the chromatogram of the fractions containing the remainder of the methylated fatty acids
peak area of ​​internal standard methyl outside
(RF) and internal standard (L2).

7th Expression of results

07.01 Method of calculation and formula

7.1.1. Erucic acid content of the sample expressed as a percentage
methyl erucic acid of the total fatty acid methyl esters
prepared from the sample is given by:
E
-------------------- X 100
+ - - +
| EF RF |
| - + - |
L1 | L1 L2 |
+ - - +

Where E, EF, RF, L1 and L2 are the peak areas according to 6.3.2., If necessary, corrected
calibration factors.

Contents methyl erucate given by the above formula corresponds
erucic acid expressed as a percentage of total
fatty acids in the sample.

7.1.2. If peak areas are obtained in percentages can
values ​​for EF and RF may be calculated as follows: EF = 100
-
L1 RF = 100 - L2

7.1.3. The method of calculation 7.1.1. assumes that the quantity
tetracosanoic acid in the sample is negligible. If it turns out that
sample significant amounts of this acid, the acid value
tetracosanoic (L2) obtained from the chromatogram of the fractions containing the remainder
fatty acid methyl esters must be reduced as follows:

L2 - T2

where

T0P2
T2 = -----
P0

T2 is the peak area of ​​the methyl tetracosanoate
derived from the sample, forming part of the peak area
internal standard in the chromatogram of the remaining fraction
fatty acid methyl esters
P2 is the peak area of ​​methyl palmitate
chromatogram of the remaining fraction
T0 is the peak area of ​​the methyl tetracosanoate
from the chromatogram of the methyl esters of fatty acids
testing method established in accordance with Article 2 of the Directive
80/891 / EEC
P0 is the peak area of ​​methyl palmitate
from the chromatogram of the methyl esters of fatty acids
testing method established in accordance with Article 2 of Directive
Commission no. 80/891 / EEC.

7.1.4. Derivation formula

Proportion of fatty acids in the fraction containing methyl
erucic expressed as a percentage of the total fatty acids in
sample is given by:

EF -----

L1 EF

------------ ----------------- X 100 or 100 x
EF RF + - - + ----- + ---
| EF RF |
L1 L2 L1 + ----- + --- |
| L1 L2 |
+ - - +

Erucic acid in the fraction containing methyl
erucic is given by:

E
-----
EF

From erucic acid content of the sample expressed as a percentage of
total fatty acids is given by:

EF EE
X ----------------- ---- ----------------- x 100 x 100
+ - - EF + + - - +
| EF RF | or | EF RF |
L1 + ----- + --- | L1 + ----- + --- |
| L1 L2 | | L1 L2 |
+ - - + + - - +

7.1.5. repeatability

The difference between the results of two determinations carried out simultaneously
or in rapid succession on the same sample by the same operator under the same conditions
shall not be greater than 10% of the final value or 0.5 g per 100 g of sample
. The higher the value.


Příl.12
Testing methods to verify the composition of certain sugars intended for human consumption


First Sample preparation for the exam

Sample received by the laboratory thoroughly. For the test, the sample separates
amount of at least 200 g and transfer immediately to a clean, dry
, waterproof container fitted with airtight seal.

Second Reagents, instruments and aids

In describing the devices and accessories are referred for special
equipment and devices, or devices that must meet the specific requirements
.

If the mentioned water mean distilled water or demineralized water
comparable purity.

All reagents must be of analytical grade unless stated otherwise
.

Where reference is made to a reagent solution without further qualification, it is a
aqueous solution.

Third Expression of results

The test report shall state the result of the mean value obtained from two determinations
with satisfactory reproducibility.

Unless stated otherwise, the results are expressed in percentage weight of the initial
laboratory sample as it was in the laboratory
delivered.

Number of significant figures in the result so expressed by the precision
methods.


Příl.13
Method for determining the loss on drying to certain sugars intended for human consumption


First Subject matter and use

The method used to determine the loss on drying

- In semi-white sugar,

- In sugar or white sugar,

- The extra white sugar.

Second definition

Loss on drying means the value of the loss on drying
determined by the method specified.

Third Principle

Loss on drying is determined by drying at (103 +/- 2) st.
C.

Fourth Apparatus

04.01 Analytical balance accurate to 0.1 mg.

04.02 Drying with proper ventilation, thermostatically controlled, allowing
maintain temperature (103 +/- 2) st. C.

04.03 The metal pan with a flat bottom, resistant to the sample and
test conditions, with a diameter of 100 mm and a depth of at least 30
mm.

04.04 Desiccator with freshly activated silica gel or equivalent desiccant
with a moisture content indicator.

Fifth procedure

Note:

Operations described in section 5.1 to 5.7 must be performed immediately after
opening the sample container.

05.01 Dish (4.3) is dried to constant weight in an oven (4.2)
at (103 +/- 2) st. C.

Dish is left in a desiccator (4.4) to cool for at least 30 to 35
minutes and then weighed to the nearest 0.1 mg. To the bowl
weigh approximately 20-30 g sample with an accuracy of 0.1 mg.

The dish is placed in an oven (4.2) at (103 +/- 2 st. C), where
kept for 3 h.

Dish to cool in a desiccator (4.4) and weigh to the nearest 0.1 mg
.

Dish reinserted for 30 minutes in oven at (103 +/- 2) st.
C.

Allow to cool in the desiccator (4.4) and weigh to the nearest 0.1 mg
. If the difference between the two weighings greater than 1 mg, the procedure is repeated
. Where as weight, used to calculate
lowest recorded value.

The total drying time should be no longer than four hours.

6th Expression of results

06.01 Formula and method of calculation


Loss on drying weight percentage of the sample is given by the formula
:
(M0 - M1)
----------- X 100
m0
wherein:
m0 is the initial mass of the test sample (g)
m1 is the mass of the test sample after drying (g).

06.02 repeatability

The difference between the results of two determinations carried out simultaneously or in rapid succession on the same
same sample under the same conditions must be
greater than 0.02 grams per 100 grams of sample.


Příl.14
Method for determining the solids to certain sugars intended for human consumption

I.

Method of drying in a vacuum oven

First Subject matter and use

The method used to determine the solids content

- In starch syrup

- Dried glucose syrup,

- In monohydrate glucose

- In anhydrous glucose.

Second definition

Solids content means the solids content determined by the method.

Third Principle

Dry matter is determined at (70 +/- 1) st. C in a vacuum oven at
pressure of not more than 3.3 kPa (34 mbar). Test specimens of starch syrup or glucose syrup
dried before drying is adjusted by mixing with water and
kieselguhr.

Fourth Reagents

04.01 Diatomaceous earth: purified buchner
repeated washing with dilute hydrochloric acid (1 ml of concentrated acid
density of 1.19 g / ml per liter water at 20 st. C) until the filtrate showed
clearly acid reaction . Kieselguhr filter is then washed with water
until pH of filtrate rises above 4; then kieselguhr
dried in an oven at (103 +/- 2) st. C and stored in an airtight container
.

Fifth Apparatus

05.01 Vacuum drying oven, sealed, thermostatically controlled, equipped with a thermometer and a vacuum manometer
. It must be constructed so as to ensure
rapid heat transfer into the bottles stored on shelves.

05.02 Apparatus for drying air forms a glass column packed
freshly activated silica gel or equivalent desiccant, with
content indicator. This column containing concentrated sulfuric acid
is serially connected to the scrubber
connected to the inlet air to the dryer.

05.03 A vacuum pump capable of being controlled in an oven pressure of 3.3 kPa (34 mbar) or lower
.

05.04 The metal pan with a flat bottom resistant to
samples and test conditions with a diameter of 100 mm and a depth of at least 300 mm
.

05.05 Glass rod of a length such that it can not fully fit into
weighing bottle.

06.05 Desiccator with freshly activated silica gel or an equivalent desiccant
with a moisture content indicator.

07.05 Analytical balance accurate to 0.1 mg.

6th procedure

06.01 In a weighing bottle (4.5) with a glass rod (5.5) is converted
approximately 30 g of silica (1/4), all placed in an oven (1.5) with
temperature (70 +/- 1) st. C and the pressure reduced to less than 3.3 kPa (34 mbar).

Is dried for at least 5 h, while through an apparatus for drying air into the dryer
introducing a slow stream of air. Sometimes
pressure checked and adjusted as necessary.

06.02 The atmospheric pressure in the oven is reached again cautiously increase
dry air supply. Dish and the glass rod immediately
transferred to a desiccator (5.6) where it is allowed to cool, and then weighed
.

06.03 Beaker 100 ml with an accuracy of 1 mg
weigh approximately 10 g of the test sample.

06.04 The test sample is diluted with 10 ml of warm water and the solution using
glass rod (5.5) quantitatively transferred to a weighing bottle.

06.05 The dish with the test sample and the glass rod is inserted into
dryer and the pressure reduced to less than 3.3 kPa (34 mbar). Dried at (70 +/- 1
) st. C while being passed through the oven a slow stream of dry air
.

Drying was carried out for 20 hours; The greatest reduction in moisture
has come to the end of the first day. It is necessary to keep the vacuum pump running at
set pressure and allowed to slowly flow into the oven dry air so
that during the night the pressure maintained at approximately 3.3 kPa (34 mbar)
or lower.

06.06 Atmospheric pressure in the oven is reached again cautiously increase
dry air supply. The weighing bottle and its contents immediately transferred to
desiccator was allowed to cool, and then weighed to the nearest 1 mg
.

07.06 Operation (5.6) is repeated after a further 4 hours. The oven is restored

Atmospheric pressure and the dish was immediately placed in a desiccator. Allow to cool and weighing
determines whether there has been a constant
weight. Under constant weight is considered such an outcome when
difference between the two weighings of the same dish does not exceed 2 mg. Otherwise
case of repeated operations 07.06

08.06 To determine the dry matter in samples anhydrous glucose or glucose monohydrate is not
diatomaceous earth and water needed.

7th Expression of results

07.01 Formula and method of calculation The dry matter content expressed as a percentage
weight of the sample is calculated using the following formula:
100
(M1 - m2) x -----
m0
wherein:
m0 is the initial mass of the test sample (g), m1 is the mass
pan with kieselguhr, the glass rod and the rest
test sample after drying (g), m2 is the mass
weighing bottle with diatomaceous earth and glass rod (g).

07.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid
row of the same sample by the same, under the same conditions must be
more than 0.12 g per 100 g of sample.


Příl.15
Method for determining the total solids to certain sugars intended for human consumption
(refractometric method)

First Subject matter and use

Method provides a dry matter content

- Liquid sugar,

- Liquid white sugar

- Liquid invert sugar,

- White liquid invert sugar,

- In invert sugar syrup,

- In syrup from white invert sugar.

Second definition

Solids content means the solids content determined by the method.

Third Principle

An index of refraction of the test sample at 20 st. C, according to the tables in
listed in the Annex. 39 is converted to a dry matter content.

Fourth Apparatus

4.1 refractometer with a reading accuracy of four decimal places, equipped
thermometer and circulating water pump connected to a water bath, which is maintained
thermostatically controlled temperature (20 +/- 0.5) st. C.

4.2 Light source consisting of a sodium vapor lamp.

Fifth procedure

5.1 If present in the sample crystals and dissolved by diluting the sample
in a weight ratio of 1: 1st

5.2 refractometer (1.4) measure the refractive index of the sample at 20 Wed. C.

6th Expression of results and their calculation

Dry matter content is calculated from the refractive indices for sucrose solutions at 20 st.
C according to the table, and a correction for the presence of invert sugar
e test sample to the result of the tables added to each 1%
invert sugar value 0.022.

If the sample was diluted with water in a weight ratio of 1: 1, the content
calculated dry weight multiplied by two.

7th repeatability

Difference between the results of two determinations carried out simultaneously or in rapid
row of the same sample by the same, under the same conditions
not be greater than 0.2 g of dry matter per 100 g of sample.


Příl.16
Determination method of reducing sugars expressed as invert sugars
(method according to the research institute Institut Berlin)

First Subject matter and use

The method used to determine the reducing sugars expressed as invert sugar
semi-white sugar.

Second definition

Reducing sugar expressed as invert sugar content means
reducing sugars determined by the method.

Third Principle

Sample solution containing reducing sugars is used to reduce the cupric complex
solution. The resulting copper oxide is then oxidized with iodine solution
of known concentration, the excess of which is determined by back-titration
volumetric sodium thiosulfate solution of known concentration.

Fourth Reagents

04.01 Copper solution (Muller's solution)

4.1.1. In 400 ml of boiling water was dissolved 35 g of copper sulphate pentahydrate
(CuSO4 * 5H2O), and allowed to cool.

4.1.2. In 500 ml of boiling water was dissolved 173 g of potassium sodium tartrate tetrahydrate
(Rochelle salt KNaC4H4O6 * 4H2O) and 68 g
anhydrous sodium carbonate and allowed to cool.

4.1.3. Both solutions (4.1.1. And 4.1.2.) To a liter volumetric flask
and add water to 1 liter. After adding 2 g of activated carbon was
contents shaken, allowed to stand for several hours and then filtered through
thick paper or a membrane filter.

If during storage solution for a small amount of carbon
cuprous must filter again.

04.02 Acetic acid, a solution of 5 mol / l.


04.03 Iodine solution with a concentration of 0.01665 mol / l (4.2258 g / l).

04.04 Sodium thiosulfate at a concentration of 0.0333 mol / l.

05.04 Starch solution: per liter of boiling water is poured into a mixture of 5 g
soluble starch slurried in 30 ml of water, boiled for 3 min and allowed to cool
. If necessary, add 10 mg of mercuric iodide as
preservative.

Fifth Apparatus

05.01 Erlenmeyer flask, 300 ml; precision burettes and pipettes.

05.02 Water bath, boiling.

6th procedure

06.01 To a 300 ml Erlenmeyer flask is weighed portion of sample (10 g or less
) containing not more than 30 mg of invert sugar, and dissolved
in about 100 ml of water.

To the flask with the sample solution Pipette 10 ml of the copper solution
(1.4), the content was swirled to mix and the flask was placed in a boiling water bath
(5.2) for exactly 10 minutes. The level of the solution in the Erlenmeyer flask
must be at least 20 mm below the water level in a water bath
. The flask is rapidly cooled with cold water, the solution
not stir to prevent reoxidation of the precipitated cuprous oxide
air oxygen.

Without shaking the contents of pipette 5 ml of acetic acid solution
(2.4) 5 mol / l as soon burette was added an excess (20
40 mL) solution of iodine (3.4) at a concentration 0.01665 mol / l.

Precipitate copper dissolved by stirring and the excess iodine is titrated with sodium thiosulfate solution
(4.4) at a concentration of 0.0333 mol / l when using
starch solution (4.5) as indicator. The indicator is added to the end
titration.

06.02 With water, the blind test, which is repeated every time a new application
copper solution (4.4). Titration consumption does not exceed 0.1 ml.

06.03 The sugar solution in the cold, performs a control test.
The solution was allowed to stand at room temperature for 10 min to allow for
reacting other, possibly present reducing agents, such as sulfur dioxide
.

7th Expression of results

07.01 Formula and method of calculation

Volume of iodine solution equal to the volume (ml) of excess
added a solution of iodine (0.01665 mol / l) minus the volume (ml) solution
thiosulphate solution (0.0333 mol / l) consumed in the titration .

Volume (ml) of iodine solution (0.01665 mol / l) is adjusted by subtracting
:

7.1.1. the number of milliliters in the blank test with water (6.2)

7.1.2. the number of milliliters in the control test with a sugar solution
cold (6.3)

7.1.3. volume of 2.0 ml per each 10 g of sucrose present in the aliquot used
or proportional amount, if the sample contains less than
10 g sucrose (correction for sucrose).

After these corrections corresponds to the consumption of 1 ml of iodine solution
(3.4) 1 mg of invert sugar.

Invert sugar content percentage of the sample is calculated according to the formula:
V1
------
10 x m0

wherein:
V1 - the number of ml of iodine solution (3.4) after correction,
m0 - the weight of sample (g).

07.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid
row of the same sample by the same, under the same conditions
not be greater than 0.02 g per 100 g of sample.


Příl.17
Determination method of reducing sugars expressed as invert sugar
(Method according to Knight and Allen)

First Subject matter and use

The method used to determine the reducing sugars expressed as invert sugar


- In sugar or white sugar,

- The extra white sugar.

Second definition

Reducing sugar expressed as invert sugar content means
reducing sugars determined by the method.

Third Principle

The sample solution was added excess reagent and a reduced and
nezredukovaný portion was then determined by back titration with sodium copper
disodium ethylendiamintetraoctvé.

Fourth Reagents

04.01 Disodium salt of ethylenediaminetetraacetic acid, a solution of 0.0025
mol / l: Dissolve 0.930 g of disodium salt of ethylenediaminetetraacetic acid
in water and make up with water to one liter.

04.02 Murexide indicator solution: 0.25 g Murexide was added to 50 ml of water
and mixed with 20 ml of aqueous methylene blue solution
0,2 g / 100 ml.

04.03 Copper reagent: in 600 ml of water containing 40 ml of sodium hydroxide
of 1.0 mol / l were dissolved 25 g of anhydrous sodium carbonate and

25 g potassium sodium tartrate tetrahydrate. In about 100 ml of water was dissolved
6.0 g of copper sulphate pentahydrate, the resulting solution was added to a solution
tartrate and supplemented with water to one liter. Note: The solution has a limited shelf life
(one week).

4.4 Standard solution invert in a volumetric flask of 250 ml
Dissolve 23,750 g of pure sucrose (4.5) in about 120 ml of water.
Added 9 ml of hydrochloric acid (? 20 = 1.16) and allowed to stand at
room temperature for 8 days. The solution is made up to 250 ml and
termination of hydrolysis the check reading polarimeter or
saccharimeters using a tube with a length of 200 mm. The measured value is equal to a
(11.80 +/- 0.05) st. S (see note). 200 ml of this solution
pipetted into a volumetric flask of 2000 ml, diluted with water and
continuous shaking (to avoid excessive local basification
solution) were added 71.4 ml of sodium hydroxide solution (1 mol / l) in
which are dissolved 4 g of benzoic acid. The solution is made up to 2000
ml; ie. to contain 1 g of invert sugar in 100 ml of pH
solution is to move past the third

This stable stock solution was diluted just before use.

05.04 Pure sucrose: sample of pure sucrose with an invert sugar content
exceeding 0.001 g / 100 g.

Fifth Apparatus

5.1 Tubes 150 x 20 mm.

5.2 White porcelain bowl.

5.3 Analytical balance accurate to 0.1 mg.

6th procedure

06.01 In a tube (5.1) was dissolved 5 g of the sample sugar in 5 ml of cold
water, add 2.0 ml of copper reagent (4.3) and mix.
Tube was immersed in a boiling water bath for 5 minutes and then in cold water
cooled.

06.02 The solution from the tube quantitatively, using the smallest possible amount of water
converted into white porcelain dish (5.2), add three
drops of indicator (2.4) and titrated with a solution of disodium ethylenediaminetetraacetic
( 1.4). Titration consumption in ml is designated as
Vo.

Just before the end of the titration the color of the solution changes from green through gray to purple
at equivalence point. Purple color, slowly disappearing in
due to oxidation of copper oxide to copper oxide oxidation rate
depends on the concentration of reduced copper present. Therefore, the titration must
reach the equivalence point as possible.

06.03 Plot a calibration curve based on the addition of a known quantity
invert sugar (appropriately diluted solution 4.4) to 5 g of pure sucrose
(4.5) and the corresponding amount of cold water to bring the total volume
solution added was 5 ml. Titration consumption (ml) plotted against percentage
; invert sugar content added to 5 g of sucrose
resulting curve is a straight line in the range of 0.001 to 0.019 g per 100 g
invert sugar, respectively. 100 g of sample.

7th Expression of results

07.01 The calculation procedure

From the calibration curve the invert sugar content (in percent)
corresponding consumption in ml of disodium salt of ethylenediaminetetraacetic acid
when testing sample.

07.02 If it is assumed a higher concentration than 0.017 g invert sugar
in 100 g of the test sample must be reduced correspondingly
quantity of sample in section 6.1., The test specimen, however, must be added to 5 g of pure
sucrose (4.5 ).

07.03 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid
row of the same sample by the same, under the same conditions
not be greater than 0.005 g per 100 g of sample.

8th Note

When converting values ​​in st. With at polarimeter with angular degrees
value specified in st. The divide 2,889 (polarimeter tubes with a length of 200 mm;
sodium lamp as the light source, the device placed in the room
which it is possible to keep the temperature around 20 degrees. C).


Příl.18
Determination method of reducing sugars expressed as invert sugar, or glucose
equivalent (Method Luff-Schoorl)

First Scope and field of application method is used to determine

01.01

Reducing sugar content expressed as invert sugar

- Liquid sugar,

- Liquid white sugar

- Liquid invert sugar,

- White liquid invert sugar,

- In invert sugar syrup,

- In invert sugar syrup white.

02.01

Reducing sugar content expressed and calculated (based on dry weight
) as the glucose equivalent


- In starch syrup

- Dried glucose syrup.

03.01

Reducing sugar content expressed as D-glucose

- In glucose monohydrate

- In the anhydrous glucose.

Second definition

Reducing sugars expressed as invert sugars, D-glucose or glucose
equivalent means reducing sugar content expressed or
calculated as invert sugar, D-glucose or glucose equivalent
determined by the method.

Third Principle

Sample with reducing sugars are heated (and if necessary clear)
under standard conditions to the boiling point with a copper solution, which
partially reduced to Cu (I). Excess Cu (II) is then determined iodometrically
.

Fourth Reagents

04.01 Carrez solution I: 21.95 g of zinc acetate dihydrate, or trihydrate 24 g
zinc acetate, is added together with 3 ml of glacial acetic acid
dissolved in water and made up with water to 100 ml.

04.02 Carrez II solution: 10.6 g ferrocyanide trihydrate
potassium is dissolved in water and made up with water to 100 ml.

04.03 Luff-Schoorl reagent: prepare the following solutions:

4.3.1. Copper sulphate solution: 25 g of copper sulfate pentahydrate
free from iron was dissolved in 100 ml of water.

4.3.2. Citric acid solution: 50 g of citric acid monohydrate
was dissolved in 50 ml of water.

4.3.3. Sodium carbonate solution: 143.8 g of anhydrous sodium carbonate
dissolved in about 300 ml of hot water and allowed to cool.

4.3.4. To a sodium carbonate solution (4.3.3.) In a one liter volumetric flask
mild stirring swirl added a solution of citric acid
(4.3.2.). The contents were stirred until no effervescence
then a solution of copper sulfate (4.3.1.) And water to make up to 1000 ml
. The solution was allowed to stand overnight, if necessary, then
filtered. A check is performed according to the concentration of the reagent solution
method described in section 6.1. (Cu; 0.1 mol / l sodium carbonate 1 mol / l).

04.04 Solution of sodium thiosulfate, 0.1 mol / l.

05.04 Starch solution: to one liter of boiling water is poured into 5 g
soluble starch slurried in 30 ml water. Boil for 3 minutes and allowed to cool
; if necessary, added with 10 mg of mercury iodide
as preservative.

06.04 Sulfuric acid, 3 mol / l.

07.04 A solution of potassium iodide, 30% by weight.

08.04 Fragments of pumice stone, boiled in hydrochloric acid, washed with water until disappearance
acidic and dried.

09.04 Isopentyl.

4.10 Sodium hydroxide, 0.1 mole / l.

4.11 Hydrochloric acid, 0.1 mol / l.

4.12 Phenolphthalein, 1% solution in ethanol.

Fifth Apparatus

05.01 Erlenmeyer flask 300 ml with a reflux condenser.

05.02 Stopwatch.

6th procedure

06.01 Titer determination Luff-Schoorl reagent (4.3)

6.1.1. To 25 ml Luff-Schoorl reagent (3.4) were added 3 g of potassium iodide
and 25 ml of sulfuric acid at a concentration of 3 mol / l (4.6).

Titrate with sodium thiosulphate 0,1 mol / l (4.4)
using the starch solution (5.4) as indicator, added
until the end of the titration. If there is no consumption of the sodium thiosulphate concentration
0.1 mol / l equal to 25 ml, it must be re-prepared agent.

6.1.2. Pipette 10 ml of reagent into a 100 ml volumetric flask and dilute to the mark with water
.

Thus 10 ml of the diluted reagent was pipetted into an Erlenmeyer flask containing 25 ml
hydrochloric acid 0.1 mol / l
(4.11) and heated for one hour in a boiling water bath. Then
solution was cooled, made up to the original volume with freshly boiled water and
titrated with a solution of sodium hydroxide of 0.1 mol / l (4.10.) To
using phenolphthalein (4.12) as indicator.

Consumption of sodium hydroxide solution, 0.1 mol / l (4.10) must be between 5.5 and 6.5 ml
.

6.1.3. 10 ml of diluted reagent (6.1.2.) Is titrated acid
HCl, 0.1 mol / l (4.11.) Using phenolphthalein (4.12)
as indicator. The equivalence point during titration is characterized by loss
violet color.

Consumption of hydrochloric acid, 0.1 mol / l (4.11) must be
between 6.0 and 7.5 ml.

6.1.4. The pH of the Luff-Schoorl reagent must be between 9.3 and 9.4 at 20
Wed. C.

06.02 Preparation of the solution

6.2.1. Weigh 5 g sample with an accuracy of 1 mg and
quantitatively transferred to a volumetric flask of 250 ml containing 200 ml of water. IN

If necessary, clarified by adding 5 ml of Carrez solution I (4.1), then
was added 5 ml of Carrez solution II (2.4). After the addition of each solution
content is mixed. The solution is made up with water to 250 ml and mix well. In
If necessary, the solution is filtered.

6.2.2. Solution (6.2.1.) Were diluted in a ratio such that the content
reducing sugars expressed as glucose in 25 ml
ranged from 15 to 60 mg.

06.03 Titration by the Luff-Schoorl method

To a 300 ml conical flask (1.5) Pipette 25 ml
Luff-Schoorl reagent (4.3), into the flask were then Pipette 25 ml of
sugar solution (6.2.2.) And inserted two fragments of pumice stone (4.8).
The flask (1.5) with a reflux condenser and equipment were immediately placed on
asbestos wire gauze over a Bunsen burner flame. The mesh has
asbestos parts cut a circular hole of the same diameter as the bottom
flask. The fluid is approximately two minutes to the boil and allowed to
boil gently for 10 minutes. Cool immediately in cold water and after 5
min is titrated according to the following procedure:

Add 10 ml potassium iodide solution (4.7), immediately followed by
added cautiously (considering effervescing), 25 ml of sulfuric acid
concentration of 3 mol / l (4.6). Titrate with sodium thiosulphate solution
about 0,1 mol / l (4.4) until the solution is almost decolourised then added as an indicator
few ml of starch solution (4.5) and continue
titration until the blue color disappears. Carry out a blank test with 25 ml
water instead of 25 ml of sugar solution (6.2.2.).

7th Expression of results

07.01 Formula and method of calculation

The table below is deducted or down interpolation weight
glucose or invert sugar in mg corresponding to the difference between the two titration
fuel consumption, expressed in ml of sodium thiosulphate
about 0.1 mol / l.

The result is expressed in weight percent invert sugar or D-glucose
, based on dry matter.

07.02 repeatability

The difference between the results of two determinations carried out simultaneously or in rapid
themselves from the same sample by the same operator under the same conditions
not be greater than 0.2 ml.

8th Note

Before acidification with sulfuric acid may be added a small amount
isopentyl alcohol (4.9) to reduce foaming.
Table of values ​​for Luff-Schoorl reagent

----------------------------------------------- -----------
0.1 mol / l sodium thiosulfate
glucose, fructose, invert sugar, C6H12O6

------------ ----------------------------------------------
ml Unlike mg
---------------------------------------------
------------- 1 2.4
2 4.8 2.4
3 7.2 2.4
4 9.7 2.5
5 12.2 2.5
6 14.7 2.5
7 17.2 2.5
8 19.8 2.6
9 22.4 2.6 10 25.0 2.6


11 27.6 2.6 12 30.3 2.7 13 33.0 2.7

14 35.7 2.7 15 38.5 2.8


16 41.3 2.8 17 44.2 2.9 18 47.1 2.9

19 50.0 2.9 20 53.0 3.0


21 56.0 3.0 22 59.1 3.1 23 62.2 3.1

-------------------------------------------------- --------


Příl.19
Determination method of reducing sugars expressed as invert sugar
(Method of the Lane and Eynon method - modification of constant volume)

First Subject matter and use

The method used to determine the reducing sugars expressed as invert sugar


- Liquid sugar,

- Liquid sugar, white,

- Liquid invert sugar,

- White liquid invert sugar,

- In invert sugar syrup,

- In invert sugar syrup white.

Second definition

Reducing sugar expressed as invert sugar content means
reducing sugars determined by the method.

Third Principle

Test sample solution is titrated at the boiling point of a certain quantity
Fehling's solution, using methylene blue as indicator.

Fourth Reagents

04.01 Fehling's solution:

4.1.1. Solution A: 69.3 g of copper sulphate pentahydrate is dissolved in
water and make up to 1000 ml.

4.1.2. Solution B: 346.0 g sodium potassium tartrate tetrahydrate and 100 g
sodium hydroxide were dissolved in water and make up to 1000 ml.
Clear solution was decanted from the precipitate, which can sometimes create.


Note: These two solutions should be stored in brown or amber
amber-colored bottles.

04.02 Sodium hydroxide solution, 1 mol / l.

04.03 Standard solution of invert sugar: 23,750 g of pure sucrose
was dissolved in about 120 ml of water in a 250 ml volumetric flask, add 9 ml
hydrochloric acid (density 1.16 g / ml) and allowed to stand at || | room temperature for 8 days. The solution is made up to 250 ml and the hydrolysis was complete
checked polarimeter or saccharimeters with
tube length of 200 mm. The reading should be equal to (11.80 +/- 0.05)
st. S (see Note 8). 200 ml of this solution was pipetted into a 2000 ml volumetric flask
, diluted with water, and while shaking (to avoid excessive local
alkalizing solution) were added 71.4 ml of sodium hydroxide solution
1 mol / l (2.4) in which are dissolved 4 g
benzoic acid. Make up to 2000 ml to obtain a solution containing 1 g
invert sugar per 100 ml. The pH of the solution should be approximately equal
3. This stable stock solution should be diluted to
before use. In the preparation of invert sugar solution by
concentration of 0.25 g / 100 ml 250 ml volumetric flask is filled to volume
stock solution having a concentration of 1 g / 100 ml at 20 Wed. C. The content
quantitatively transferred into a 1000-mL volumetric flask and dilute with water to
mark at 20 st. C.

04.04 Methylene blue solution, 1 g / 100 ml.

Fifth Apparatus

05.01 Bottom flask with a narrow neck 500 ml.

05.02 Burette 50 ml, graduated in 0.05 ml with the lateral valve.

05.03 Pipettes with markings 20, 25 and 50 ml.

05.04 Volumetric flask of 250, 1,000 and 2,000 ml.

05.05 Heating device, suitable for maintaining boiling under conditions specified
in section 6.1. and to monitor the color change at equivalence point,
without boiling flask (1.5) must be moved.

06.05 Stopwatch showing an accuracy of at least 1 s.

6th procedure

06.01 Titer determination Fehling's solution

6.1.1. To a clean, dry beaker Pipette 50 ml solution B (4.1.2.)
Then 50 ml of solution A (4.1.1.), And mix well.

6.1.2. Burette was flushed and filled with 0.25% (0.25 g / 100 mL)
standard invert sugar solution (4.3).

6.1.3. To a 500 ml round bottom flask (1.5) Pipette 20 ml aliquot
mixed solution A and B (6.1.1.) And poured with 15 ml of water. From the burette into the flask metered
39 ml invert sugar solution, add a few anti
stones and contents gently swirl mix.

6.1.4. The flask contents are heated to boiling and allowed to boil for exactly 2 min
. During the next steps, the flask must not remove from the heat and
interrupt var.

At the end of the two-minute boiling add three or four drops of solution
methylene blue (4.4). The color of the solution must be bold blue.

6.1.5. Continue the addition of standard solution of invert sugar
burette, initially 0.2 ml then 0.1 ml and then added dropwise until
reach the equivalence point. This is indicated by the disappearance of blue coloration
present methylene blue. The color solution is reddish due
presence of a slurry of cuprous oxide.

6.1.6. Equivalence point during titration should be achieved within three minutes
from the start point of the solution. Final consumption V0 must move
ranging from 39.0 to 41.0 ml. If the value V0 outside the boundaries of this range,
adjust the concentration of copper in the Fehling's solution A (4.1.1.) And the determination is repeated
titer.

06.02 Preparation of sample

The concentration of the test sample solution should be in the range 250 to
400 mg of invert sugar in 100 ml.

06.03 Screening test

6.3.1. It is necessary to perform a screening test to determine the required
addition of water to 20 ml of the mixed solution A and B, which ensures that the final volume after titration
will amount to 75 ml. The procedure is identical with the instruction at point
6.1.4., But instead of the standard invert sugar solution is used
sample solution, i.e. to a flask with 25 ml burette metered sample solution
was added 15 ml of water, allow to boil for 2 min and the solution was then titrated to
to reach the equivalence point, as described in section 6.1.5.

6.3.2. If, after addition of methylene blue solution persists
reddish color is employed sample solution is too concentrated.
In this case the test is interrupted and carried repeat determination with
lower concentration of the sample solution. If to achieve
reddish coloration takes more than 50 mL of the sample solution must be used

Sample with a higher concentration.

Amount of water to be added is calculated by subtracting the volume
mixed Fehling's solution (20 ml) and the volume of the sample solution from 75 ml.

06.04 Final testing of the sample solution

6.4.1. Into the boiling flask Pipette 20 ml of mixed Fehling's solution and the amount of water set
6.3.

6.4.2. From the burette is metered sample solution in an amount determined by
point 6.3, reduced by 1 ml. Add a few glass beads,
contents of the flask was swirled to mix, boil and titrate as in the previous
stage (6.3). Equivalence point during titration should be reached within 1 min
by adding methylene blue solution. Consumption in the final titration
equals V1.

7th Expression of results

07.01 Formula and method of calculation The content of reducing sugars in the sample
expressed as invert sugar is calculated by the formula:
% reducing sugars (as invert sugar)

V0 x 25 xf ------------

C
x V1 Where
:

C - the concentration of the test sample solution (g / 100 mL)
V0 - consumption standard invert sugar solution (mL)
titer determination,
V1 - consumption of the test sample solution (ml) at a precise test
(6.4.2.)
F - correction factor reflecting the concentration of sucrose present
the test sample solution; values ​​are given below
table:
----------------------------------


Sucrose (g mixture) Correction ---------------------------------- factor f

0.0 1,000
0.5 0,982
1.0 0,971
1.5 0.962
2.0 0.954
2.5 0.946
3.0 0.939
3.5 0,932
4.0 0.926
4.5 0.920
5.0 0.915
5.5 0,910
6.0 0.904
6.5 0.898
7.0 0.893
7.5 0.888
8.0 0.883
8.5 0.878
9.0 0.874
9.5 0.869 10.0 0.864

---------------------------------- || |
Correction for different amounts of sucrose solution in a test sample
can be calculated from the tabulated values ​​by interpolation.

Note: The approximate sucrose concentration may be determined by subtracting
concentrations of dissolved solids from invert sugar (
for this calculation is the value used is 1.0 f) the concentration of total dissolved solids
expressed as sucrose, and
determined on the basis of the refractive index in accordance with Annex no. 15 hereto.

07.02 repeatability

The difference between the results of two determinations carried out simultaneously or in rapid
themselves from the same sample by the same operator under the same conditions
must not be greater than 1.0% of their arithmetic mean.

8th Note

When converting values ​​in st. With at polarimeter with angular degrees this
value reported in st. S divides factor 2.889 (polarimetric tube
length of 200 mm; sodium lamp as the light source, the unit placed
room in which it is possible to maintain a temperature close to 20 °. C).


Příl.20
Method for determining glucose equivalent to certain sugars intended for human consumption
(Method of the Lane and Eynon method with constant titre)

First Subject matter and use

This method is used to determine glucose equivalent

- In starch syrup

- Dried glucose syrup,

- In monohydrate glucose

- In anhydrous glucose.

Second definition

02.01 Reducing power means reducing sugars
determined by the method specified, expressed as anhydrous D-glucose and is measured in percentage
sample weight.

02.02 Glukosovým equivalent means reducing power, calculated in
percent of the dry weight of the sample.

Third Principle

Test sample solution under precisely defined conditions titrated
boiling certain volume of mixed Fehling's solution, using methylene blue
indicator.

Fourth Reagents

04.01 Fehling's solution:

4.1.1. Solution A:

The Dissolve 69.3 grams of copper sulfate pentahydrate and
volumetric flask 1000 ml Fill to the mark.

4.1.2. Solution B:

In water were dissolved 346.0 g tetrahydrate potassium sodium tartrate and 100
g of sodium hydroxide and in a volumetric flask of 1000 ml volume with water after
mark. The clear solution was decanted from the sediment, which sometimes can create
.

Note: These two solutions (4.1.1. And 4.1.2.) Should be stored in brown or amber
amber-colored bottles.


4.1.3. Preparation of the mixed Fehling's solution into a clean beaker
Pipette 50 ml solution B (4.1.2.), And then 50 ml of solution A (4.1.1.) And
well mixed.

Note: Mixed Fehling's solution should not be stored but must
prepared fresh every day and standardized (6.1).

04.02 Anhydrous D-glucose (C6H12O6) This material must before use
dry for 4 hours in a vacuum oven at 100 +/- 1 st. C or lower and
at about 10 kPa (103 mbar).

04.03 The standard solution of glucose, 0.600 g / 100 ml

With accuracy of 0.1 mg Weigh 0.6 g anhydrous glucose (2.4)
dissolve in water, transfer quantitatively into a volumetric flask of 100 ml
(4.5), dilute with water and mix.

This solution must be prepared fresh each day when it will be used.

04.04 Methylene blue solution, 0.1 g / 100 ml water

Dissolve 0.1 g methylene blue in 100 mL water.

Fifth Apparatus

05.01 Bottom flask with a narrow neck 250 ml.

05.02 Burette 50 ml, with a side stopcock and divided by 0.05 ml.

05.03 Pipettes not cut with notches 25 ml and 50 ml.

05.04 Volumetric flask of 100 and 500 ml.

05.05 Heating device, suitable for maintaining boiling in accordance with
conditions described in section 6.1. and to monitor color change
equivalence point without boiling flask (1.5) is removed from the heat source
(see section 6.1. Note 1).

06.05 Stopwatch showing an accuracy of at least 1 s.

6th procedure

06.01 Standardization of the Fehling's solution

6.1.1. To clean, dry boiling flask (1.5) Pipette 25 ml of Fehling
solution (4.1.3.).

6.1.2. Burette (2.5) is filled with a standard glucose solution (4.3) and
meniscus is set to the zero mark.

6.1.3. To a round bottom flask (1.5) from the burette impregnated with 18 ml of standard
glucose solution (3.4) and swirl the contents to mix.

6.1.4. Round bottom flask is placed on a heating device (5.5) previously
adjusted so that boiling occurred at (120 +/- 15). Throughout
titration may be heating device adjusted differently (note 1)

6.1.5. Once the solution begins to boil, start the stopwatch.

6.1.6. The flask contents are boiled for 120 seconds (by stopwatch). Toward the end
1 ml of methylene blue solution (4.4).

6.1.7. After 120 s, measured by a stopwatch, from the beginning of boiling, begins from the burette
(6.1.2.) To the boiling flask (1.5) adding a standard solution of glucose, 0.5 ml doses
until decolorized methylene blue ( see notes 2 and 3
).

Record the total volume of added standard solution of glucose (X
ml) including the penultimate 0.5 ml dose.

6.1.8. Instance in paragraphs 6.1.1. and 6.1.2. They are repeated.

6.1.9. From the burette to the boiling flask (5.1) fills (X - 0.3)
ml of standard glucose solution.

6 January 10. Instance in paragraphs 6.1.4., 6.1.5. and 6.1.6. They are repeated.

6 January 11. After 120 s, measured by a stopwatch, from the beginning of boiling, begins from the burette into
boiling flask (1.5) adding a standard solution of glucose, initially 0.2 ml batches
and finally dropwise until the methylene blue being | || colorless. At the end of this process must be the time between two consecutive
subsequent addition of standard solution of glucose 10 to 15 seconds.

This addition must be done within 60 hours, the total boil
therefore must not exceed 180 seconds.

For this purpose, it may be necessary to carry out the third titration with a slightly larger
, the initial dose adjusted accordingly
standard solution of glucose (6.1.9.).

6 January 12. Record the total consumption (V0 ml) of standard solution at
final titration (see note 4).

6 January 13. Value V0 should be between 19.0 to 21.0 ml
standard glucose solution (3.4).

If the value V0 is outside this range, appropriately adjusted
concentration of Fehling's solution A (4.1.1.) And the standardization process is
repeated.

6 January 14. Given that the value of V0 is precisely known, it is necessary to standardize
daily take one titration with initial addition of the standard solution of glucose
V0 - 0.5.

Note 1: This ensures that once boiling occurs, the steam generation
rapid and uninterrupted throughout the titration procedure, and in the most
to prevent entry of air into the titration flask and re-oxidation of its contents
.

Note 2: The discoloration of methylene blue solution is preferably
observable in the upper layers of the contents of the titration flask and in
meniscus, i.e. in areas, which are substantially free of clotted red

Copper oxide. Discoloration is more observable
for indirect lighting. It is preferable to use a diffuser for the titration flask.

Note 3: The burette should be avoided during the determination of the best protected from
heat source.

Note 4: Because they can not completely exclude the subjective factor
, every worker must carry out his own standardization titration
and the calculation has to use his own value of V0 (7.1).

06.02 Screening test sample prepared

6.2.1. If not known approximate value of reducing power (1.2)
prepared sample, it is necessary to perform a preliminary test to
acquired this approximate value, and could thus be calculated
weight of the test sample (3.6). This test is conducted as follows:

6.2.2. Prepared from% (m / v) solution of the sample, "Z" is an approximate value.

6.2.3. The procedure is as in 6.1.2. using a sample solution
(6.2.2.) instead of a standard glucose solution.

6.2.4. The procedure is as in 6.1.1.

6.2.5. The procedure is as in 6.1.3. with 10 ml of the sample solution
instead of 18.0 ml of the standard solution of glucose.

6.2.6. The procedure is as in paragraph 6.1.4.

6.2.7. The flask contents are heated to boiling. Add 1 ml of methylene blue
(4.4).

6.2.8. When it starts boiling, starts the timer (6.5) and will be added
sample solution from the burette into the flask 1.0 ml doses, at about 10
seconds, until the decoloration of methylene blue.

Recorded total consumption of standard solution of glucose (Y ml)
including last 0.5 ml dose.

6.2.9. The Y value should not exceed 50 ml. If this happens, you need
increase the concentration of the sample solution and repeat the titration.

6 February 10. Approximate value of reducing power of the prepared sample in% is given by approximately
:
60 x V0
---------
Y x Z

6.3 Test sample 0,1 mg dare so much
prepared sample (mg) which contains 2.85 to 3.15 g
reducing sugars expressed as anhydrous dextrose (D-glucose) . When calculating the
used either known approximate value of reducing power (1.2) or
approximate value obtained in point 6 February 10.

6.4 test solution the test sample in a volumetric flask of 500 ml
dissolved in water and make up to volume with water.
6.5 Determination


6.5.1 Operating as in 6.1.1.

6.5.2 burette (2.5) is filled with the test solution (4.6) and the meniscus
set to zero.

6.5.3 from a burette to the boiling flask fills 18.5 ml solution of the test
solution. Swirl the flask contents mixed.

6.5.4 proceeds as described in paragraph 6.1.4.

6.5.5 proceeds as described in paragraph 6.1.5.

6.5.6 proceeds as described in paragraph 6.1.6.

6.5.7 proceeds as described in point 6.1.7., Using test solution
instead of a standard glucose solution.

6.5.8 proceeds as described in paragraph 6.1.8.

6.5.9 proceeds as described in paragraph 6.1.9., Using test solution
instead of a standard glucose solution.

06/05/10 proceeds as described in paragraph 6 January 10.

05/06/11 proceeds as described in paragraph 6 January 11., Using test solution
instead of a standard glucose solution.

05/06/12 Note the volume (V1) of test solution consumed during titration
final.

05.06.13 The V1 be in the range of 19.0 to 21.0 ml of test solution
.

If the value of V1 is outside this range, adjust the concentration of the test solution and
grade 6.5.1. to 05/06/12. They are repeated.

06.05.14 Carry out two determinations using the same test solution.

6.6 Solids content Method 2 down the dry matter content of the prepared sample.

7th Expression of results

07.01 Formula and calculation procedure

7.1.1. Reducing power

Reducing power expressed in weight percent of the prepared sample is given by
:
300 x V0
----------
V1 x M

where
V0 - volume (ml) of standard glucose solution (3.4)
consumed in the standardization titration (6.1)
V1 - volume (ml) of the test solution (4.6) consumed during
titration in the assay procedure (5.6)
M - mass of the test sample (g) (6.3) used
to prepare 500 ml of the test solution.

7.1.2. Glucose equivalent

Glucose equivalent calculated the percentage by weight of dry matter
prepared sample is given by:
RP x 100
---------
D


Where RP - reducing power, calculated in percentages by weight
prepared sample (7.1.1.)

D - dry matter content of the prepared sample in percent by weight.

7.1.3. The result is the arithmetic mean of the two determinations
provided that it is satisfied with the requirement for repeatability (2.7).

07.02 repeatability

The difference between the results of two determinations carried out simultaneously or in rapid
themselves from the same sample by the same operator under the same conditions
must not be greater than 1.0% of their arithmetic mean.


Příl.21
The method of determination of sulphate ash for certain sugars intended for human consumption


First Subject matter and use

The method used to determine the content of sulfate ash

- In starch syrup

- Dried glucose syrup,

- In monohydrate glucose

- In anhydrous glucose.

Second definition

Sulfate ash content means the content of sulfate ash as determined
this method.

Third Principle

After combustion test sample in an oxidizing atmosphere at 525 st.
C in the presence of sulfuric acid is determined by its residual mass is
expressed in percentage weight of the sample.

Fourth Reagents

04.01 Sulphuric acid, dilute solution: slowly and carefully added 100
ml concentrated sulfuric acid (density at 20 °. C = 1.84 g / ml; 96%
wt.) In 300 ml of water while stirring and the cooling.

Fifth Apparatus

05.01 Electric muffle furnace, equipped with a pyrometer and capable of operating
at (525 +/- 25) st. C.

05.02 Analytical scales accurate to 0.1 mg.

05.03 Crucible for determination of ash, platinum or quartz, suitable
volume.

05.04 Desiccator containing freshly activated silica gel or equivalent desiccant with
content indicator.

6th procedure

Crucible (3.5) is heated to the combustion temperature, kept in a desiccator
cool and weigh. Do exactly the crucible (0.1 mg) Weigh 5 g
glucose syrup or dried glucose syrup, or 10 g
monohydrate glucose and anhydrous glucose. Add 5 ml solution
sulfuric acid (4.1) (see note 8.1) and the sample in the crucible was heated cautiously
flame or hotplate until completely
nezuhelnatí. Charring during which burns the steam from the sample (see note
8.2) must be carried out in a fume cupboard.

Crucible (3.5) was placed in a muffle furnace (1.5), preheated to (525 +/- 25
) st. C, and leave it until a white ash.
It normally takes two hours (see note 8.3).

Sample was about 30 minutes. cool in the desiccator (4.5) and then
weighed.

7th Expression of results

07.01 Formula and method of calculation

Sulfate ash content, in percentage by mass
sample to be tested is given by:
m1
S = --- x 100
m0


Where m1 - mass of ash (g)
m0 - mass of test sample (g).

07.02 repeatability

The difference between the results of two determinations carried out simultaneously or in rapid succession on the same
same sample under the same conditions must be
greater than 2.0% of their arithmetic mean.

8th Notes

08.01 Sulfuric acid is added in small portions to avoid excessive foaming
.

08.02 When the first combustion is necessary to proceed extremely carefully to avoid losses
sample or ash content due to excessive swelling of the sample.

08.03 If it is difficult to achieve complete combustion (i.e.
remain black particles) is necessary crucible from the muffle furnace to remove the residue after cooling
moistened with a few drops of water and re-inserted into the furnace.


Příl.22
Method for determining polarization to certain sugars intended for human consumption

First Subject matter and use

The method used to determine the polarization

- In semi-white sugar,

- In sugar or white sugar,

- The extra white sugar.

Second definition

Polarization means the ability of the sugar solution containing 26 g
sugar per 100 ml, which is closed in a tube of 200 mm in length, bottle
plane of polarized light.

Third Principle

Polarization is determined saccharimeter or a polarimeter
under the conditions described in the following method.

Fourth Reagents

04.01 Clarifying agent: basic lead acetate solution.

Into about 1000 ml of freshly boiled water was added 560 g dry
basic lead acetate. The mixture was boiled for 30 min and then allowed to stand overnight
.


Liquid layer was decanted and diluted with freshly boiled water to obtain a solution
a density of 1.25 g / ml (at 20 °. C). This solution must be
protect from air.

04.02 diethyl

Fifth Apparatus

05.01 Saccharimeter is calibrated to a normal weight of 26 g of sucrose, or polarimeter


This device must be placed in the room in which it is possible to maintain a temperature of around 20
st. C. The instrument was calibrated with standard quartz plates
.

05.02 Light source consisting of a sodium vapor lamp.

05.03 Precision polarimeter tubes with a length of 200 mm with a tolerance of +/- 0.02 mm
.

05.04 Analytical scales accurate to 0.1 mg.

05.05 Individually calibrated volumetric flask 100 ml stoppers.
Flasks with a real capacity in the range (100.00 +/- 0.01) ml can be used without correction
. Flasks outside this range may be
used only with proper correction to 100 ml.

06.05 Water bath with thermostat set at (20 +/- 0.1) st. C.

6th procedure

06.01 Preparation of the solution

ASAP Weigh (26 +/- 0.002) g and quantitatively
transferred into a volumetric flask of 100 ml (5.5) containing 60 ml
water.

Sample was dissolved agitation without heating.

If necessary clarified solution, 0.5 ml reagent
basic lead acetate (1/4).

Solution was stirred for swirling the flask and walls of the flask are rinsed
so long as the meniscus ascends about 10 mm below the calibration mark.

The flask was placed in a water bath (5.6) maintained at (20 +/- 0.1) st.
C and kept there until the temperature of the sugar solution
constant.

Any bubbles formed on the surface of the liquid drop is removed
diethyl ether (4/2).

Solution of water was added to the mark.

Stopper and mix thoroughly by inverting at least three times
upside down.

Flask was allowed to stand for five minutes.

06.02 Determination of polarization

During all of the following operations maintaining the temperature (20 +/- 0.1) st.
C.

6.2.1. Polarimeter is set to zero.

6.2.2. The sample was filtered through filter paper. The first 10 ml of filtrate
apply collected until another 50 ml of the filtrate.

6.2.3. Polarometrická tube is washed twice
rinsing with a solution of the sample to be tested (6.2.2.).

6.2.4. The tube is at a temperature (20 +/- 0.1) st. C carefully with
sample solution to be tested. When inserting the closure plate
to remove any air bubbles. The filled tube was placed into
cradle device.

6.2.5. Subtract the polarization to 0,05 st. S or
0.02 angular degrees. The same is repeated four more times. Calculate the average of these five measurements
.

7th Expression of results

07.01 Formula and method of calculation results are expressed in 0S
with an accuracy of 0.1 to Wed S. To convert the angular degrees 0S using the following formula:
st. S = degree angle x 2,889

07.02 repeatability

The difference between the results of two determinations carried out simultaneously or in rapid
themselves from the same sample by the same operator under the same conditions
not be greater than 0.1 st. S, the result of each determination is
average of five measurements.


Příl.23
Test methods for determining the purity of food additives

Introduction

First Preparation of the test sample

01.01 generally

Amount of laboratory sample to be tested must normally be 50 g
unless a determination is required for larger amounts

02.01 Sample Preparation

The sample must be homogenized before the test.

03.01 preservation

Prepared sample shall always be kept in an airtight and watertight
container and must be stored so as to prevent any damage.

Second Reagents

02.01 water

2.1.1. Wherever mention is made of water for solution, dilution or washing
mean distilled water or demineralized water of equivalent purity
.

2.1.2. Wherever there is mention of a solution or a dilution, without mentioning other
indication, means an aqueous solution.

02.02 Reagents Unless otherwise specified, all chemicals must be analytical grade
.

Third equipment

03.01 Device List

List of equipment contains only those items for special use and items to a particular specification
.

03.02 Analytical Balance


Analytical scales are scales with a sensitivity of 0.1 mg or higher.

Fourth Expression of results

04.01 results

The results presented in the analysis report shall be the average value
least two determinations, the repeatability is satisfactory.

04.02 Percent

Unless otherwise specified, the results must be expressed in weight percent
original sample as received at the laboratory.

04.03 The number of significant digits

Number of significant digits of the result so expressed shall be governed
precision of the method.


Příl.24
Methods for determination of substances extractable with diethyl ether from sulfonated
organic dyes soluble in water and for food

First Subject matter and use

The method set substances which can be extracted with diethyl ether
of sulphonated organic dyes, which have not been mixed with any
carrier.

Second definition

Ether extractables content means the content of substances determined by the method
.

Third Principle

Dyes Extraction with diethyl ether and weigh the extracted residue after evaporation of the ether
.

Fourth Reagents

04.01 Diethyl ether, without water, peroxide-free (dried freshly vyžíhaným
calcium chloride).

Fifth Apparatus

05.01 Soxhlet apparatus with flask.

05.02 Desiccator containing freshly activated silica gel or equivalent
hair with moisture indicator.

05.03 Analytical balance.

05.04 Oven, thermostatically controlled at 85 +/- 2 st. C.

6th procedure

On a piece of filter paper with 10 mg, weigh about 10 g
sample dye. The paper is folded, placed in paper
extraction cartridge which is closed with cotton wool fat free.
Extracted with diethyl ether (4.1) six hours in a Soxhlet extraction apparatus (5.1).
Ether is evaporated at the lowest possible temperature. Tared Soxhlet flask
along with the rest placed in an oven (5.4) and dried 20 min at
(85 +/- 2) st. C. The flask was placed in a desiccator (5.2)
loosely covered with a lid and allowed to cool. The flask and the residue is weighed.

Drying and weighing is repeated until two successive weighings differ by
less than 0.5 mg. If the weight of the flask increases will be used when calculating
lowest recorded value.

7th Expression of results

07.01 Formula and method of calculation

Ether extractables content expressed as a percentage of the sample is given by
:
m1 x 100
---------
m0
Where
:
m1 - the amount of the residue after evaporation (grams)
m0 - initial quantity of collected sample (g)

07.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions
not be greater than 20 mg per 100 g of sample.


Příl.25
Method of determining formic acid, formates and other oxidizable
impurities in acetic acid, potassium acetate, sodium diacetate and acetate
calcium

First Subject matter and use

The method determines formic acid, formates and other
oxidizable impurities, expressed as formic acid

- Acetic acid (E 260)

- In potassium acetate (E 261)

- In sodium diacetate (E 262)

- Of calcium acetate (E 263).

Second definition

The content of formic acid, formates and other oxidizable impurities
means the content of formic acid, formates and other oxidizable
impurities as determined by the method.

Third Principle

Treatment with excess potassium permanganate solution sample
alkaline environment is formed manganese dioxide. Manganese dioxide and
excess potassium permanganate are determined iodometrically in acidic medium
and concentration of oxidizable impurities is calculated and expressed as formic acid
.

Fourth Reagents

04.01 Potassium iodide.

04.02 Potassium permanganate: 0.02 mol / l.

04.03 Sodium carbonate (anhydrous).

04.04 Sodium thiosulfate 0.1 mol / l.

05.04 Starch solution (about 1% strength).

06.04 Dilute sulfuric acid: 90 ml of water was added 90 ml of sulfuric acid
(d20 = 1.84 g / ml).

Fifth Apparatus

05.01 Boiling water bath.

05.02 Analytical balance.

6th procedure

If the free acid sample, weighed to an accuracy of about 10 10 mg

G sample, diluted with 70 ml of water and a solution containing 10 g
anhydrous sodium carbonate (4.3) in 30 ml of water. If the sample is a salt,
weighed to an accuracy of 10 mg, about 10 g of sample and dissolved in 100 ml of water
. Add 1 g anhydrous sodium carbonate (4.3) and shake to dissolve
. Then 20 ml of potassium permanganate (4.2)
about 0.02 mol / l for 15 minutes was heated in a boiling water bath.
The mixture was cooled and 50 ml dilute sulfuric acid (4.6) and 0.5 g
potassium iodide (4.1). The mixture was stirred swirl until all
precipitated manganese dioxide dissolves again.
Titrated with sodium thiosulfate (4.4) at a concentration of 0.1 mol / l
to pale yellow color of the solution. Add a few drops of starch solution (4.5) and in
titration is continued until the solution is colorless.

7th Expression of results

07.01 Formula and method of calculation

Content of formic acid, formates and other oxidizable impurities
expressed in percentage as formic acid, is given by:
+ - - + 2,3 b
| 100a |
----- X | ------- - V |
m0 | b |
+ - - +
Where:
and - the molar concentration of potassium permanganate
b - the molar concentration of sodium thiosulfate
m0 - initial weight of sample (g)
V - volume of sodium thiosulphate 0,1 mol / l
consumed for the titration (ml).

07.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions
not be greater than 5 mg per 100 g of sample.

8th Notes

08.01 11.3 ml of sodium thiosulphate solution of 0,1 mol / l
is 0.2% formic acid in 10 g sample.

08.02 If there are no formates, the consumption of 20 ml, but
if present more than 0.27% by weight of formic acid,
excess potassium permanganate is not sufficient, and when the titration
consumes constant minimum volume of 8 ml. In this case, the determination
repeated with a smaller amount of sample.


Příl.26
Methods for determination of non-volatile substances in propionic acid

First Subject matter and use

The method determines non-volatile substances in propionic acid (E 280).

Second definition

The content of non-volatile substances in propionic acid content means
non-volatile substances determined by the method specified.

Third Principle

Sample is evaporated and then dried at (103 +/- 2) st. C and the residue
determined by weighing.

Fourth Apparatus

04.01 Evaporating dish porcelain or platinum of sufficient volume to accommodate
100 g of sample.

04.02 Oven, electrically heated, thermostatically controlled at (103 +/-
2) st. C.

04.03 Analytical balance.

04.04 Boiling water bath.

05.04 Desiccator containing freshly activated silica gel or equivalent desiccant
with moisture indicator.

Fifth procedure

Into a previously dried and weighed dish (4.1), weigh 100 g of sample
propionic acid with 0,1 grams. Evaporate on the boiling water bath
(4.4) in the hood. After evaporation of all propionic acid
dish is placed for one hour in the oven (4.2) at a temperature of (103 +/- 2)
Wed. C, then the desiccator and allow to cool and then weigh.
Heating, cooling and weighing is repeated until there is no difference between
two successive weighings is less than 0.5 mg. If the weight
increase will be used in calculating the lowest recorded value.

6th Expression of results

06.01 Formula and method of calculation

Non-volatile content, calculated as a percentage of the sample is given by:
100 x m1
---------
m0

Where:
m1 - mass of the residue after evaporation (grams)
M0 - the weight of sample (g).

06.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions
not be greater than 5 mg per 100 g of sample.


Příl.27
Method for determining the loss on drying with sodium nitrite

First Subject matter and use

The method determines the weight loss on drying with sodium nitrite (E
250).

Second definition

Moisture content of sodium nitrite refers to loss on drying
determined by the method specified.

Third Principle

Loss on drying is obtained by heating in an oven at

(103 +/- 2) st. C, weighing and calculation of weight loss.

Fourth Apparatus

04.01 Oven, electrically heated, thermostatically controlled at (103 +/-
2) st. C.

04.02 Weighing dish, flat-bottomed, glass, of diameter 60 to 80 mm and a depth
least 25 mm, with a loose lid.

04.03 Desiccator containing freshly activated silica gel or equivalent desiccant with
content indicator.

04.04 Analytical balance.

Fifth procedure

From the bowl on the weighing (4.2) removed the lid and bowl and lid hour
heated in the oven (4.1) at (103 +/- 2) st. C. bowl (4.2)
cover cap, placed in a desiccator (4.3) and allowed to cool to room temperature
. Covered bowl (4.2) was weighed to 10 mg
. To the bowl with a lid with an accuracy of 10 mg Weigh about 10 g of the sample.
Cap is removed and the dish (4.2) and lid is placed on one hour
oven (4.1) at (103 +/- 2) st. C. bowl is again covered with
lid and left in the desiccator (4.3) to cool to room temperature.
Weigh to the nearest 10 mg. Heating, cooling and weighing
repeated until the difference between two successive weighings is less than 10 mg
. If the weight increase will be used when calculating
lowest recorded value.

6th Expression of results

06.01 Formula and method of calculation

Loss on drying, calculated as a percentage of the weight of the sample is given by the formula
:
100 x (m2 - m3)
----------------
(M2 - m1)

Where:
m1 - weight tray (g)
m2 - weight of pan and sample before drying (g)
m3 - weight of pan and sample after drying (g).

06.02 repeatability

The difference between the results of two determinations carried out simultaneously or in rapid
themselves in the same sample by the same operator under the same conditions
not be greater than 100 mg per 100 g of sample.


Příl.28
Method for proof higher than the marginal amount of salicylic acid
ethyl 4-hydroxybenzoate sodium, ethyl 4-hydroxybenzoate sodium,
propyl 4-hydroxybenzoate, propyl 4-hydroxybenzoate sodium, methyl
-4-hydroxybenzoate and methyl 4-hydroxybenzoate sodium

First Subject matter and use

The method detects salicylic acid
ethyl 4-hydroxybenzoate (E 214), propyl 4-hydroxybenzoate (E 216) and
methyl 4-hydroxybenzoate (E 218) and their sodium salts (E 215, E 217, E 219
).

Second definition

Discovering the presence of salicylic acid in a concentration limit means
result of evidence of higher than the marginal amount determined by the method
.

Third Principle

Reaction of ferric ammonium sulphate with a solution of sample A violet coloration
. Its intensity is compared with the intensity of discoloration caused
reactions comparative solution.

Fourth Reagents

04.01 A solution of ferric ammonium sulphate, 0.2%: prepared by dissolving 0.2 g of dodecahydrate
ferric ammonium sulphate in 50 ml of water, add 10 ml
10 vol.% Nitric acid, and diluting with water to 100 ml .

04.02 Ethanol, 95 vol.%.

04.03 A solution of salicylic acid, 0.1 g / l.

04.04 Sulfuric acid, 1 mol / l.

Fifth Apparatus

05.01 Nessler cylinders with a resolution up to 50 ml with a total volume of approximately 60 ml
.

6th procedure

06.01 Samples ethyl, propyl, and methyl 4-hydroxybenzoate.

6.1.1. With the accuracy of 1 mg was weighed 0.1 g of sample and dissolved in 10 ml
95% ethanol (4.2). Transfer the solution to a graduated Nessler cylinder
(5.1) and diluted with water to 50 mL. Stir and under stirring 1 g
ferric ammonium sulphate (4.1). Allow a minute.

6.1.2. Simultaneously, repeat the procedure 6.1.1
prepared comparative solution, but 0.1 g of the sample was replaced with 1 ml of a solution of salicylic acid
(4.3).

6.1.3. Coloration of the sample solution is compared with the coloration comparative
solution.

06.02 Samples of the sodium salts of ethyl, propyl and methyl 4-hydroxybenzoate.

6.2.1. 6.1.1 Repeat the procedure, before dilution to 50 ml is made
acidification with sulfuric acid (4.4) at a concentration of 1 mol / l at pH 5

6.2.2. Repeat the procedure 6.1.2.

6.2.3. Repeat the procedure 6.1.3.

7th Expression of results

07.01 Evaluation of evidence overallotment amount

If a reddish purple color in the test tube with the sample solution
intense than the color of the tube with the comparative sample, the test
positive and the sample contains more than 0.1% salicylic acid.

07.02 sensitivity


Detection limit is 30 mg of salicylic acid per 100 g of sample.

07.03 Notes

The results of two evidence overallotment amount
carried out simultaneously or in rapid succession on the same sample and the same operator under the same conditions
must be the same.


Příl.29
Determination method of free acetic acid in sodium diacetate

First Scope and field of application The method determines acetic acid in
sodium diacetate (E 262).

Second definition

Acetic acid content means the content of acetic acid
determined by the method specified.

Third Principle

Direct titration of acetic acid in a sample of standard sodium hydroxide
solution using phenolphthalein as indicator.

Fourth Reagents

04.01 1% phenolphthalein solution in ethanol.

04.02 Sodium hydroxide, 1 mol / l.

Fifth Apparatus

05.01 Analytical balance.

6th procedure

Accuracy of 1 mg weigh about 3 g sample and dissolve in about 50 ml of water
. Add two or three drops of phenolphthalein solution (4.1) and
titrated with sodium hydroxide (4.2) at a concentration of 1 mol / l until the red
color lasts 5 sec.

7th Expression of results

07.01 Formula and method of calculation

Acetic acid content in percentage by mass of the sample is given by:
6005 x xc
--------------
m0

Where:
V - volume of the consumed sodium hydroxide (ml)
c - concentration of the NaOH solution (mol / l)
m0 - initial weight of sample (g).

07.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions
not be greater than 500 mg per 100 g of sample.

8th Note

For titration of 3 g of the sample containing 40% acetic acid is needed
20 ml of sodium hydroxide at a concentration of 1 mol / l.


Příl.30
Method for the determination of sodium acetate in sodium diacetate

First Subject matter and use

The method determines sodium acetate and water, expressed as sodium acetate
in sodium diacetate (E 262).

Second definition

Sodium acetate content means the content of sodium acetate and water
expressed as sodium acetate, determined by the method specified.

Third Principle

Sample is dissolved in glacial acetic acid and titrate with standard
perchloric acid using crystal violet as indicator

.

Fourth reagents

04.01 Glacial acetic acid (p20 = 1,049 g / ml) for anhydrous titration

04.02 Crystal violet (CI 42555), 0.2 wt. % Solution in glacial acetic acid
number.

04.03 Potassium hydrogen,

04.04 Acetic anhydride,

05.04 Perchloric acid 0,1 mol / l, in glacial acetic acid
. Must be prepared and standardized as follows: Do
1000-mL volumetric flask glass-stoppered Weigh P (g)
perchloric acid. The quantity P is calculated from the formula:
1004.6
P = -------
m

where
m is the concentration of perchloric acid in a weight
alkalimetric percentage determined by titration (most
concentration is 70-72 wt.%). Add about 100 ml
glacial acetic acid and then gradually in small doses
quantity Q (g) of acetic anhydride. During adding
the mixture is continually stirred and cooled. Q amount may
calculated from the formula:

(567 x P) - 5695
Q = -------------------
and

Where P is the weighed amount of perchloric acid and a is the concentration
acetic anhydride in wt. %. The flask was stoppered and allowed to stand for 24 hours
a dark place, then add a sufficient amount of glacial acetic acid
to yield 1000 ml.
The solution prepared in this way is practically anhydrous. The solution is standardized
hydrogenftalátem potassium follows:

With accuracy of 0.1 mg weigh about 0.2 grams of potassium hydrogen
previously dried for 2 hours at 110 st. C and in the titration flask under slight heating
dissolved in 25 ml of glacial acetic acid. Cool,
added two drops of 0.2% crystal violet solution (4.2) in glacial acetic acid
and titrate with perchloric acid until
color indicator changes to light green. Using the same volume of solvents
Carry out a blank titration and the blank value was subtracted from
obtained at the actual determination. Every 20.42 mg

Potassium hydrogen equivalent to 1 ml perchloric acid
0,1 mol / l.

Fifth Apparatus

05.01 Analytical balance.

6th procedure

Accuracy of 0.5 mg is weighed about 0.2 g and dissolved in 50 ml
glacial acetic acid (4.1). Add several drops of indicator
crystal violet solution (4.2) and titrate to a pale green coloration
standard solution of perchloric acid (4.5) 0,1
mol / l.

7th Expression of results

07.01 Formula and method of calculation

The content of sodium acetate, as defined in section 2 (definition), expressed as a percentage
weight of the sample is given by the following formula:
8,023 x V xc
---------------
m0

Where:
V - the volume of standard acid consumed
perchloric (4.5) (mL)
c - molar concentration of the perchloric acid solution (4.5)
M0 - the weight of sample (g).

07.02 repeatability

Difference between the results of two determinations, carried out simultaneously or in rapid succession
the same sample and the same operator on the same conditions
not be greater than 1.5 g per 100 g of sample.

8th Note

Reagents used in this method are toxic and require careful handling
.


Příl.31
Method for the detection of more than marginal quantities of aldehydes in acetic
sorbate in sodium sorbate, potassium and calcium and in propionic acid

First Subject matter and use

This method detects aldehydes, expressed as formaldehyde


- In sorbic acid (E 200)

- In sodium sorbate, potassium and calcium (E 201, E 202, E 203)

- In propionic acid (E 280).

Second definition

Discovering the presence of aldehydes in the threshold concentration means the outcome
evidence overallotment amount determined by the method specified.

Third Principle

Aldehydes in the test solution and a formaldehyde solution in Comparative
react with Schiff's reagent to produce red colored complexes
whose intensities are compared.

Fourth reagents

04.01 Formaldehyde solution (0.01 mg / ml) prepared by diluting
concentrated formaldehyde solution (400 mg / ml).

04.02 Schiff's reagent.

Fifth procedure

05.01 With the accuracy of 1 mg was weighed about 1 g of sample, add 100 ml of water and shake
. If necessary, the solution is filtered and 1 ml of filtrate
or sample, add 1 ml of Schiff's reagent (4.2). Simultaneously to 1 ml
Comparative formaldehyde solution was added 1 ml of Schiff's reagent (4.2).

05.02 Coloration of the sample solution is compared with the coloration comparative
solution.

6th Expression of results

06.01 Evaluation evidence overallotted quantity If
red coloration in the test tube with the sample solution in intense coloration than
tube comparative solution, the test is positive and the sample
contains more than 0.1% aldehydes, expressed as formaldehyde.

06.02 sensitivity

Limit of detection of this test is 30 mg of formaldehyde per 100 g of sample.

06.03 Notes

Results evidence overallotted two quantities that are carried
simultaneously or in rapid succession on the same sample, under the same
same conditions must be the same.


Příl.32
Method of determining the peroxide number of lecithins

First Subject matter and use

This method determines the peroxide number in lecithins (E 322).

Second definition

Perhydrate number of lecithins means the outcome determined by the method
.

Third Principle

Oxidation of potassium iodide peroxides of lecithin and titration of the liberated iodine
standard sodium thiosulphate.

Fourth Reagents

04.01 Glacial acetic acid.

04.02 Chloroform.

04.03 Potassium iodide.

04.04 Sodium thiosulfate 0.1 mol / l or 0.01 mol / l.

05.04 Starch solution (about 1%).

Fifth Apparatus

05.01 Analytical balance.

05.02 Apparatus, see the picture, which consists of:

5.2.1. Ground-glass 100 ml round-bottomed flask;

5.2.2. reflux condenser;

5.2.3. glass tube, 250 mm long with an internal diameter of 22 mm,
ground joint;

Mikrokádinky external dimensions - 20 mm diameter and height 35-50 mm.

6th procedures

06.01 In a 100 mL flask (4.1) was poured 10 ml of glacial acetic acid (4.1)
and 10 ml of chloroform (4.2). Fit the glass tube (5.2.3) and re
condenser (5.2.2) and the mixture gently boiled for two minutes to expel all

Dissolved air. 1 g of potassium iodide (4.3) dissolved in 1.3 ml
water and this solution was added to the mixture in the flask (5.2.1), while care is taken
not to interrupt boiling.

If at this stage appear yellow color determination must be annulled
and must be repeated with fresh reagents.

06.02 With an accuracy of 1 mg weigh about 1 g of sample and after a further two minutes of boiling
weighed sample is added to the contents of the flask (5.2.1) again
must take care not to interrupt var. Therefore the sample is placed in
mikrokádince (5.2.4) with suitably shaped bottom, as shown in Scheme
which can be triggered glass tube with a glass rod
(5.2.3). Condenser (5.2.2) may be removed for a short time.
At reflux was continued for three to four minutes. The heat was
ends and immediately disconnect the condenser (5.2.2). Glass tube (5.2.3)
rapidly added 50 ml of water. The glass tube (5.2.3) is removed and the flask (5.2.1)
under water supply was cooled to room temperature. Titrate with sodium thiosulphate
(0.1 mol / l or 0.01 mol / l) (4.4) until the aqueous layer
changed color to light yellow. Add 1 ml of the starch solution (4.5) and titrating
is continued until the blue color. Flask (5.2.1) during titration
thoroughly shaken to ensure complete extraction of iodine from
nonaqueous layer.

06.03 Blind titration value was obtained by repeating the whole procedure
6.1 and 6.2, but without adding the sample.

7th Expression of results

07.01 Formula and method of calculation The peroxide number in the sample
milliequivalents per kilogram is given by the formula:
1000 xax (V1 - V2)
--------------------
m0

Where:
V1 - volume of sodium thiosulfate consumed in the titration
sample (6.2) (mL)
V2 - volume of sodium thiosulfate consumed in the titration
the blank (6.3) (ml)
and - the concentration of the sodium thiosulphate solution (mol / l)
m0 - initial weight of sample (g).

07.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions has
be greater than 0.5 (expressed as a peroxide number in milliequivalents per kilogram of sample
) .

8th Notes

08.01 The choice of concentration of sodium thiosulphate
depends on the expected outcome titration. When using less than 0.5 ml
sodium thiosulphate 0,1 mol / l, repeated
assay using sodium thiosulfate
0,01 mol / l.

08.02 The determination should not be carried out in strong light.
Graphical representation of apparatus for determining
peroxide number in lecithins


Příl.33
The method of determination of substances insoluble in toluene contained in lecithin

First Subject matter and use

The method determines the toluene-insoluble substances contained in
lecithin (E322).

Second definition

Insoluble content in toluene means
content of substances insoluble in toluene determined by the method specified.

Third Principle A sample is dissolved in toluene, filtered and the residue
dried and weighed.

Fourth Reagents

04.01 toluene

Fifth Apparatus

05.01 Crucible fritted funnel volume of 30 ml, a porosity G3 or equivalent.

05.02 Drying oven, electrically heated and thermostatically controlled at (103 +/-
2) st. C.

05.03 Water bath, operating at a temperature not exceeding 60 st. C.

05.04 Desiccator containing freshly activated silica gel or equivalent desiccant with
content indicator.

05.05 500 ml conical flask.

06.05 Vacuum pump.

07.05 Analytical balance.

6th procedure

06.01 Cup of a 30 ml sintered glass (5.1) is dried in an oven at (103
+/- 2) st. C (5.2). The crucible was transferred to a desiccator (5.4), allow to cool
and then weighed.

06.02 Sample lecithins after optional heating in a water bath (5.3)
thoroughly. The conical flask (5.5) with an accuracy of 1 mg
carefully weigh about 10 grams of sample. Add 100 ml of toluene (4.1) and the mixture was stirred
swirl until all lecithin
apparently dissolved. The solution is filtered through a fritted crucible (5.1).
Conical flask (5.5) is rinsed with 25 ml of toluene (4.1) and the washings shed
crucible (5.1). This procedure was repeated with another 25 ml of toluene (4.1).
Excess toluene from the crucible (5.1) is removed by suction.


06.03 Crucible (5.1) is dried in an oven (5.2) for two hours at (103 +/- 2)
Wed. C. Place in desiccator (5.4) and allow to cool.
After cooling, the crucible is weighed rest.

06.04 6.3 The process is repeated until the difference between two consecutive
successive weighings is less than 0.5 mg. If the weight increases
used in calculating the lowest recorded value.

7th Expression of results

07.01 Formula and method of calculation insoluble content in toluene is given by the formula
:
100 (m2 - m1)
--------------
m0

Where:
m1 - mass of the empty crucible (6.1) (g)
m2 - the weight of the crucible and the residue (6.4) (g)
M0 - the weight of sample (g).

07.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions has
be greater than 30 mg per 100 g of sample.


Příl.34
Method for the detection of higher than the marginal amount
reducing substances in sodium lactate, potassium and calcium

First Subject matter and use

Test is used for the qualitative evidence of reducing substances

- In sodium lactate (E 325)

- In potassium lactate (E 326)

- In calcium lactate (E 327).

Second definition

Proof of reducing substances means the outcome of an overallotment evidence
amount determined by the method specified.

Third Principle

Fehling's solution is reduced by compounds having reducing ability. Such substances are usually
reducing sugars.

Fourth Reagents

04.01 Fehling's solution A: 6.93 g of copper sulfate pentahydrate was
dissolved in water and make up to 100 ml with water.

04.02 Fehling's solution B: 34.6 g potassium sodium tartrate and 10 g
sodium hydroxide were dissolved in water and made up to 100 ml with water
.

Fifth procedures

Accuracy of 1 mg weigh about 1 g of sample and dissolved in 10 ml of warm water
. Add 2 ml of Fehling's solution A (4.1) and 2 ml of Fehling's solution
B (4.2), then the mixture was boiled for one minute and it is observed whether
will change color. Calcium sulfate, which is sometimes precipitated
determination affected.

6th Expression of results

06.01 Evaluation of evidence overallotment amount

If after boiling (5) changes the color of the test is positive and
presence of reducing agents is demonstrated.

06.02 sensitivity

Between the detection of reducing substances is 100 mg glucose per 100 g of sample.

06.03 Notes

6.3.1. The results of two evidence overallotted amount
carried out simultaneously or in rapid succession on the same sample and the same operator on the same
conditions must be the same.

6.3.2. Both Fehling solutions react if they are present in the sample
2% glucose.


Příl.35
The method of determination of volatile acids in orthophosphoric acid

First Subject matter and use

The method determines volatile acids, expressed as acid
acid, in orthophosphoric acid (E 338).

Second definition

Volatile acid content means the content of volatile acids, expressed as acetic acid
determined by the method specified.

Third Principle

The sample water was added and the solution was distilled. The distillate is titrated
standard sodium hydroxide solution. Acidity is expressed as
acetic acid.

Fourth Reagents

04.01 1% phenolphthalein solution in ethanol.

04.02 Sodium hydroxide, 0.01 mol / l.

Fifth Apparatus

05.01 Distillation apparatus with droplet separator.

6th procedure

Accuracy of 50 mg was weighed about 60 g of the sample and the sample weighed, and 75
ml of freshly boiled and cooled water is introduced into a distillation flask fitted
droplet separator (5.1). Mix and then distil
about 50 ml solution.

The distillate is titrated against standard sodium hydroxide solution (4.2)
a concentration of 0.01 mol / l, using phenolphthalein (4.1) as indicator.
Titration is continued until the first red color of the solution persists for 10
s.

7th Expression of results

07.01 Formula and method of calculation

The content of volatile acids, expressed as milligrams per kilogram of acetic acid
, is given by:
600 x
---------
m0

Where:
V - the volume of sodium hydroxide 0,01 mol / l
consumed for the titration (ml)
m0 - sample weight orthophosphoric acid (g).

07.02 repeatability


Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions
not exceed 1 mg per 100 g of sample.


Příl.36
Method for the detection of higher than the marginal amount of nitrate in phosphoric acid


First Subject matter and use

This method detects the presence of nitrates in orthophosphoric acid
(E 338).

Second definition

Proof of the presence of nitrate, expressed as sodium nitrate is
means the outcome of an overallotment amount of evidence set
prescribed method.

Third Principle

Sample was added to a solution of indigo carmine in a concentrated sulfuric acid
. Original blue discoloration disappear under oxidizing
substances, including nitrates.

Fourth Reagents

04.01 Indigo carmine solution, 0.18%: 0.18 g of sodium indigotinsulfonátu
was dissolved in water and make up to 100 ml with water.

04.02 Solution of sodium chloride, 0.05%.

04.03 Concentrated sulfuric acid (20 = 1,84 g / ml).

Fifth procedure

Measure 2 ml of sample and dilute sodium chloride solution (4.2) to 10 ml
. Add 0.1 ml of a solution of indigo carmine (4.1) and then slowly
added 10 ml of concentrated sulfuric acid (4.3), during the addition
cooled. It is observed that the blue color of the solution persists for five minutes.

6th Expression of results

06.01 Evaluation of evidence overallotment amount

If the blue coloration disappears within five minutes, the test is positive and
content of oxidizing agents, expressed as sodium nitrate, is greater than
5 mg / kg.

06.02 Notes

6.2.1. Carry out the blank.

6.2.2. The results of two evidence overallotted quantity
carried out simultaneously or in rapid succession on the same sample and the same operator on the same
conditions must be the same.

6.2.3. Indigo carmine solution is older than 60 days should not be used.

6.2.4. If a positive result is obtained the sample may contain
nitrates and other oxidizing agents and the test must be repeated by
method ISO 3709 (1976), "Phosphoric acid for industrial use
(including foodstuffs) - determination of oxides of nitrogen
spectrophotometric method with 3,4-xylenol. "


Příl.37
Methods for determination of water-insoluble substances present in
sodium orthophosphate, di- and tri-orthophosphate and potassium
dipotassium and tripotassium

First Subject matter and use

The method determines water insoluble

- In sodium orthophosphate (E 339a)

- In Disodium orthophosphate (E 339b)

- In orthophosphate tri (E 339 c)

- In potassium orthophosphate (E 340 a)

- In dipotassium orthophosphate (E 340b)

- In tripotassium orthophosphate (E 340c).

Second definition

Substances insoluble in water means the content of substances insoluble in water
determined by the method specified.

Third Principle

Sample is dissolved in water and filtered through a suitable porcelain crucible.
After washing and drying the residue is weighed and calculated as content of substances
insoluble in water.

Fourth Apparatus

04.01 Crucible with frit, porosity G3 or equivalent.

04.02 Desiccator containing freshly activated silica gel with humidity indicator
or equivalent desiccant with a humidity indicator.

04.03 Drying thermostatically controlled at (103 +/- 2) st. C.

04.04 400 ml polypropylene beaker.

05.04 Boiling water bath.

Fifth procedure

S 10 mg, weigh about 10 g of sample in a beaker of phosphate and
(4.4) are dissolved in 100 ml of hot water by bringing to the boil for 15 minutes and
heating on a water bath (4.5). The solution is filtered, cleaned, dried and weighed
crucible (4.1). The insoluble residue was washed with hot water
. The crucible with residue in an oven (4.3) and two
hours drying at (103 +/- 2) st. C.

Place the crucible in a desiccator, allow to cool and then weigh.
Drying, cooling and weighing is repeated until the difference between two successive
weighing less than 0.5 mg. On increasing the weight, it is used in the calculation
lowest recorded value.

6th Expression of results

06.01 Formula and method of calculation

The content of water-insoluble substances in% by weight is given by:
m1
- 100 x
m0

Where:
m1 - mass of the residue after drying (g)

M0 - the weight of sample (g).

06.02 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator on the same conditions
not be greater than 10 mg per 100 g of sample.


Příl.38
Methods determine the pH of food additives

First Subject matter and use

In this method are given general guidelines for the determination of pH
food additives.

Second definition

The pH value of food additives means pH
determined by the method specified.

Third Principle

PH of the aqueous solution of the dissolved or slurried sample
determined in conventional manner using a glass electrode, reference electrode and
pH meter.

Fourth reagents

04.01 The instrument is calibrated using the following buffer solutions:

4.1.1. A buffer solution which has at 20 Wed. C, pH 6.88, consisting of equal volumes
potassium dihydrogen phosphate concentration
0.05 mol / l dibasic sodium phosphate at a concentration of 0.05 mol / l.

4.1.2. A buffer solution which has at 20 Wed. C pH 4, the solution of potassium hydrogen
0,05 mol / l.

4.1.3. A buffer solution which has at 20 Wed. C pH 9 solution is sodium borate solution
0,05 mol / l.

04.02 Saturated solution of sodium or potassium chloride
3 mol / l, or other suitable solution prescribed by the manufacturer of the electrode, to fill
reference electrode.

04.03 Distilled water without carbon dioxide, having a pH of 5 to 6

Fifth Apparatus

05.01 pH meter with an accuracy of 0.01 pH units.

05.02 Electrodes combined glass electrode or single
glass electrode and a reference electrode with suitable clamps to attach
.

05.03 Magnetic stirrer and heating element.

05.04 Thermometer, calibrated in the range 0-100 Wed. C.

6th procedure

06.01 Calibration of pH meter

Glass electrodes are mounted according to the manufacturer's instructions. PH
obtained by electrodes must be regularly checked by comparison with buffer
solutions of known pH.

Before insertion into the sample solution / buffer solution, the electrodes
rinsed with water and then gently wiped with a soft cloth or rinsed
water and then twice with the sample solution or calibration solution.
If the sample has a pH in the acidic range, take for checking
pH buffer solutions of pH 4 (4.1.2) and pH 6.88 (4.1.1). If the test sample
pH value in the alkaline range, are used for the control values ​​
pH buffer solutions of pH 9.22 (4.1.3) and pH 6.88 (4.1.1).

06.02 Measurement of the sample solution

Concentration of the sample used or the process used sample preparation
prescribed in the applicable regulations of the European Communities for
food additives.

Sample solution was prepared according to the instructions using distilled water
(4.3) and then stirring the temperature adjusted to 20 st. C. Agitation was discontinued
, to the solution is inserted glass electrode and after two minutes
subtracted pH (5.1).

7th Expression of results

07.01 repeatability

Difference between the results of two determinations carried out simultaneously or in rapid succession on
same sample and the same operator under the same conditions must be
greater than 0.05 pH unit.

8th Note

This method is applicable only when the regulations of the European
Community rules on food additives set
requirements for the pH of food additives
dissolved or suspended in water.


Příl.39 Reference tables


The refractive indices (n) sucrose solutions at 20 Wed. C ^ 1)

+-------------------+-----------------+-------------------+-------------------+--------------------+
| n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose |
| Wed C) (%) | st. C) (%) | st. C) ( %) | Wed C) (%) | Wed C) (%) |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| 1.3330 0.009 | 2.436 1.3365 | 1.3400 4821 | 1.3435 7164 | 1.3470 9466 |
| 1.3331 0.078 | 2.505 1.3366 | 1.3401 4888 | 1.3436 7230 | 1.3471 9531 |
| 1.3332 0.149 | 2.574 1.3367 | 1.3402 4956 | 1.3437 7296 | 1.3472 9596 |
| 1.3333 0.218 | 2.642 1.3368 | 1.3403 5023 | 1.3438 7362 | 1.3473 9661 |

| 1.3334 0.288 | 2.711 1.3369 | 1.3404 5091 | 1.3439 7429 | 1.3474 9726 |
| 1.3335 0.358 | 2.779 1.3370 | 1.3405 5158 | 1.3440 7495 | 1.3475 9791 |
| 1.3336 0.428 | 2.848 1.3371 | 1.3406 5225 | 1.3441 7561 | 1.3476 9856 |
| 1.3337 0.498 | 2.917 1.3372 | 1.3407 5293 | 1.3442 7627 | 1.3477 9921 |
| 1.3338 0.567 | 2.985 1.3373 | 1.3408 5360 | 1.3443 7693 | 1.3478 9986 |
| 1.3339 0.637 | 3.053 1.3374 | 1.3409 5427 | 1.3444 7759 | 1.3479 10.051 |
| 1.3340 0.707 | 3.122 1.3375 | 1.3410 5494 | 1.3445 7825 | 1.3480 10.116 |
| 1.3341 0.776 | 3.190 1.3376 | 1.3411 5562 | 1.3446 7891 | 1.3481 10.181 |
| 1.3342 0.846 | 3.259 1.3377 | 1.3412 5629 | 1.3447 7957 | 1.3482 10.246 |
| 1.3343 0.915 | 3.327 1.3378 | 1.3413 5696 | 1.3448 8023 | 1.3483 10.311 |
| 1.3344 0.985 | 3.395 1.3379 | 1.3414 5763 | 1.3449 8089 | 1.3484 10.375 |
| 1.3345 1.054 | 3.463 1.3380 | 1.3415 5830 | 1.3450 8155 | 1.3485 10.440 |
| 1.3346 1.124 | 3.532 1.3381 | 1.3416 5897 | 1.3451 8221 | 1.3486 10.505 |
| 1.3347 1.193 | 3.600 1.3382 | 1.3417 5964 | 1.3452 8287 | 1.3487 10.570 |
| 1263 1.3348 | 1.3383 3668 | 1.3418 6031 | 1.3453 8352 | 1.3488 10.634 |
| 1.3349 1.332 | 3.736 1.3384 | 1.3419 6098 | 1.3454 8418 | 1.3489 10.699 |
| 1.3350 1.401 | 3.804 1.3385 | 1.3420 6165 | 1.3455 8484 | 1.3490 10.763 |
| 1470 1.3351 | 1.3386 3872 | 1.3421 6231 | 1.3456 8550 | 1.3491 10.828 |
| 1540 1.3352 | 1.3387 3940 | 1.3422 6298 | 1.3457 8615 | 1.3492 10.892 |
| 1.3353 1.609 | 4.008 1.3388 | 1.3423 6365 | 1.3458 8681 | 1.3493 10,957 |
| 1678 1.3354 | 1.3389 4076 | 1.3424 6432 | 1.3459 8746 | 1.3494 11.021 |
| 1.3355 1.747 | 4.144 1.3390 | 1.3425 6498 | 1.3460 8812 | 1.3495 11.086 |
| 1.3356 1.816 | 4.212 1.3391 | 1.3426 6565 | 1.3461 8878 | 1.3496 11.150 |
| 1885 1.3357 | 1.3392 4279 | 1.3427 6.632 | 1.3462 8943 | 1.3497 11.215 |
| 1.3358 1.954 | 4.347 1.3393 | 1.3428 6698 | 1.3463 9008 | 1.3498 11.279 |
| 1.3359 2.023 | 4.415 1.3394 | 1.3429 6765 | 1.3464 9074 | 1.3499 11.343 |
| 1.3360 2.092 | 4.483 1.3395 | 1.3430 6831 | 1.3465 9139 | 1.3500 11.407 |
| 2161 1.3361 | 1.3396 4550 | 1.3431 6898 | 1.3466 9205 | 1.3501 11.472 |
| 2230 1.3362 | 1.3397 4618 | 1.3432 6.964 | 1.3467 9270 | 1.3502 11.536 |
| 1.3363 2.299 | 4.686 1.3398 | 1.3433 7031 | 1.3468 9335 | 1.3503 11.600 |
| 1.3364 2.367 | 4.753 1.3399 | 1.3434 7097 | 1.3469 9400 | 1.3504 11.664 |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose |
| Wed C) (%) | st. C) (%) | st. C) ( %) | Wed C) (%) | Wed C) (%) |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| 1.3505 11.728 | 15.207 1.3560 | 1.3615 18.595 | 1.3670 21.896 | 1.3725 25.114 |
| 1.3506 11.792 | 15.269 1.3561 | 1.3616 18.655 | 1.3671 21.955 | 1.3726 25.172 |
| 1.3507 11.856 | 15.332 1.3562 | 1.3617 18.716 | 1.3672 22.014 | 1.3727 25.230 |
| 1.3508 11.920 | 15.394 1.3563 | 1.3618 18.777 | 1.3673 22.073 | 1.3728 25.287 |
| 1.3509 11.984 | 15.456 1.3564 | 1.3619 18.837 | 1.3674 22.132 | 1.3729 25.345 |
| 1.3510 12.048 | 15.518 1.3565 | 1.3620 18.898 | 1.3675 22.192 | 1.3730 25.403 |
| 1.3511 12.112 | 15.581 1.3566 | 1.3621 18.959 | 1.3676 22.251 | 1.3731 25.460 |
| 1.3512 12.176 | 15.643 1.3567 | 1.3622 19.019 | 1.3677 22.310 | 1.3732 25.518 |
| 1.3513 12.240 | 15.705 1.3568 | 1.3623 19.080 | 1.3678 22.369 | 1.3733 25.576 |
| 1.3514 12.304 | 15.767 1.3569 | 1.3624 19.141 | 1.3679 22.428 | 1.3734 25.633 |
| 1.3515 12.368 | 15.829 1.3570 | 1.3625 19.201 | 1.3680 22.487 | 1.3735 25.691 |
| 1.3516 12.431 | 15.891 1.3571 | 1.3626 19.262 | 1.3681 22.546 | 1.3736 25.748 |
| 1.3517 12.495 | 15.953 1.3572 | 1.3627 19.322 | 1.3682 22.605 | 1.3737 25.806 |
| 1.3518 12.559 | 16.016 1.3573 | 1.3628 19.382 | 1.3683 22.664 | 1.3738 25.863 |
| 1.3519 12.623 | 16.078 1.3574 | 1.3629 19.443 | 1.3684 22.723 | 1.3739 25.921 |
| 1.3520 12.686 | 16.140 1.3575 | 1.3630 19.503 | 1.3685 22.781 | 1.3740 25.978 |
| 1.3521 12.750 | 16.201 1.3576 | 1.3631 19.564 | 1.3686 22.840 | 1.3741 26.035 |
| 1.3522 12.813 | 16.263 1.3577 | 1.3632 19.624 | 1.3687 22.899 | 1.3742 26.093 |
| 1.3523 12.877 | 16.325 1.3578 | 1.3633 19.684 | 1.3688 22.958 | 1.3743 26.150 |

| 1.3524 12.940 | 16.387 1.3579 | 1.3634 19.745 | 1.3689 23.017 | 1.3744 26.207 |
| 1.3525 13.004 | 16.449 1.3580 | 1.3635 19.805 | 1.3690 23.075 | 1.3745 26.265 |
| 1.3526 13.067 | 16.511 1.3581 | 1.3636 19.865 | 1.3691 23.134 | 1.3746 26.322 |
| 1.3527 13.131 | 16.573 1.3582 | 1.3637 19.925 | 1.3692 23.193 | 1.3747 26.379 |
| 1.3528 13.194 | 16.634 1.3583 | 1.3638 19.985 | 1.3693 23.251 | 1.3748 26.436 |
| 1.3529 13.258 | 16.696 1.3584 | 1.3639 20.045 | 1.3694 23.310 | 1.3749 26.493 |
| 1.3530 13.321 | 16.758 1.3585 | 1.3640 20.106 | 1.3695 23.369 | 1.3750 26.551 |
| 1.3531 13.384 | 16.819 1.3586 | 1.3641 20.166 | 1.3696 23.427 | 1.3751 26.608 |
| 1.3532 13.448 | 16.881 1.3587 | 1.3642 20.226 | 1.3697 23.486 | 1.3752 26.665 |
| 1.3533 13.511 | 16.943 1.3588 | 1.3643 20.286 | 1.3698 23.544 | 1.3753 26.722 |
| 1.3534 13.574 | 17.004 1.3589 | 1.3644 20.346 | 1.3699 23.603 | 1.3754 26.779 |
| 1.3535 13.637 | 17.066 1.3590 | 1.3645 20.406 | 1.3700 23.661 | 1.3755 26.836 |
| 1.3536 13.700 | 17.127 1.3591 | 1.3646 20.466 | 1.3701 23.720 | 1.3756 26.893 |
| 1.3537 13.763 | 17.189 1.3592 | 1.3647 20.525 | 1.3702 23.778 | 1.3757 26.950 |
| 1.3538 13.826 | 17.250 1.3593 | 1.3648 20.585 | 1.3703 23.836 | 1.3758 27.007 |
| 1.3539 13.890 | 17.311 1.3594 | 1.3649 20.645 | 1.3704 23.895 | 1.3759 27.064 |
| 1.3540 13.953 | 17.373 1.3595 | 1.3650 20.705 | 1.3705 23.953 | 1.3760 27.121 |
| 1.3541 14.016 | 17.434 1.3596 | 1.3651 20.765 | 1.3706 24.011 | 1.3761 27.178 |
| 1.3542 14.079 | 17.496 1.3597 | 1.3652 20.825 | 1.3707 24.070 | 1.3762 27.234 |
| 1.3543 14.141 | 17.557 1.3598 | 1.3653 20.884 | 1.3708 24.128 | 1.3763 ​​27.291 |
| 1.3544 14.204 | 17.618 1.3599 | 1.3654 20.944 | 1.3709 24.186 | 1.3764 27.348 |
| 1.3545 14.267 | 17.679 1.3600 | 1.3655 21.004 | 1.3710 24.244 | 1.3765 27.405 |
| 1.3546 14.330 | 17.741 1.3601 | 1.3656 21,063 | 1.3711 24.302 | 1.3766 27.462 |
| 1.3547 14.393 | 17.802 1.3602 | 1.3657 21.123 | 1.3712 24.361 | 1.3767 27.518 |
| 1.3548 14.456 | 17.863 1.3603 | 1.3658 21.183 | 1.3713 24.419 | 1.3768 27.575 |
| 1.3549 14.518 | 17.924 1.3604 | 1.3659 21.242 | 1.3714 24.477 | 1.3769 27.632 |
| 14581 1.3550 | 1.3605 17,985 | 1.3660 21.302 | 1.3715 24.535 | 1.3770 27.688 |
| 1.3551 14.644 | 18.046 1.3606 | 1.3661 21.361 | 1.3716 24.593 | 1.3771 27.745 |
| 1.3552 14.707 | 18.107 1.3607 | 1.3662 21.421 | 1.3717 24.651 | 1.3772 27.802 |
| 1.3553 14.769 | 18.168 1.3608 | 1.3663 21.480 | 1.3718 24.709 | 1.3773 27.858 |
| 1.3554 14.832 | 18.229 1.3609 | 1.3664 21.540 | 1.3719 24,767 | 1.3774 27.915 |
| 1.3555 14.894 | 18.290 1.3610 | 1.3665 21.599 | 1.3720 24.825 | 1.3775 27.971 |
| 1.3556 14.957 | 18.351 1.3611 | 1.3666 21.658 | 1.3721 24.883 | 1.3776 28.028 |
| 1.3557 15.019 | 18.412 1.3612 | 1.3667 21.718 | 1.3722 24.941 | 1.3777 28.084 |
| 1.3558 15.082 | 18.473 1.3613 | 1.3668 21.777 | 1.3723 24.998 | 1.3778 28.141 |
| 1.3559 15.144 | 18.534 1.3614 | 1.3669 21.836 | 1.3724 25.056 | 1.3779 28.197 |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose |
| Wed C) (%) | st. C) (%) | st. C) ( %) | Wed C) (%) | Wed C) (%) |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| 1.3780 28.253 | 31.317 1.3835 | 1.3890 34.310 | 1.3945 37.233 | 1.4000 40.091 |
| 1.3781 28.310 | 31.372 1.3836 | 1.3891 34.363 | 1.3946 37.286 | 1.4001 40.142 |
| 1.3782 28.366 | 31.428 1.3837 | 1.3892 34.417 | 1.3947 37.338 | 1.4002 40.194 |
| 1.3783 28.422 | 31.482 1.3838 | 1.3893 34.471 | 1.3948 37.391 | 1.4003 40.245 |
| 28479 1.3784 | 1.3839 31,537 | 1.3894 34.524 | 1.3949 37.443 | 1.4004 40.296 |
| 1.3785 28.535 | 31.592 1.3840 | 1.3895 34.578 | 1.3950 37.495 | 1.4005 40.348 |
| 1.3786 28.591 | 31.647 1.3841 | 1.3896 34.632 | 1.3951 37.548 | 1.4006 40.399 |
| 1.3787 28.648 | 31.702 1.3842 | 1.3897 34.685 | 1.3952 37.600 | 1.4007 40.450 |
| 1.3788 28.704 | 31.757 1.3843 | 1.3898 34.739 | 1.3953 37.653 | 1.4008 40.501 |
| 1.3789 28.760 | 31.812 1.3844 | 1.3899 34.793 | 1.3954 37.705 | 1.4009 40.553 |
| 1.3790 28.816 | 31.867 1.3845 | 1.3900 34.846 | 1.3955 37.757 | 1.4010 40.604 |
| 1.3791 28.872 | 31.922 1.3846 | 1.3901 34.900 | 1.3956 37.810 | 1.4011 40.655 |
| 1.3792 28.928 | 31.976 1.3847 | 1.3902 34.953 | 1.3957 37.862 | 1.4012 40.706 |

| 1.3793 28.984 | 32.031 1.3848 | 1.3903 35.007 | 1.3958 37.914 | 1.4013 40.757 |
| 1.3794 29.040 | 32.086 1.3849 | 1.3904 35.060 | 1.3959 37.967 | 1.4014 40.808 |
| 1.3795 29.096 | 32.140 1.3850 | 1.3905 35.114 | 1.3960 38.019 | 1.4015 40.860 |
| 1.3796 29.152 | 32.195 1.3851 | 1.3906 35,167 | 1.3961 38.071 | 1.4016 40.911 |
| 1.3797 29.208 | 32.250 1.3852 | 1.3907 35.220 | 1.3962 38.123 | 1.4017 40.962 |
| 1.3798 29.264 | 32.304 1.3853 | 1.3908 35.274 | 1.3963 38.175 | 1.4018 41.013 |
| 1.3799 29.320 | 32.359 1.3854 | 1.3909 35.327 | 1.3964 38.228 | 1.4019 41.064 |
| 1.3800 29.376 | 32.414 1.3855 | 1.3910 35.380 | 1.3965 38.280 | 1.4020 41.115 |
| 1.3801 29.432 | 32.468 1.3856 | 1.3911 35,434 | 1.3966 38.332 | 1.4021 41.166 |
| 1.3802 29.488 | 32.523 1.3857 | 1.3912 35.487 | 1.3967 38.384 | 1.4022 41.217 |
| 1.3803 29.544 | 32.577 1.3858 | 1.3913 35.540 | 1.3968 38.436 | 1.4023 41.268 |
| 1.3804 29.600 | 32.632 1.3859 | 1.3914 35.593 | 1.3969 38.488 | 1.4024 41.318 |
| 1.3805 29.655 | 32.686 1.3860 | 1.3915 35.647 | 1.3970 38.540 | 1.4025 41.369 |
| 1.3806 29.711 | 32.741 1.3861 | 1.3916 35,700 | 1.3971 38.592 | 1.4026 41.420 |
| 1.3807 29.767 | 32.795 1.3862 | 1.3917 35.753 | 1.3972 38.644 | 1.4027 41.471 |
| 1.3808 29.823 | 32.849 1.3863 | 1.3518 35.806 | 1.3973 38.696 | 1.4028 41.522 |
| 1.3809 29.878 | 32.904 1.3864 | 1.3919 35.859 | 1.3974 38.748 | 1.4029 41.573 |
| 1.3810 29.934 | 32.958 1.3865 | 1.3920 35.912 | 1.3975 38.800 | 1.4030 41.623 |
| 1.3811 29.989 | 33.013 1.3866 | 1.3921 35.966 | 1.3976 38.852 | 1.4031 41.674 |
| 1.3812 30.045 | 33.067 1.3867 | 1.3922 36.019 | 1.3977 38.904 | 1.4032 41.725 |
| 1.3813 30.101 | 33.121 1.3868 | 1.3923 36.072 | 1.3978 38.955 | 1.4033 41.776 |
| 1.3814 30.156 | 33.175 1.3869 | 1.3924 36.125 | 1.3979 39.007 | 1.4034 41.826 |
| 1.3815 30.212 | 33.230 1.3870 | 1.3925 36.178 | 1.3980 39.059 | 1.4035 41.877 |
| 1.3816 30,267 | 33,284 1.3871 | 1.3926 36.231 | 1.3981 39.111 | 1.4036 41.928 |
| 1.3817 30.323 | 33.338 1.3872 | 1.3927 36.284 | 1.3982 39.163 | 1.4037 41.978 |
| 1.3818 30.378 | 33.392 1.3873 | 1.3928 36.337 | 1.3983 39.214 | 1.4038 42.029 |
| 1.3819 30,434 | 33,446 1.3874 | 1.3929 36.389 | 1.3984 39.266 | 1.4039 42.080 |
| 1.3820 30.489 | 33.500 1.3875 | 1.3930 36.442 | 1.3985 39.318 | 1.4040 42.130 |
| 1.3821 30.544 | 33.555 1.3876 | 1.3931 36.495 | 1.3986 39,370 | 1.4041 42.181 |
| 1.3822 30.600 | 33.609 1.3877 | 1.3932 36.548 | 1.3987 39.421 | 1.4042 42.231 |
| 1.3823 30.655 | 33.663 1.3878 | 1.3933 36,601 | 1.3988 39.473 | 1.4043 42.282 |
| 1.3824 30.711 | 33.717 1.3879 | 1.3934 36.654 | 1.3989 39.525 | 1.4044 42.332 |
| 1.3825 30.766 | 33.771 1.3880 | 1.3935 36.706 | 1.3990 39.576 | 1.4045 42.383 |
| 1.3826 30.821 | 33.825 1.3881 | 1.3936 36.759 | 1.3991 39.628 | 1.4046 42.433 |
| 1.3827 30.876 | 33.879 1.3882 | 1.3937 36.812 | 1.3992 39.679 | 1.4047 42.484 |
| 1.3828 30.932 | 33.933 1.3883 | 1.3938 36.865 | 1.3993 39.731 | 1.4048 42.534 |
| 1.3829 30.987 | 33.987 1.3884 | 1.3939 36.917 | 1.3994 39.782 | 1.4049 42.585 |
| 1.3830 31.042 | 34.040 1.3885 | 1.3940 36.970 | 1.3995 39,834 | 1.4050 42.635 |
| 1.3831 31.097 | 34.094 1.3886 | 1.3941 37.023 | 1.3996 39.885 | 1.4051 42.685 |
| 1.3832 31.152 | 34.148 1.3887 | 1.3942 37.075 | 1.3997 39.937 | 1.4052 42.736 |
| 1.3833 31.207 | 34.202 1.3888 | 1.3943 37.128 | 1.3998 39.988 | 1.4053 42.786 |
| 1.3834 31.262 | 34.256 1.3889 | 1.3944 37.180 | 1.3999 40.040 | 1.4054 42.836 |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose |
| Wed C) (%) | st. C) (%) | st. C) ( %) | Wed C) (%) | Wed C) (%) |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| 1.4055 42.887 | 45.623 1.4110 | 1.4165 48.302 | 1.4220 50.928 | 1.4275 53.501 |
| 1.4056 42.937 | 45.672 1.4111 | 1,4166 48350 | 1.4221 50.951 | 1.4276 53.548 |
| 1.4057 42.987 | 45.721 1.4112 | 1.4167 48.399 | 1.4222 51.022 | 1.4277 53.594 |
| 1.4058 43.037 | 45.770 1.4113 | 1.4168 48.447 | 1.4223 51.069 | 1.4278 53.640 |
| 1.1059 43.088 | 45.820 1.4114 | 1.4169 48.495 | 1.4224 51.116 | 1.4279 53.686 |
| 1.4060 43.138 | 45.869 1.4115 | 1.4170 48.543 | 1.4225 51.164 | 1.4280 53.733 |
| 1.4061 43.188 | 45.918 1.4116 | 1.4171 48.591 | 1.4226 51.211 | 1.4281 53.779 |

| 1.4062 43.238 | 46.967 1.4117 | 1.4172 48.639 | 1.4227 51.258 | 1.4282 53.825 |
| 1.4063 43.288 | 46.016 1.4118 | 1.4173 48.687 | 1,4228 51305 | 1.4283 53.871 |
| 1.4064 43.338 | 46.065 1.4119 | 1.4174 48.735 | 1.4229 51.352 | 1.4284 53.918 |
| 1.4065 43.388 | 46.114 1.4120 | 1.4175 48.784 | 1.4230 51.399 | 1.4285 53.964 |
| 1.4066 43.439 | 46.163 1.4121 | 1.4176 48.832 | 1.4231 51.446 | 1.4286 54.010 |
| 1.4067 43.489 | 46.212 1.4122 | 1.4177 48.880 | 1.4232 51.493 | 1.4287 54.056 |
| 1.4068 43.539 | 46.261 1.4123 | 1.4178 48.928 | 1.4233 51.540 | 1.4288 54.102 |
| 1.4069 43.589 | 46.310 1.4124 | 1.4179 48.976 | 1.4234 51.587 | 1.4289 54.148 |
| 1.4070 43.639 | 46.359 1.4125 | 1.4180 49.023 | 1,4235 51,634 | 1.4290 54.194 |
| 1.4071 43.689 | 46.408 1.4126 | 1.4181 49.071 | 1.4236 51.681 | 1.4291 54.241 |
| 1.4072 43.739 | 46.457 1.4127 | 1.4182 49.119 | 1.4237 51.728 | 1.4292 54.287 |
| 1.4073 43.789 | 46.506 1.4128 | 1.4183 49,167 | 1.4238 51.775 | 1.4293 54.333 |
| 1.4074 43.838 | 46.555 1.4229 | 1.4184 49.215 | 1.4239 51.822 | 1.4294 54.379 |
| 1.4075 43.888 | 46.604 1.4130 | 1.4185 49.263 | 1.4240 51.869 | 1.4295 54.425 |
| 1.4076 43.938 | 46.652 1.4131 | 1.4186 49.311 | 1.4241 51.916 | 1.4296 54.471 |
| 1.4077 43.988 | 46.701 1.4132 | 1.4187 49.359 | 1.4242 51.963 | 1.4297 54.517 |
| 1.4078 44.038 | 46.750 1.4133 | 1.4188 49.407 | 1.4243 52.010 | 1.4298 54.563 |
| 1.4079 44.088 | 46.799 1.4134 | 1.4189 49.454 | 1.4244 52.057 | 1.4299 54.609 |
| 1.4080 44.138 | 46.848 1.4135 | 1.4190 49.502 | 1.4245 52.104 | 1.4300 54.655 |
| 1.4081 44.187 | 46.896 1.4136 | 1.4191 49.550 | 1.4246 52.150 | 1.4301 54.701 |
| 1.4082 44.237 | 46.945 1.4137 | 1.4192 49.598 | 1.4247 52.197 | 1.4302 54.746 |
| 1.4083 44.287 | 46.994 1.4138 | 1.4193 49.645 | 1.4248 52.244 | 1.4303 54.792 |
| 1.4084 44.337 | 47.043 1.4139 | 1.4194 49.693 | 1.4249 52.291 | 1.4304 54.838 |
| 1.4085 44.386 | 47.091 1.4140 | 1.4195 49.741 | 1.4250 52.338 | 1.4305 54.884 |
| 1.4086 44.436 | 47.140 1.4141 | 1.4196 49.788 | 1.4251 52.384 | 1.4306 54.930 |
| 1.4087 44.486 | 47.188 1.4142 | 1.4197 49.836 | 1.4252 52.431 | 1.4307 54.976 |
| 1.4088 44.535 | 47.237 1.4143 | 1.4198 49.884 | 1.4253 52.478 | 1.4308 55.022 |
| 1.4089 44.585 | 47.286 1.4144 | 1.4199 49.931 | 1.4254 52.524 | 1.4309 55,067 |
| 1.4090 44.635 | 47.334 1.4145 | 1.4200 49.979 | 1.4255 52.571 | 1.4310 55.113 |
| 1.4091 44.684 | 47.383 1.4146 | 1.4201 50.027 | 1.4256 52.618 | 1.4311 55.159 |
| 1.4092 44.734 | 47.431 1.4147 | 1.4202 50.074 | 1.4257 52.664 | 1.4312 55.205 |
| 1.4093 44.783 | 47.480 1.4148 | 1.4203 50.122 | 1.4258 52.711 | 1.4313 55.250 |
| 1.4094 44.833 | 47.528 1.4149 | 1.4204 50.169 | 1.4259 52.758 | 1.4314 55.296 |
| 1.4095 44.882 | 47.577 1.4150 | 1.4205 50.217 | 1.4260 52.804 | 1.4315 55.342 |
| 1.4096 44.932 | 47.625 1.4151 | 1.4206 50.264 | 1.4261 52.851 | 1.4316 55.388 |
| 1.4097 44.981 | 47.674 1.4152 | 1.4207 50.312 | 1.4262 52.897 | 1.4317 55.433 |
| 1.4098 45.031 | 47.722 1.4153 | 1.4208 50.359 | 1.4263 52.944 | 1.4318 55.479 |
| 1.4099 45.080 | 47.771 1.4154 | 1.4209 50.407 | 1.4264 52.990 | 1.4319 55.524 |
| 1.4100 45.130 | 47.819 1.4155 | 1.4210 50.454 | 1.4265 53.037 | 1.4320 55.570 |
| 45179 1.4101 | 1.4156 47,868 | 1.4211 50.502 | 1.4266 53.083 | 1.4321 55.616 |
| 1.4102 45.228 | 47.916 1.4157 | 1.4212 50.549 | 1.4267 53.130 | 1.4322 55.661 |
| 1.4103 45.278 | 47.964 1.4158 | 1.4213 50.596 | 1.4268 53.176 | 1.4323 55.707 |
| 1.4104 45.327 | 48.013 1.4159 | 1.4114 50.644 | 1.4269 53.223 | 1.4324 55.752 |
| 1.4105 45.376 | 48.061 1.4160 | 1.4215 50.691 | 1.4270 53.269 | 1.4325 55.798 |
| 1.4106 45.426 | 48.109 1.4161 | 1.4216 50.738 | 1.4271 53.316 | 1.4326 55.844 |
| 45475 1.4107 | 1.4162 48,158 | 1.4217 50.786 | 1.4272 53.362 | 1.4327 55.889 |
| 1.4108 45.524 | 48.206 1.4163 | 1.4218 50.833 | 1.4273 53.408 | 1.4328 55.935 |
| 1.4109 45.574 | 48.254 1.4164 | 1.4219 50.880 | 1.4274 53.455 | 1.4329 55.980 |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose |
| Wed C) (%) | st. C) (%) | st. C) ( %) | Wed C) (%) | Wed C) (%) |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| 1.4330 56.026 | 58.503 1.4385 | 1.4440 60.935 | 1.4495 63.324 | 1.4550 65.672 |

| 1.4331 56.071 | 58.547 1.4386 | 1.4441 60.979 | 1.4496 63.367 | 1.4551 65.714 |
| 1.4332 56.116 | 58.592 1.4387 | 1.4442 61.023 | 1.4497 63.410 | 1.4552 65.756 |
| 1.4333 56.162 | 58.637 1.4388 | 1.4443 61.066 | 1.4498 63.453 | 1.4553 65.798 |
| 1.4334 56.207 | 58.681 1.4389 | 1.4444 61.110 | 1.4499 63.496 | 1.4554 65.841 |
| 1.4335 56.253 | 58.726 1.4390 | 1.4445 61.154 | 1.4500 63.539 | 1.4555 65.883 |
| 1.4336 56.298 | 58.770 1.4391 | 1.4446 61.198 | 1.4501 63.582 | 1.4556 65.925 |
| 1.4337 56.343 | 58.815 1.4392 | 1.4447 61.241 | 1.4502 63.625 | 1.4557 65.967 |
| 1.4338 56.389 | 58.859 1.4393 | 1.4448 61.285 | 1.4503 63.668 | 1.4558 66.010 |
| 1.4339 56,434 | 58,904 1.4394 | 1.4449 61.329 | 1.4504 63.711 | 1.4559 66.052 |
| 1.4340 56.479 | 58.948 1.4395 | 1.4450 61.372 | 1.4505 63.754 | 1.4560 66.094 |
| 1.4341 56.525 | 58.993 1.4396 | 1.4451 61.416 | 1.4506 63.797 | 1.4561 66.136 |
| 1.4342 56.570 | 59.037 1.4397 | 1.4452 61.460 | 1.4507 63.840 | 1.4562 66.178 |
| 1.4343 56.615 | 59.082 1.4398 | 1.4453 61.503 | 1.4508 63.882 | 1.4563 66.221 |
| 1.4344 56.660 | 59.126 1.4399 | 1.4454 61.547 | 1.4509 63.925 | 1.4564 66.263 |
| 1.4345 56.706 | 59.170 1.4400 | 1.4455 61.591 | 1.4510 63.968 | 1.4565 66.305 |
| 1.4346 56.751 | 59.215 1.4401 | 1.4456 61.634 | 1.4511 64.011 | 1.4566 66.347 |
| 1.4347 56.796 | 59.259 1.4402 | 1.4457 61.678 | 1.4512 64.054 | 1.4567 66.389 |
| 1.4348 56.841 | 59.304 1.4403 | 1.4458 61.721 | 1.4513 64.097 | 1.4568 66.431 |
| 1.4349 56.887 | 59.348 1.4404 | 1.4459 61.765 | 1.4514 64.139 | 1.4569 66.473 |
| 1.4350 56.932 | 59.392 1.4405 | 1.4460 61.809 | 1.4515 64.182 | 1.4570 66.515 |
| 1.4351 56.977 | 59.437 1.4406 | 1.4461 61.852 | 1.4516 64.225 | 1.4571 66.557 |
| 1.4352 57.022 | 59.481 1.4407 | 1.4462 61.896 | 1.4517 64.268 | 1.4572 66.599 |
| 1.4353 57.067 | 59.525 1.4408 | 1.4463 61.939 | 1.4518 64.311 | 1.4573 66.641 |
| 1.4354 57.112 | 59.569 1.4409 | 1.4464 61.983 | 1.4519 64.353 | 1.4574 66.683 |
| 1.4355 57.157 | 59.614 1.4410 | 1.4465 62.026 | 1.4520 64.396 | 1.4575 66.725 |
| 1.4356 57.202 | 59.658 1.4411 | 1.4466 62.070 | 1.4521 64.439 | 1.4576 66.767 |
| 1.4357 57.247 | 59.702 1.4412 | 1.4467 62.113 | 1.4522 64.481 | 1.4577 66.809 |
| 1.4358 57.292 | 59.746 1.4413 | 1.4468 62.156 | 1.4523 64.524 | 1.4578 66.851 |
| 1.4359 57.337 | 59.791 1.4414 | 1.4469 62.200 | 1.4524 64.567 | 1.4579 66.893 |
| 1.4360 57.382 | 59.835 1.4415 | 1.4470 62.243 | 1.4525 64.609 | 1.4580 66.935 |
| 1.4361 57.427 | 59.879 1.4416 | 1.4471 62.287 | 1.4526 64.652 | 1.4581 66.977 |
| 1.4362 57.472 | 59.923 1.4417 | 1.4472 62.330 | 1,45Z7 64695 | 1.4582 67.019 |
| 1.4363 57.517 | 59.967 1.4418 | 1.4473 62.373 | 1.4528 64.737 | 1.4583 67.061 |
| 1.4364 57.562 | 60.011 1.4419 | 1.4474 62.417 | 1.4529 64.780 | 1.4584 67.103 |
| 1.4365 57.607 | 60.056 1.4420 | 1.4475 62.460 | 1.4530 64.823 | 1.4585 67.145 |
| 1.4366 57.652 | 60.100 1.4421 | 1.4476 62.503 | 1.4531 64.865 | 1.4586 67.186 |
| 1.4367 57.697 | 60.144 1.4422 | 1.4477 62.547 | 1.4532 64.908 | 1.4587 67.228 |
| 1.4368 57.742 | 60.188 1.4423 | 1.4478 62.590 | 1.4533 64.950 | 1.4588 67.270 |
| 1.4369 57.787 | 60.232 1.4424 | 1.4479 62.633 | 1.4534 64.993 | 1.4589 67.312 |
| 1.4370 57.832 | 60.276 1.4425 | 1.4480 62.677 | 1.4535 65.035 | 1.4590 67.354 |
| 1.4371 57.877 | 60.320 1.4426 | 1.4481 62.720 | 1.4536 65.078 | 1.4591 67.396 |
| 1.4372 57.921 | 60.364 1.4427 | 1.4482 62.763 | 1.4537 65.120 | 1.4592 67.437 |
| 1.4373 57.966 | 60.408 1.4428 | 1.4483 62.806 | 1.4538 65.163 | 1.4593 67.479 |
| 1.4374 58.011 | 60.452 1.4429 | 1.4484 62.849 | 1.4539 65.205 | 1.4594 67.521 |
| 1.4375 58.056 | 60.496 1.4430 | 1.4485 62.893 | 1.4540 65.248 | 1.4595 67.563 |
| 1.4376 58.101 | 60.540 1.4431 | 1.4486 62.936 | 1.4541 65.290 | 1.4596 67.604 |
| 1.4377 58.145 | 60.584 1.4432 | 1.4487 62.979 | 1.4542 65.333 | 1.4597 67.640 |
| 1.4378 58.190 | 60.628 1.4433 | 1.4488 63.022 | 1.4543 65.375 | 1.4598 67.688 |
| 1.4379 58.235 | 60.672 1.4434 | 1.4489 63.065 | 1.4544 65.417 | 1.4599 67.729 |
| 1.4380 58.279 | 60.716 1.4435 | 1.4490 63.108 | 1.4545 65.460 | 1.4600 67.771 |
| 1.4381 58.324 | 60.759 1.4436 | 1.4491 63.152 | 1.4546 65.502 | 1.4601 67.813 |
| 1.4382 58.369 | 60.803 1.4437 | 1.4492 63.195 | 1.4547 65.544 | 1.4602 67.854 |
| 1.4383 58.413 | 60.847 1.4438 | 1.4493 63.238 | 1.4548 65.587 | 1.4603 67.896 |

| 1.4384 58.458 | 60.891 1.4439 | 1.4494 63.281 | 1.4549 65.629 | 1.4604 67.938 |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose |
| Wed C) (%) | st. C) (%) | st. C) ( %) | Wed C) (%) | Wed C) (%) |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| 1.4605 67.979 | 70.249 1.4660 | 1.4715 72.482 | 1.4770 74.678 | 1.4825 76.841 |
| 1.4606 68.021 | 70.290 1.4661 | 1.4716 72.522 | 1.4771 74.718 | 1.4826 76.880 |
| 1.4607 68.063 | 70.331 1.4662 | 1.4717 72.562 | 1.4772 74.758 | 1.4827 76.919 |
| 1.4608 68.104 | 70.372 1.4663 | 1.4718 72.602 | 1.4773 74.797 | 1.4828 76.958 |
| 1.4609 68.146 | 70.413 1.4664 | 1.4719 72.643 | 1.4774 74.837 | 1.4829 76.997 |
| 1.4610 68 187 | 70 453 1,4665 | 1.4720 72.683 | 1.4775 74.876 | 1.4830 77.036 |
| 1.4611 68.229 | 70.494 1.4666 | 1.4721 72.723 | 1.4776 74.916 | 1.4831 77.075 |
| 1.4612 68.270 | 70.535 1.4667 | 1.4722 72.763 | 1.4777 74.956 | 1.4832 77.113 |
| 1.4613 68.312 | 70.576 1.4668 | 1.4723 72.031 | 1.4778 74.995 | 1.4833 77.152 |
| 1.4614 68.353 | 70.617 1.4669 | 1.4724 72.843 | 1.4779 75.035 | 1.4834 77.191 |
| 1.4615 68.395 | 70.658 1.4670 | 1.4725 72.884 | 1.4780 75.074 | 1.4835 77.230 |
| 1.4616 68.436 | 70.698 1.4671 | 1.4726 72.924 | 1.4781 75.114 | 1.4836 77.269 |
| 1.4617 68.478 | 70.739 1.4672 | 1.4727 72.964 | 1.4782 75.153 | 1.1837 77.308 |
| 1.4618 68.519 | 70.780 1.4673 | 1.4728 73.004 | 1.4783 75.193 | 1.4838 77.347 |
| 1.4619 68.561 | 70.821 1.4674 | 1.4729 73.044 | 1.4784 75.232 | 1.4839 77.386 |
| 1.4620 68.602 | 70.861 1.4675 | 1.4730 73.084 | 1.4785 75.272 | 1.4840 77.425 |
| 1.4621 68.643 | 70.902 1.4676 | 1.4731 73.124 | 1.4786 75.311 | 1.4841 77.463 |
| 1.4622 68.685 | 70.943 1.4677 | 1.4732 73.164 | 1.4787 75.350 | 1.4842 77.502 |
| 1.4623 68.726 | 70.984 1.4678 | 1.4733 73.204 | 1.4788 75.390 | 1.4843 77.541 |
| 1.4624 68.768 | 71.024 1.4679 | 1.4734 73.244 | 1.4789 75.429 | 1.3844 77.580 |
| 1.4625 68.809 | 71.065 1.4680 | 1.4735 73.285 | 1.4790 75.469 | 1.4845 77.619 |
| 1.4626 68.850 | 71.106 1.4681 | 1.4736 73.325 | 1.4791 75.508 | 1.4846 77.657 |
| 1.4627 68.892 | 71.146 1.4682 | 1.4737 73.365 | 1.4792 75.547 | 1.4847 77.696 |
| 1.4628 68.933 | 71.187 1.4683 | 1.4738 73.405 | 1.4793 75.587 | 1.4848 77.735 |
| 1.4629 68.974 | 71.228 1.4684 | 1.4739 73.445 | 1.4794 75.626 | 1.4849 77.774 |
| 1,4630 69016 | 71268 1.4685 | 1.4740 73.485 | 1.4795 75.666 | 1.4850 77.812 |
| 1.4631 69.057 | 71.309 1.4686 | 1.4741 73.524 | 1.4796 75.705 | 1.4851 77.851 |
| 1.4631 69.098 | 71.349 1.4687 | 1.4742 73.564 | 1.4797 75.744 | 1.4852 77.890 |
| 1.4633 69.139 | 71.390 1.4688 | 1.4743 73.604 | 1.4798 75.784 | 1.4853 77.928 |
| 1.4634 69.181 | 71.431 1.4689 | 1.4744 73.644 | 1.4799 75.823 | 1.4854 77.967 |
| 1.4635 69.222 | 71.471 1.4690 | 1.4745 73.684 | 1.4800 75.862 | 1.4855 78.006 |
| 1.4636 69.263 | 71.512 1.4691 | 1.4746 73.724 | 1.4801 75.901 | 1.4856 78.045 |
| 1.4637 69.304 | 71.552 1.4692 | 1.4747 73.764 | 1.4802 75.941 | 1.4857 78.083 |
| 1.4638 69.346 | 71.593 1.4693 | 1.1748 73.804 | 1.4803 75.980 | 1.4858 78.122 |
| 1.4639 69.387 | 71.633 1.4694 | 1.4749 73.844 | 1.4804 76.019 | 1.4859 78.160 |
| 1.4640 69.428 | 71.674 1.4695 | 1.4750 73.884 | 1.4805 76.058 | 1.4860 78.199 |
| 1.4641 69.469 | 71.714 1.4696 | 1.4751 73.924 | 1.4806 76.098 | 1.4861 78.238 |
| 1.4642 69.510 | 71.755 1.4697 | 1.4752 73.963 | 1.4807 76.137 | 1.4862 78.276 |
| 1.4643 69.551 | 71.795 1.4698 | 1.4753 74.003 | 1.4808 76.176 | 1.4863 78.315 |
| 1.4644 69.593 | 71.836 1.4699 | 1.4754 74.043 | 1.4809 76.215 | 1.4864 78.353 |
| 1.4645 69.634 | 71.876 1.4700 | 1.4755 74.083 | 1.4810 76.254 | 1.4865 78.392 |
| 1.4646 69.675 | 71.917 1.4701 | 1.4756 74.123 | 1.4811 76.294 | 1.4866 78.431 |
| 1.4647 69.716 | 71.957 1.4702 | 1.4757 74.162 | 1.4812 76.333 | 1.4867 78.469 |
| 1.4648 69.757 | 71.998 1.4703 | 1.4758 74.202 | 1.4813 76.372 | 1.4868 78.508 |
| 1.4649 69.798 | 72.038 1.4704 | 1.4759 74.242 | 1.4814 76.411 | 1.4869 78.546 |
| 1.4650 69.839 | 72.078 1.4705 | 1.4760 74.282 | 1.4815 76.450 | 1.4870 78.585 |
| 1.4651 69.880 | 72.119 1.4706 | 1.4761 74.321 | 1.4816 76.489 | 1.4871 78.623 |
| 1.4652 69.921 | 72.159 1.4707 | 1.4762 74.361 | 1.4817 76.528 | 1.4872 78.662 |

| 1.4653 69.962 | 72.199 1.4708 | 1.4763 74.401 | 1.4818 76.567 | 1.4873 78.700 |
| 1.4654 70.003 | 72.240 1.4709 | 1.4764 74.441 | 1.4819 76.607 | 1.4874 78.739 |
| 1.4655 70.044 | 72.280 1.4710 | 1.4765 74.480 | 1.4820 76.646 | 1.4875 78.777 |
| 1.4656 70.085 | 72.320 1.4711 | 1.4766 74.520 | 1.4821 76.685 | 1.4876 78.816 |
| 1.4657 70.126 | 72.361 1.4712 | 1.4767 74.560 | 1.4822 76.724 | 1.4877 78.854 |
| 1.4658 70.167 | 72.401 1.4713 | 1.4768 74.599 | 1.4823 76.763 | 1.4878 78.892 |
| 1.4659 70.208 | 72.441 1.4714 | 1.4769 74.639 | 1.4824 76.802 | 1.4879 78.931 |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose | n (20 Sucrose |
| Wed C) (%) | st. C) (%) | st. C) ( %) | Wed C) (%) | Wed C) (%) |
+-------------------+-----------------+-------------------+-------------------+--------------------+
| 1.4880 78.969 | 80.497 1.4920 | 1.4960 82.007 | 1.5000 83.500 | 1.5040 84.976 |
| 1.4881 79.008 | 80.534 1.4921 | 1.4961 82.044 | 1.5001 83.537 | 1.5041 85.013 |
| 1.4882 79.046 | 80.572 1.4922 | 1.4962 82.082 | 1.5002 83.574 | 1.5042 85.049 |
| 1.4883 79.084 | 80.610 1.4923 | 1.4963 82.119 | 1.5003 83.611 | 1.5043 85.086 |
| 1.4884 79.123 | 80.648 1.4924 | 1.4964 82.157 | 1.5004 83.648 | 1.5044 85.123 |
| 1.4885 79.161 | 80.686 1.4925 | 1.4965 82.194 | 1.5005 83.685 | 1.5045 85.159 |
| 1.4886 79.199 | 80.724 1.4926 | 1.4966 82.232 | 1.5006 83.722 | 1.5046 85.196 |
| 1.4887 79.238 | 80.762 1.4927 | 1.4967 82.269 | 1.5007 83.759 | 1.5047 85.233 |
| 1.4888 79.276 | 80.800 1.4928 | 1.4968 82.307 | 1.5008 83.796 | 1.5048 85.269 |
| 1.4889 79.314 | 80.838 1.4929 | 1.4969 82.344 | 1.5009 83.833 | 1.5049 85.306 |
| 1.4890 79.353 | 80.876 1.4930 | 1.4970 82.381 | 1.5010 83.870 | 1.5050 85.343 |
| 1.4891 79.391 | 80.913 1.4931 | 1.4971 82.419 | 1.5011 83.907 | 1.5051 85.379 |
| 1.4892 79.429 | 80.951 1.4932 | 1.4972 82.456 | 1.5012 83.944 | 1.5052 85.416 |
| 1.4893 79.468 | 80.989 1.4933 | 1.4973 82.494 | 1.5013 83.981 | 1.5053 85.452 |
| 1.4894 79.506 | 81.027 1.4934 | 1.4974 82.531 | 1.5014 84.018 | 1.5054 85.489 |
| 1.4895 79.544 | 81.065 1.4935 | 1.4975 82.569 | 1.5015 84.055 | 1.5055 85.525 |
| 1.4596 79.582 | 81.103 1.4936 | 1.4976 82.606 | 1.5016 84.092 | 1.5056 85.562 |
| 1.4597 79.620 | 81.140 1.4937 | 1.4977 82.643 | 1.5017 84.129 | 1.5057 85.598 |
| 1.4898 79.659 | 81.178 1.4938 | 1.4978 32.681 | 1.5018 84.166 | 1.5058 85.635 |
| 1.4899 79.697 | 81.216 1.4939 | 1.4979 82.718 | 1.5019 84.203 | 1.5059 85.672 |
| 1.4900 79.735 | 81.254 1.4940 | 1.4980 82.755 | 1.5020 84.240 | 1.5060 85.708 |
| 1.4901 79.773 | 81.291 1.4941 | 1.4981 82.793 | 1.5021 84.277 | 1.5061 85.744 |
| 1.4902 79.811 | 81.329 1.4942 | 1.4982 82.830 | 1.5022 84.314 | 1.5062 85.781 |
| 1.4903 79.850 | 81.367 1.4943 | 1.4983 82.867 | 1.5023 84.351 | 1.5063 85.817 |
| 1.4904 79.888 | 81.405 1.4944 | 1.4984 82.905 | 1.5024 84.388 | 1.5064 85.854 |
| 1.4905 79.926 | 81.442 1.4945 | 1.4985 82.942 | 1.5025 84.424 | 1.5065 85.890 |
| 1.4906 79.964 | 81.480 1.4946 | 1.4986 82.979 | 1.5026 84.461 | 1.5066 85.927 |
| 1.4907 80.002 | 81.518 1.4947 | 1.4987 83.016 | 1.5027 84.498 | 1.5067 85.963 |
| 1.4908 80.040 | 81.555 1.4948 | 1.4988 83.054 | 1.5028 84.535 | 1.5068 86.000 |
| 1.4909 80.078 | 81.593 1.4949 | 1.4989 83.091 | 1.5029 84.572 | 1.5069 86.036 |
| 1.4910 80.116 | 81.631 1.4950 | 1.4990 83.128 | 1.5030 84.609 | 1.5070 86.072 |
| 1.4911 80.154 | 81.668 1.4951 | 1.4991 83.165 | 1.5031 84.645 | 1.5071 86.109 |
| 1.4912 80.192 | 81.706 1.4952 | 1.4992 83.202 | 1.5032 84.682 | 1.5072 86.145 |
| 1.4913 80.231 | 81.744 1.4953 | 1.4993 83.240 | 1.5033 84.719 | 1.5073 86.182 |
| 1.4914 80.269 | 81.781 1.4954 | 1.4994 83.277 | 1.5034 84.756 | 1.5074 86.218 |
| 1.4915 80.307 | 81.819 1.4955 | 1.4995 83.314 | 1.5035 84.792 | 1.5075 86.254 |
| 1.4916 80.345 | 81.856 1.4956 | 1.4996 83.351 | 1.5036 84.829 | 1.5076 86.291 |
| 1.4917 80.383 | 81.894 1.4957 | 1.4997 83.388 | 1.5037 84.866 | 1.5077 86.327 |
| 1.4918 80.421 | 81.932 1.4958 | 1.4998 83.425 | 1.5038 84.903 | 1.5078 86.363 |
| 1.4919 80.459 | 81.969 1.4959 | 1.4999 83.463 | 1.5039 84.939 | 1.5079 86.399 |
+-------------------+-----------------+-------------------+-------------------+--------------------+

1) The values ​​of n in these tables are calculated by equations, which

Author for the ICUMSA K. Rosenhauer. The formula was programmed and performed calculations
Frank G. Carpenter from USDA and published in Sugar
J. 33, 15-22 (June 1970). The refractive index was measured at 20 Wed.
C using the sodium D line Brix (% ​​by weight of sucrose) was obtained
weighing at 20 Wed. C in air at a pressure of 760 torr (mm Hg) and 50% relative humidity
. This table replaces the previous table (47012, 11
edition) published in Int. Sugar J. 39, 22S (1937).


Příl.40
Methods of sampling for the official control of tin in foods packaged in cans


First Purpose and scope

Samples intended for official control of the levels of tin in foods packaged in cans
be taken of the following methods.
Thus obtained aggregate samples shall be considered as representative of that batch.
Compliance with maximum limits laid down in Commission Regulation (EC) no. 466/2001
determined on the basis of the levels determined in the laboratory samples.

Second definition

Lot: an identifiable quantity of food delivered at one time
which, according to the person referred to in § 3 para. 1
uniform characteristics such as origin, variety, type of packing, packer, consignor or
designation.

Part of the batch: batch certain part allocated to it was carried
sampling. Each sublot must be physically separate and identifiable
.

Incremental sample: a quantity of material taken from a single place in the lot or sublot
.

Aggregate sample: the combined total of all the incremental samples taken from the lot or sublot
.

Laboratory sample: sample intended for the laboratory.

Third General provisions
3.1
Staff

Sampling must be done by a person meeting the requirements specified in § 3
paragraph. 1st

3.2 Material to be removed

Each lot which is to be examined must be sampled separately.

3.3 Precautions

Of sampling and preparation of the samples must be pre
measures to avoid any changes, which could affect
tin content, adversely affect the analytical determination or degrade
representativeness aggregate samples.
Sub-samples 3.4


Incremental samples shall, if possible, from various places throughout the lot or sublot
. Deviations from this procedure must be recorded in the log.

3.5 Preparation of aggregate sample

Aggregate sample is made up by uniting all incremental samples and their
homogenization in the laboratory.

3.6 Replicate laboratory samples

Replicate samples for testing to confirm, defense
business and referee purposes shall be taken from the homogenised aggregate sample
.

3.7 Packaging and transmission of samples

Each sample is placed in a clean, inert container which provides
protection from contamination and against damage in transit.
Must be taken all the necessary precautions to avoid any change
composition of the sample, which might arise during transportation or storage.

3.8 Sealing and labeling of samples

Each sample taken for official use shall be sealed at the place of sampling and identified following
§ 6 of each sampling record must be kept according to § 5.


Fourth Sampling plans

The sampling method applied shall ensure that the aggregate sample is representative for
controlled batch.

4.1 Number of subsamples

Minimum number of incremental samples to be taken from cans
lot is given in the table. Incremental samples taken from each can
must be of similar weight and has an aggregate sampling. Table
:

Number of cans (incremental samples) which shall be taken,
to form the aggregate sample

-------------------------------------------------- --------
The number of cans in the lot Number of cans, which must
or part of the batch to be removed
-------------------------------------------------- --------
1 to 25 of at least one tin
-------------------------------------------------- --------
26-100 least two cans
-------------------------------------------------- --------
> 100 5 cans
-------------------------------------------------- --------

Maximum levels apply to the contents of each can. For practical

Reasons for the examinations necessary to create a bulk sample. If
result for the aggregate sample exceeds the maximum limit, but close
to the maximum limit, and if it is suspected that individual cans may be
maximum limit is exceeded, can be done
further examination.

4.2 Sampling at retail

Sampling at retail stage shall be done in accordance with the above provisions
sampling or other procedures according to § 1 para. 2, § 3 and §
4. These procedures must be adequately sampled lot
representative.

Fifth Compliance with the specifications of the lot or sublot

For confirmation purposes performs control laboratory
least two independent tests and the results averaged. The lot is accepted does not go
diameter of the respective maximum limit specified in Regulation (EC) no. 466/2001
taking into account the measurement uncertainty and correction for recovery.

The lot is rejected if taking into account the measurement uncertainty and correction for recovery
diameter exceeds the maximum limit specified in Regulation
(EC) no. 466/2001.


Příl.41
Methods of sampling for the official control of the levels of patulin in certain foodstuffs


First Purpose and scope

Samples for the official control of the levels of patulin in foodstuffs shall be taken
methods described below. Aggregate samples thus obtained are
considered as representative of the lots.
Compliance with maximum limits laid down in Commission Regulation (EC) no. 466/2001 shall be based on
levels determined in the laboratory samples.

Second definition

Lot: an identifiable quantity of food delivered at one time,
which, according to the person referred to in § 3 para. 1 common characteristics,
such as origin, variety, type of packing, packer, consignor or markings.

Part of the batch: batch certain part allocated to it was carried
sampling. Each sublot must be physically separate and identifiable
.

Incremental sample: a quantity of material taken from a single place in the lot or sublot
.

Aggregate sample: the combined total of all the incremental samples taken from the lot or sublot
.

Third General provisions
3.1
Staff

Sampling is done by an authorized person (§ 3 para. 1).

3.2 Material to be removed

Each lot which is to be examined must be sampled separately.

3.3 Precautions

Of sampling and sample preparation must be done
precautions to avoid any changes which would affect the content
patulin, adversely affect the analytical determination or degrade
representativeness aggregate samples.
Sub-samples 3.4


Individual samples were taken from various places throughout the lot or sublot.
Any deviation from this procedure must be recorded in the log.

3.5 Preparation of aggregate sample

Aggregate sample of at least 1 kg is prepared by uniting the incremental samples
unless carried out sampling of individual packages weighing more than 1 kg
.
3.6 Replicate samples


Replicate samples for enforcement, defense
commercial dispute or arbitration shall be taken from the homogenised aggregate sample
.

3.7 Packaging and transmission of samples

Each sample is placed in a clean, inert container which
provides adequate protection from contamination and damage during transportation
. They have taken all necessary measures to prevent
change in composition of the sample, which might arise during transportation or storage.

3.8 Sealing and labeling of samples

Each sample taken for official use shall be sealed at the place of sampling and identified following
§ 6 of each sampling record must be kept
Sampling according to § 5

Fourth Sampling plans

The sampling method applied ensures that the aggregate sample is representative for
controlled batch.
Number of subsamples


The aggregate sample is prepared in accordance with paragraph 3.5.

The minimum number of incremental samples to be taken from the lot is
shown in Table 1. For liquid products before taking samples
as thoroughly manually or mechanically mixed, assumes a uniform extension
patulin in a given lot. To create summary

Sample for liquid products because enough of each batch to take three incremental samples
.

Incremental samples should be of similar weight. Weight of the incremental sample should be at least 100 grams
that uniting the incremental samples originated aggregate sample
weighing more than 1 kg. Deviation from this procedure must be recorded
in the sampling protocol.
Table 1:
Minimum number of incremental samples to be taken from
batch

-------------------------------------------------- --------
Weight of lot Minimum number of incremental samples
(Kg) to be removed
-------------------------------------------------- --------
<3 50
50-500 5
> 500 10
-------------------------------------------------- --------

If the lot consists of individual packages, the number of packages subscribed for
to form the aggregate sample given in Table 2. Table 2
:
Number of packages (incremental samples) collected for the purpose
form the aggregate sample if the lot consists of
individual packages

-------------------------------------------------- --------
Number of packages or units Number of packages or units
in the lot to be sampled
-------------------------------------------------- --------
1 to 25 1 package or unit
26-100 about 5%, at least 2 packages
or units
> 100 about 5%, up to 10 packages
or units
-------------------------------------------------- --------

Fifth Compliance with the specifications of the lot or sublot

Control laboratory performs a re-test laboratory sample for confirmation purposes
result if the result of that received during
first test, 20% lower or higher than the maximum limit, and calculates
average of the results .

The lot is accepted if the result of the first test is 20% lower than
maximum limit, or if it is necessary retest if it meets the relevant
average maximum limit laid down in Regulation (EC) No. || | 466/2001 taking into account the measurement uncertainty and correction for recovery.
Batch is rejected if the average exceeds taking into account the measurement uncertainty and correction for recovery
maximum limit laid down in Regulation (EC) no. 466/2001
.


Příl.42
Sample preparation and requirements for methods of analysis used for official control
tin content in foods packaged in cans

First Precautions and general considerations for tin

Basic requirement is to obtain a representative and homogeneous laboratory
sample without introducing secondary contamination.

Analyst shall ensure that in preparing the samples do not become contaminated
. Apparatus coming into contact with the sample should
be made of inert materials, for example plastics such as polypropylene or polytetrafluoroethylene
and should be cleaned after use
acid to minimize the risk of contamination.
Cutting tools they must be made from high quality stainless steel.

All the collected amount of food received by the laboratory is used to prepare
sample. Reproducible results provide only
thoroughly homogenized samples. You can also use other methods for preparing samples
according to § 1.

Second Treatment of the sample received in the laboratory

Complete aggregate sample is ground finely and mixed thoroughly using a process
which achieves complete homogenisation.

Third Distribution of samples for enforcement and defense purposes

Replicate samples for enforcement, defense
business and referee purposes shall be taken from the homogenised sample.

Fourth Methods of testing and laboratory control requirements
4.1 Definitions


The following are a few common definitions that the laboratory
use.

R = repeatability; value below which will be as expected with the
probability (typically 95%) lie absolute value of the difference between the results of two separate
specific probability (ie.
same sample, same operator, same apparatus, same laboratory,
determined shortly after each other); r = 2.8 x sr,

Sr = standard deviation calculated from results generated under conditions
repeatability

RSDr = relative standard deviation calculated from results

Repeatability conditions [(sr / x) x 100], where x is the average of results
all laboratories and samples

R = reproducibility; value below which will be as expected with the
probability (typically 95%) lie
absolute difference between single test results obtained under reproducibility conditions
(ie. on identical material obtained by operators in different laboratories, as
using the standardized test method); R = 2.8 x sR

SR = standard deviation calculated from results generated under conditions
reproducibility

RSDR = relative standard deviation calculated from results
reproducibility conditions [(SR / x) x 100]

HORRATr = value found RSDr divided by the RSDr value calculated from the Horwitz equation
assuming r = 0.66 R,

HORRATR = the observed RSDR value divided by the RSDR value calculated from
Horwitz equation (2).

U = the expanded uncertainty, using a coverage factor of 2
which gives a level of confidence of approximately 95%.
4.2 General requirements


Testing methods used for food control purposes must comply with § 9.

4.3 Specific requirements


If not directly applicable European Community set
specific methods for the determination of tin in foods packaged in cans
, laboratories may select a validated method provided
selected method meets the criteria listed in the table. When validating
would be the use of certified reference material.
Table: Performance characteristics of analytical methods for
tin

-------------------------------------------------- --------
Parameter Value and comment
-------------------------------------------------- --------
Applicability Foods specified in Regulation
(EC) no. 242/2004
-------------------------------------------------- --------
The lower limit of detection or equal to 5 mg / kg
-------------------------------------------------- --------
The lower limit of quantification or equal to 10 mg / kg
-------------------------------------------------- --------
The accuracy or HORRATr HORRATR achieved
in the validation collaborative study
must be less than 1.5
-------------------------------------------------- --------
The yield of 80% to 105% (reached
in collaborative study)
-------------------------------------------------- --------
Specificity determination must not be interfered
matrix and spectral phenomena
-------------------------------------------------- --------

4.3.1 Operational characteristics - the concept of uncertainty

Suitability test methods to be used in the laboratory, can be assessed
also use the concept of uncertainty.
Laboratory may use a method which will produce results within a maximum standard uncertainty
. The maximum standard uncertainty is calculated using the equation:
Uf = square root of [(LOD / 2) 2 + (0,1C) 2]


Where:
Uf is the maximum standard uncertainty
LOD is the limit of detection methods
C is the concentration.

If a test method provides results with uncertainty measurements less than the maximum
standard uncertainty, the method will be suitable as well as
method which meets the performance characteristics listed in the table.

4.4 Recovery calculation and reporting of results

The test results are reported corrected or uncorrected for recovery.
Must The manner of yield and its value.
Test result corrected for recovery is used for checking compliance.

It should be taken into account harmonized recommendations for the use
recovery in analytical measurement, developed by international
standardization organizations and professional associations, which will help in determining
recovery factors.

The analytical result must be reported as (x +/- U), where x is
analytical result and U is the measurement uncertainty.

4.5 Requirements for laboratories

Laboratories must comply with the special regulation. ^ *)

4.6 Other considerations for testing

Proficiency

It is recommended to participate in appropriate programs proficiency
under international harmonized protocol for the evaluation of professional
levels of chemical analytical laboratories drawn up under the auspices

International standardization organizations and professional associations.

Some of these programs are focused on the determination of tin in foods
and participate in such a program is given priority over participation in general
programs for the determination of metals in foods.
Internal quality control


Laboratories should be able to demonstrate that they have
own internal quality control procedures. Examples in this respect are listed in
recommendations of international standards organizations and professional associations
for internal quality control in analytical chemical laboratories
.
Sample preparation


Must be taken to ensure that all the tin in the sample for the test
converted into solution. Sample dissolution procedure is to ensure that
avoid precipitation hydrolysed compounds of tetravalent tin (dreamy)
(i.e. stannic oxide SnO2, Sn (OH) 4, SnO2.H2O).

The prepared sample was stored in an environment of HCl 5 mol / l.
Due to easy volatility SnCl4, the sample is allowed to boil.


Příl.43
Sample preparation and criteria for test methods used for official
control of the levels of patulin in certain foodstuffs

First Precautionary measures

Patulin may be extended in certain foods unevenly and samples
should therefore be prepared and blended with extreme care.
All the collected amount of food received by the laboratory is used to prepare
sample.

Second Treatment of the sample received in the laboratory

Complete aggregate sample is ground finely and mixed thoroughly using a process
which achieves complete homogenisation.

Third Distribution of samples for testing to confirm the result, and the defense


Replicate samples for enforcement, defense
business and referee purposes shall be taken from the homogenised sample
.

Fourth Test methods used by the laboratory, and management requirements to laboratories

4.1 Definitions


The following are a few common definitions that the laboratory
use.

The most commonly quoted precision parameters are repeatability and reproducibility
.

R = repeatability, the value below which will be as expected with the
probability (typically 95%) lie
absolute difference between single test results under repeatability conditions (ie.
Same sample, same operator , same apparatus, same laboratory,
determined in quick succession) and hence r = 2.8 x sr,

Sr = standard deviation calculated from results generated under conditions
repeatability

RSDr = relative standard deviation calculated from results generated under
repeatability conditions [(sr / x) x 100], where x is the average of results
all laboratories and samples

R = reproducibility; value below which will be as expected with the
probability (typically 95%) lie
absolute difference between single test results obtained under reproducibility conditions
(ie. on identical material obtained by operators in different laboratories using
standardized test method); R = 2.8 x sR

SR = standard deviation calculated from results generated under conditions
reproducibility

RSDR = relative standard deviation calculated from results
reproducibility conditions [(SR / x) x 100]
4.2 General requirements


Testing methods used for food control purposes must comply whenever
possible, in accordance with § 9.
4.3 Specific requirements


If not directly applicable European Community set
specific methods for determining the levels of patulin in foodstuffs may choose
laboratory method provided it meets the following criteria
:
Performance characteristics for patulin | ||
-------------------------------------------------- --------
Levels of patulin
microg / kg -------------------------------------------
RSDr% RSDR% Recovery%
-------------------------------------------------- --------
<= 20 <= 30 <40 50-120
-------------------------------------------------- --------
20-50 = <= 20 <30 70-105
-------------------------------------------------- --------
> = 50 <= 15 <25 75-105
-------------------------------------------------- --------


Detection limits of the methods used are not stated as the precision
given at the concentrations. The precision values ​​
calculated from the Horwitz equation:
(1-0,5logC)
RSDR = 2


Where:

RSDR relative standard deviation calculated from
the results obtained under the conditions
Reproducibility [sR / x) x 100]

C is the concentration ratio (i.e. 1 = 100 g / 100 g, 0.001
= 1000 mg / kg).

This is a generalized equation for the precision with which it was shown that in most routine methods of analysis
independent of analyte and matrix but only
concentration.

4.4 Recovery calculation and reporting of results

As a result of the tests show the result with or without correction for
recovery. Must be The manner of yield and its value.
For checking compliance applies the test result with correction
recovery (see Annex I, section 5).

The test results are given in the form (x +/- U), where x is the result of the test
and U is the measurement uncertainty.

4.5 Requirements for laboratories

Laboratories must comply with the special regulation. ^ *)


Příl.44
Methods of sampling for the official control of benzo [a] pyrene in foodstuffs


First Purpose and scope

Samples for the official control of benzo [a] pyrene in food are collected
methods described below. Aggregate samples thus obtained are
considered as representative of the lots.
Compliance with maximum limits laid down in Commission Regulation (EC) no. 466/2001 shall be based on
levels determined in the laboratory samples.

Second definition

For the purposes of this Annex batches
identifiable quantity of food delivered at one time and determined by the person referred to in paragraph 3 §
. 1 common characteristics, such as origin, variety, type of packaging
packer, consignor or markings.

Third General provisions

3.1 Material to be removed

Each lot which is to be examined must be sampled separately.

3.2 Precautions

Of sampling and sample preparation must be done
precautions to avoid any changes which would affect the content
benzo [a] pyrene, adversely affect the analytical determination or
aggregate samples unrepresentative.
Sub-samples 3.3


Incremental samples shall, if possible, from various places throughout the lot or sublot
. Any deviation from this procedure must be recorded in the log
.

3.4 Preparation of aggregate sample

The aggregate sample is made up by uniting the incremental samples. This sample was homogenized in a laboratory
.

3.5 Replicate laboratory samples

Replicate samples for enforcement, defense
commercial dispute or arbitration shall be taken from the homogenised aggregate sample
.

3.6 Packaging and transport of samples

Each sample is placed in a clean, inert container which
provides adequate protection from contamination and damage during transportation
. They need to be taken all the necessary measures to prevent
change in composition of the sample, which might arise during transportation or storage.

3.7 Sealing and labeling of samples

Each sample taken for official use shall be sealed at the place of sampling and identified following
§ 6

Of each sampling record must be kept for sampling by
§ fifth

Fourth Sampling plans

The sampling method applied ensures that the aggregate sample is representative for
controlled batch.

4.1 Number of subsamples

In the case of oils, for which it can be assumed homogeneous distribution
benzo [a] pyrene in a given lot, it is sufficient to investigate the aggregate sample
take three incremental samples per lot. They must be made to the number
lot. In the case of olive oil and olive pomace oil is
further information on sampling listed in the regulations of the European Communities
^ 30).

As for other products, the minimum number of incremental samples that
to be removed from the lot given in Table 1. The incremental samples shall have
similar weight of not less than 100 g sub-sample,
together and form a bulk sample with a total weight of at least 300 g.
aggregate sample is prepared according to section 3.4.

Table 1:


Minimum number of incremental samples to be taken from the lot
+ -------------------------------- - + ------------------------------- +
| Lot weight (kg) | Minimum number of incremental samples |
| | Be taken |
+ ---------------------------------- + ----------- -------------------- +
| <50 | 3 |
| 50-500 | 5 |
| > 500 | 10 |
+ ---------------------------------- + ----------- -------------------- +

If the lot consists of individual packages, the number of packages subscribed for
to form the aggregate sample given in Table 2.

Table 2:

Number of packages (incremental samples) samples taken in order to create
aggregate sample if the lot consists of individual packages
+ -------------------- ------- + ----------------------------------------- +
| Number of packages or units | Number of packages or units |
| in the lot | to be taken |
+ --------------------------- + ------------------ ----------------------- +
| 1-25 | 1 package or unit |
| 26-100 | about 5%, at least two packages or units |
| > 100 | about 5%, maximum 10 packages or |
| | units |
+ --------------------------- + ------------------ ----------------------- +

4.2 Sampling at retail

Sampling of foodstuffs at retail stage shall be done reasonably
accordance with the provisions of this Annex on sampling. If this is not possible, you can use
effective sampling procedures at retail, they are sampled lot
sufficiently representative.

Fifth Compliance with the specifications of the lot or sublot
control laboratory performs a re-test laboratory sample for confirmation purposes
result, if the result is received by the first test, 20% lower
or greater than the maximum limit, and in such cases, calculate the mean of two results
.

The lot is accepted if it meets the first test result or if it is necessary
retest if it meets the relevant maximum diameter
limit laid down in European Community regulations ^ 30) at
taking into account the measurement uncertainty and correction for recovery.

The lot is rejected if the result of the first test or average, if necessary
retest goes when taking into account the uncertainty
measurement and correction for recovery, the maximum limit specified in Regulation
Commission (EC). 466/2001.


Příl.45
Sample preparation and requirements for methods of analysis used for official
control of benzo [a] pyrene in foodstuffs

First Precautions and general principles for the official control of
benzo [a] pyrene in foodstuffs

Basic requirement is to obtain a representative and homogeneous laboratory
sample without introducing secondary contamination. The analyst must ensure
when preparing samples do not become contaminated. The containers are rinsed before
using acetone or hexane high purity (pa classes
HPLC or equivalent), to minimize the risk of contamination.
Apparatus coming into contact with the sample should be
made of inert materials eg. Aluminum, glass or polished stainless steel
. Not used plastics, such as
polypropylene, polytetrafluoroethylene etc., because they can absorb analytical
sample.

All the collected amount of food received by the laboratory is used to prepare
sample. Reproducible results provide only
thoroughly homogenized samples.

You can also use other methods for preparing samples according to § 1.

Second Treatment of the sample received in the laboratory

Complete aggregate sample is ground finely and mixed thoroughly using a process
which achieves complete homogenisation.

Third Distribution of samples for enforcement and defense purposes

Replicate samples for enforcement, defense
business and referee purposes shall be taken from the homogenised sample.

Fourth Methods of testing and laboratory control requirements
4.1 Definitions

The most common definitions that the laboratory will be used:

r = repeatability, the value below which the absolute
with a certain probability (typically 95%) lie
absolute value of the difference between the results of two separate
specific probability (ie. the same

sample, same operator, same apparatus, same
laboratory determined shortly after each other); r = 2.8 x Sr

Sr = standard deviation, calculated from results
Repeatability.

RSDr = Relative standard deviation calculated from results
_
obtained under repeatability conditions [(Sr / x) x 100]
_
where x is the average of results over all laboratories and samples

R = Reproducibility, the value below which according
expectations with a certain probability (typically 95%)
lie to lie two
separate determinations under the terms
reproducibility (ie.
on identical material obtained by operators in different laboratories, using the standardized test method
); R = 2.8 x SR.

SR = standard deviation calculated from results generated under
reproducibility conditions.

RSDR = relative standard deviation calculated from results
_
generated under reproducibility conditions [(SR / x) x
_
100], where x is the average of results over all laboratories and

samples.

HORRATr = value found RSDr divided by the RSDr value calculated
from the Horwitz equation using the assumption r = 0.66 R.

HORRATR = the observed RSDR value divided by the RSDR value calculated
from the Horwitz equation.

U = the expanded uncertainty, using a factor
cover 2, which corresponds to a reliability level
approximately 95%.
4.2 General requirements


Testing methods used for food control purposes must comply with § 9.

4.3 Specific requirements


If not directly applicable European Community set
specific methods for the determination of benzo [a] pyrene in food laboratories
chooses validated method provided the selected method meets
criteria listed in the table. During validation applies
certified reference material.

Table: Performance characteristics of analytical methods for benzo [a] pyrene
+ ------------------- + ----------- ----------------------------------- +
| Parameter | Value / comment |
+ ------------------- + -------------------------- -------------------- +
| Applicability | Food specified in Regulation (EC) no. |
| | 208/2005 |
+ ------------------- + -------------------------- -------------------- +
| Margin | Less than or equal to 0.3 mcg / kg |
| Detectable | |
+ ------------------- + -------------------------- -------------------- +
| Margin | Less than or equal to 0.9 microg / kg |
| Determination | |
+ ------------------- + -------------------------- -------------------- +
| Accuracy | HORRAT values, or HORRATR achieved in |
| | validation collaborative study must be |
| | less than 1.5 |
+ ------------------- + -------------------------- -------------------- +
| Recovery | 50% to 120% |
+ ------------------- + -------------------------- -------------------- +
| Specificity | Determination must not be interfered with and matrix |
| | spectral phenomena, verification of detection |
+ ------------------- + -------------------------- -------------------- +

4.3.1 Operational characteristics of-concept of uncertainty

Suitability test methods to be used in the laboratory to assess
also use the concept of uncertainty. The laboratory uses a method that
results within the maximum standard uncertainty.
Maximum standard uncertainty is calculated using the following equation:
Uf = square root of (LOD / 2) 2 + (0.2 cm) 2

Where:
Uf is the maximum standard uncertainty
LOD is the limit of detection methods
C is the concentration

If a test method provides results with uncertainty measurements less than the maximum
standard uncertainty, the method will be useful to the same extent as
method which meets the performance characteristics listed in the table.

4.4 Recovery calculation and reporting of results

Analysis results to be reported corrected or uncorrected for recovery.
Must The manner of yield and its value.
Test result corrected for recovery is used for checking compliance.

Be taken into consideration to a European Commission report on the relationship between analytical
results, measurement uncertainty, recovery factors and EC legislation

In the food sector.

The analytical result must be reported as (x +/- U), where x is
analytical result and U is the measurement uncertainty.


Příl.46
Methods of sampling for the official control of the levels of Fusarium toxins in food


First Purpose and scope

Samples for the official control of the levels of Fusarium toxins in food is taken
methods described below. Aggregate samples thus obtained are
considered as representative of the lots.
Compliance with maximum limits laid down in Annex I to Commission Regulation (EC) no. 466/2001 shall
based on the amount determined in the laboratory samples.

Second definition

For the purposes of this Annex batches
identifiable quantity of food delivered at one time and determined by the person referred to in paragraph 3 §
. 1 common characteristics, such as origin, variety, type of packaging
packer, consignor or markings.

Third General provisions

3.1 Material to be removed

Each lot which is to be examined must be sampled separately.
Large lots in item 4.3 must be subdivided into sublots to be sampled separately
.

3.2 Precautions

Of sampling and sample preparation must be done
precautions to avoid any changes which would affect the Fusarium toxin content
, adversely affect the analytical determination or
aggregate samples unrepresentative.
Sub-samples 3.3


Incremental samples shall, if possible, from various places throughout the lot or sublot
.

Deviation from this procedure must be recorded in the log.

3.4 Preparation of aggregate sample

The aggregate sample is made up by uniting the incremental samples.
3.5 Replicate samples


Replicate samples for enforcement, defense
commercial dispute or arbitration shall be taken from the homogenised aggregate sample
.

3.6 Packaging and transport of samples

Each sample is placed in a clean, inert container which
provides adequate protection from contamination and damage during transportation
. They must be taken all the necessary measures to prevent
change in composition of the sample, which might arise during transportation or storage.

3.7 Sealing and labeling of samples

Each sample taken for official use shall be sealed at the place of sampling and identified following
§ 6

Of each sampling record must be kept for sampling by
§ fifth

Fourth Special provisions

4.1 Different types of lots

The food is put into circulation in bulk, containers, or
individual packs, such as sacks, bags or individual
retail packaging. Sampling is performed for each kind of putting into circulation
.

Without prejudice to the specific provisions of paragraphs 4.3, 4.4 and 4.5 below
formula can be used as a guide for the sampling of lots
have when put into circulation a form of individual packs, such as sacks, bags || | retail packings.
weight of the lot x weight of the incremental sample
sample size n = ------------------------------------- --------------------
Weight of the aggregate sample x weight of each package

- Weight - in kg
- Sample size - every nth sack or bag from which
They must be removed subsample (
decimal places are rounded to the nearest whole number
).

4.2 Weight of the incremental sample

Weight of the incremental sample shall be at least 100 grams, unless this
Annex provides otherwise. For lots in retail packing
depends on the weight of the incremental sample weight of the retail pack.

4.3 Sampling procedure for cereals and cereal products

Table 1: Subdivision of lots into sublots depending on product and lot weight

-------------------------- --------------------------------------------------
Commodity lot weight Weight or number of weight
(T) the number of split samples aggregate
batch of sample (kg)
------------------------------------------ ----------------------------------
Cereals> = t 100 1500 500 10
and products -------------------------------------------------- ---------------
cereal> 300 and <1 500 3 sublots 100 10

-------------------------------------------------- ---------------
> = 50 and = <300100 tons 100 10
-------------------------------------------------- ---------------
<50 - 3-100 *) 1-10
------------------------------------ ----------------------------------------
*) depending on the lot weight - see table 2 -------------------------------------------
---------------------------------

4.4 Sampling procedure for cereals and cereal products for lots of 50 tonnes, including


Sublot may be separated physically, each lot must be physically
divided into sublots following Table 1. Given the weight
lot is not always an exact multiple of the weight of the lot, the weight of
batch exceed the mentioned weight by a maximum of 20%.

Each sublot must be sampled separately.

If it is not possible to use the method of sampling set out in this point because of
economic consequences resulting from damage to the lot
such as packaging or mode of transport may be used
alternative method of sampling provided that is what
representative as possible and is fully described and documented.

4.5 Sampling procedure for cereals and cereal products for lots of 50 tonnes


For lots of cereals and cereal products less than 50 tonnes must be
depending on the weight of the lot used in the sampling plan consisting of 10 to 100
incremental samples resulting in an aggregate sample of 1 to 10 kg.
For very small lots (= <0,5 tonnes) may be taken
lower number of incremental samples, but the aggregate sample uniting all incremental samples shall be also
in that case at least 1 kg.

The figures in Table 2 are used to determine the number of incremental samples
to be taken.

Table 2: Number of incremental samples to be taken depending on the weight
lot of cereals and cereal products
------------------- -------------------------------
lot weight (tonnes) Number of incremental samples
----- ---------------------------------------------
= <
0.05 3> 0.05 - = <0.50 5
> 0,50 - = <1.00 10
> 1,00 - = <3.00 20
> 3.00 - = <40
10.00> 10.00 - = <60
20.00> 20.00 - = <50.00 100 ---------
-----------------------------------------


4.6 The sampling procedure for foods intended for infants and young children


For foods intended for infants and young children, the procedure of collection of samples
mentioned for cereals and cereal products in 4.5. Number
incremental samples to be taken depends on the weight of the lot,
according to Table 2, a minimum of 10 and maximum of 100 sub-samples.
For very small lots (= <0,5 tonnes) may be taken
lower number of incremental samples, but the aggregate sample uniting all incremental samples shall be also
in that case at least 1 kg.

Weight of the incremental sample shall be at least 100 grams. For lots in retail packing
depends on the weight of the incremental sample weight
packaging business and in very small lots (= <0,5 tonnes) must have
sub-samples of such weight that uniting the incremental samples originated summary || | specimen weighing at least 1 kg.

The aggregate sample is 1-10 kg and the sample must be sufficiently
mixed.

4.7 Sampling at retail

Sampling of foodstuffs at retail stage shall be done reasonably
accordance with the provisions of this Annex on sampling. If this is not possible, you can
use other effective sampling procedures at retail
if the sampled lot sufficiently representative.

Fifth Acceptance of a lot or sublot

The lot is accepted if the aggregate sample conforms to the maximum limit
taking into account the measurement uncertainty and correction for recovery.

The lot is rejected if the aggregate sample exceeds any doubt
maximum limit, taking into account the measurement uncertainty and correction for
recovery.


Příl.47
Sample preparation and requirements for methods of analysis used for official
control of the levels of Fusarium toxins in food

First Precautionary measures

As the distribution of Fusarium toxins is very uneven
must prepare laboratory specimens and in particular

Samples are homogenized given special attention. All collected
quantity of food is used to prepare the test sample.

Second Treatment of the sample received in the laboratory

Each laboratory sample finely grinded and mixed thoroughly process
which achieves complete homogenisation.

If the maximum level applies to the solids content is determined
product of the homogenised sample, using a method that proved
to determine accurately the dry matter.

Third Distribution of samples for testing to confirm the result, and the defense


Replicate samples for enforcement, defense
business and referee purposes shall be taken from the homogenised sample
.

Fourth Methods of testing and laboratory control requirements
4.1 Definitions


The most common definitions that the laboratory will be used:

r = repeatability, the value below which will by
expectations with a certain probability (typically 95%
) lie to lie 2
separate determinations under the terms
Repeatability (ie. the same sample, same
operator, same apparatus, same laboratory,
determined in quick succession); r = 2.8 x sr,
sr = standard deviation, calculated from results
obtained under repeatability conditions.
RSDr = relative standard deviation calculated from
the results obtained under the given conditions
-
Repeatability [(s / Rx) x 100]
R = Reproducibility, the value below which according
expectations with a certain probability (typically 95%
) lie to lie
two separate determinations under the terms
reproducibility (ie.
on identical material obtained by operators in different laboratories, as
using the standardized test method); R =
2.8 x sR
Standard deviation, calculated from results
under reproducibility conditions,
RSDR = relative standard deviation calculated from
the results obtained under the conditions
-
Reproducibility [(sR / x) x 100].
4.2 General requirements


Testing methods used for food control purposes must comply with § 9.

4.3 Specific requirements


4.3.1 Performance characteristics

If not directly applicable European Community set
special methods, laboratory chooses validated method provided the
selected method meets the performance characteristics given in Tables 1
2, 3 and 4. || | Table 1: Performance characteristics for deoxynivalenol

----------------------------------------------- -------------------
amount of microg / kg deoxynivalenol
-------------------------------------------
RSDr% RSDR% Recovery% -----------------------------------------
-------------------------
> 100.00 - = <= 500.00 <= 20.00 <40.00 60 00 - 110.00
------------------------------------------ ------------------------
> = 500.00 <= 20.00 <40.00 70.00 to 120.00 || | ------------------------------------------------- -----------------

Table 2: Performance characteristics for zearalenone

----------------------------------------------- -------------------
amount of microg / kg zearalenone
-------------------------------------------
RSDr% RSDR% Recovery% -----------------------------------------

= ------------------------- <= 50.00 <= 40.00 <50.00 60.00 to 120.00
----------------------------------------------- -------------------
> 50.00 = <= 25.00 <40.00 70.00 to 120.00
---- -------------------------------------------------- ------------

Table 3: Performance characteristics for fumonisin B1 and B2

----------------------------------------------- -------------------
amount of micro / kg Fumosin B1 or B2
-------------------------------------------
RSDr% RSDR% Recovery% -----------------------------------------

= ------------------------- <= 500.00 <= 30.00 <60.00 60.00 to 120.00
----------------------------------------------- -------------------
> = 500.00 <= 20.00 <30.00 70.00 to 110.00
---- -------------------------------------------------- ------------

Table 4: Performance characteristics for T-2 and HT-2 toxin

----------------------------------------------- -------------------

Quantity of micro / kg T-2 toxin
-------------------------------------------
RSDr% RSDR% Recovery% -----------------------------------------
-------------------------
50.00 to 250.00 = <40.00 = <60,00 60,00 - 130 00
--------------------------------------------- ---------------------
> = 250.00 <= 30.00 <50.00 70.00 to 130.00
- -------------------------------------------------- --------------

----------------------------------------------- -------------------
amount of microg / kg HT-2 toxin
-------------------------------------------
RSDr% RSDR% Recovery% -----------------------------------------
-------------------------
100.00 to 200.00 = <40.00 = <60,00 60,00 - 130 00
--------------------------------------------- ---------------------
> = 200.00 <= 30.00 <50.00 70.00 to 130.00
- -------------------------------------------------- --------------

Detection limits of the methods used are not stated as the precision
given the concentrations.

Accuracy of the method corresponds to the value calculated from the Horwitz equation:

RSDR = 2 (1 - 0,5 log C)


Where:

- RSDR is the relative standard deviation calculated from
the results obtained under reproducibility conditions
-
[SR / x) x 100]
- C is the concentration ratio (i.e. 1 = 100 g / 100 g, 0.001%
1000 mg / kg).


This is a generalized equation for the precision with which it was shown that in most routine methods of analysis
independent of analyte and matrix but only
concentration.

4.3.2 An approach based on suitability for purpose

To evaluate the acceptability of methods of analysis may be used
alternative approach to fitness for a particular purpose, within which define
only fitness function as a single parameter, if there
limited number of fully validated test methods. A fitness function is the function
uncertainty that specifies maximum levels of uncertainty regarded
as fit for purpose.

Given the limited number of test methods that are fully validated
collaborative trial, especially for the determination of T-2 and HT-2 toxin, is used
function approach uncertainties fixing the maximum
acceptable uncertainty, also to assess the suitability for a particular purpose
methods of testing lab uses.
The laboratory may use a method which produces results within the maximum standard uncertainty.
The maximum standard uncertainty is calculated using the following equation:

Uf = root (LOD / 2) 2 + (alpha x C) 2


Where:
Uf is the maximum standard uncertainty
LOD is the limit of detection methods
C is the concentration (mcg / kg)
alpha is a constant, numeric factor to be used
Depending on the value of C. The values ​​to be
used are shown in Table 5.

If a test method provides results with uncertainty measurements less than the maximum
standard uncertainty, the method will be useful to the same extent as
method which meets the performance characteristics given in Tables
.
Table 5: Numeric values ​​to be dependent upon
the concentration of alpha used for as constant in
formula set out in this point

----------------------------------------------- ---
C (microg / kg) alpha
---------------------------------- ---------------- =
<
50.00 0.20 -------------------- ------------------------------
51.00 to 500.00 0.18
---- ---------------------------------------------- 501
00 - 1000.00 0.15 --------------------------------------
------------
1001.00 to 10000.00 0.12
---------------------- ----------------------------
>
10,000.00 0.10 --------- -----------------------------------------

4.4 Recovery calculation and reporting of results

The test results are reported corrected or uncorrected for recovery.
Must The manner of yield and its value.
Test result corrected for recovery is used for checking compliance.

The test result must be presented in the form (x +/- U), where x is the result
and U is the expanded measurement uncertainty.

4.5 Requirements for laboratories

Laboratories must comply with the special regulation. ^ *)

1) Issued on the basis and within the law, which allows content
incorporate the relevant European Community regulations by decree.


1) First Commission Directive 79/1067 / EEC of 13 November 1979 amending
establishing analytical methods for testing certain
dehydrated preserved milk for human consumption.

First Commission Directive 79/796 / EEC of 26 July 1979 laying down
Community methods of analysis for testing certain sugars
intended for human consumption.

Commission Directive 80/891 / EEC of 25 July 1980 concerning
Community methods of analysis for determining the erucic acid content in
oils and fats intended as such for human consumption and in foodstuffs containing added
oils or fats.

First Commission Directive 81/712 / EEC of 28 July 1981 laying down
Community methods for verifying compliance with the criteria for purity
certain additives used in food.

First Commission Directive 85/503 / EEC of 25 October 1985 on methods of analysis
edible caseins and caseinates.

Council Directive 85/591 / EEC of 20 December 1985 concerning the introduction
Community methods of sampling and analysis for the monitoring of food
intended for human consumption.

First Commission Directive 86/424 / EEC of 15 July 1986 laying down
methods of sampling for chemical analysis of edible caseins and caseinates
.

First Commission Directive 87/524 / EEC of 6 October 1987 laying down
Community methods of sampling for chemical analysis
monitored dairy products.

Commission Directive 92/2 / EEC of 13 January 1992 laying down
sampling procedure and the methods of analysis at Community
official control over the temperatures of frozen foods intended for human consumption.

Council Directive 93/99 / EEC of 29 October 1993 on additional measures
concerning the official control of foodstuffs.

Commission Directive 98/53 / EC of 16 July 1998 laying down
methods of sampling and analysis for the official control of the levels of certain
contaminants in food.

Commission Directive 2001/22 / EC of 8 March 2001 laying down
methods of sampling and analysis for the official control of
levels of lead, cadmium, mercury and 3-MCPD in foods.

Directive of the European Parliament and Council Directive 2001/37 / EC of 5 June 2001 on the approximation
regulations and administrative provisions of the Member States relating
the manufacture, presentation and sale of tobacco products.

Commission Directive 2002/26 / EC of 13 March 2002 laying down
methods of sampling and analysis for the official control of
ochratoxin A in food.

Commission Directive 2002/27 / EC of 13 March 2002 amending
Directive 98/53 / EC laying down the sampling methods and the methods
analysis for the official control of the levels for certain contaminants in
foods.

Commission Directive 2002/63 / EC of 11 July 2002 laying down
Community methods of sampling for the official control
residues of pesticides in products of plant and animal origin and on their surface and
repealing Directive 79/700 / EEC.

Commission Directive 2002/69 / EC of 26 July 2002 laying down
methods of sampling and analysis for the official control of dioxins and
determination of dioxin-like PCBs in food.

Commission Directive 2003/78 / EC of 11 August 2003 laying down
methods of sampling and analysis for the official control of the levels of patulin in foodstuffs
.

Commission Directive 2003/121 / EC of 15 December 2003 amending
Directive 98/53 / EC laying down the sampling methods and the methods
analysis for the official control of the levels for certain contaminants in
foods.

Directive of the European Parliament and Council Directive 2003/114 / EC of 22 December
2003 amending Directive 95/2 / EC on food additives
additives other than colors and sweeteners.

Commission Directive 2004/16 / EC of 12 February 2004 laying down
methods of sampling and analysis for the official control of tin in
foods packaged in cans.

Commission Directive 2004/43 / EC of 13 April 2004 amending
Directive 98/53 / EC and 2002/26 / EC as regards the methods of sampling and analysis for the
official control levels of aflatoxin and ochratoxin a
in foods for infants and young children.

Commission Directive 2004/44 / EC of 13 April 2004 amending

Directive 2002/69 / EC laying down the sampling methods and
analytical methods for official control of dioxins and the determination of dioxin-like PCBs in
food.

Commission Directive 2005/4 / EC of 19 January 2005 amending
Directive 2001/22 / EC laying down the sampling methods and the methods
analysis for the official control of the levels of lead,
cadmium, mercury and 3-MCPD in foods.

Commission Directive 2005/5 / EC of 26 January 2005 amending
Directive 2002/26 / EC laying down the sampling methods and the methods
analysis for the official control of the levels of ochratoxin A in foods .

Commission Directive 2005/10 / EC of 4 February 2005 laying down
methods of sampling and analysis for the official control of
benzo [a] pyrene in foodstuffs.

Commission Directive 2005/38 / EC of 6 June 2005 laying down
methods of sampling and analysis for the official control of
Fusarium toxins in food.

2) Act no. 110/1997 Coll., On foodstuffs and tobacco products and
amending and supplementing some related laws, as amended
regulations.

3) For example, Act no. 146/2002 Coll., On the State Agricultural and Food Inspection
and amending certain related acts as amended by Act no. 309/2002 Coll
. and Act no. 94/2004 Coll. Act no. 166/1999 Coll., On
veterinary care and amending certain related laws (Veterinary
Act), as amended by Act no. 29/2000 Coll., Act no. 154/2000 Coll., Act
No. 102/2001 Coll., Act no. 76/2002 Coll., Act no. 120/2002 Coll., Act No.
. 309/2002 Coll., Act no. 320/2002 Coll. and Act no. 131/2003 Coll.

4) ISO 3951 Acceptance procedures and charts for inspection by variables for percent nonconforming
units.

ISO 2859-0 Sampling procedures for inspection by attributes. Part 0: Introduction
takeovers of attributes ISO 2859.

ISO 2859-1 Sampling procedures for inspection by attributes. Part 1: Acceptance
AQL plans to check every batch in the series.

ISO 2859-2 Sampling procedures for inspection by attributes. Part 2: Sampling plans
LQ for isolated lot inspection.

ISO 2859-3 Sampling procedures for inspection by attributes. Part 3: Intermittent
acceptance.

ISO 2859-4 Part 4: Procedures for assessing
specified levels of quality.

ISO 10725 Selective sampling plans and procedures to check
bulk materials.

5) ISO 8243 Cigarettes. Sampling.

ISO 4874 Tobacco. Sampling of raw material batch.

ISO 15592-1 Fine-cut tobacco and tobacco products piece intended to
smoke produced from it. Methods of sampling, conditioning and analysis.
Part 1: Sampling.

6) CSN 56 0003 Sampling and testing methods for determination of aflatoxins in foods
.

CSN 56 0253 Sampling for the determination of pesticides in and on fruit and vegetables
.

ISO 1839 Tea. Sampling.

ISO 13690 Cereals, pulses and milled products. Sampling of statistical
benefits.

EN ISO 5555 Animal and vegetable fats and oils. Sampling.

CSN 56 0290-2 Methods of test for frozen products. Part 2: Sampling.

7) CSN 57 0105-2 Methods of testing of dairy products and dried
thickened. Part 2: Sampling.

CSN 57 0111-1 Methods of test for casein and caseinates. Part 1: General
provisions.

CSN 57 0111-2 Methods of test for casein and caseinates. Part 2: Sampling for chemical analysis
.

8) Commission Decision no. 91/180 / EEC of 14 February 1991
down certain methods of analysis and testing of raw milk and heat-treated milk
.

9) Commission Regulation (EC) no. 1148/2001 of 12 June 2001 on
checks compliance with marketing standards for fresh fruit and vegetables.

10) Commission Regulation (EC) no. 2568/1991 of 11 July 1991 on
characteristics of olive oil and remaining olive oil and
relevant methods of analysis, as amended by Commission Regulation (EEC) No. | || 3682/1991, Commission Regulation (EC) no. 1429/1992 and Commission Regulation (EEC) No.
1683/1992, Commission Regulation (EC) no. 3288/1992 and Commission Regulation (EEC) No. | || 183/1993, Commission Regulation (EC) no. 177/1994, Commission Regulation (EC) no. 656/1995
Commission Regulation (EC) no. 2527/1995, Commission Regulation (EC) no. | || 2472/1997, Commission Regulation (EC) no. 282/1998, Commission Regulation (EC) no. 2248/1998
Commission Regulation (EC) no. 379/1999, Commission Regulation (EC) No.

2042/2001 and Commission Regulation (EC) no. 796/2002.

10a) Commission Recommendation 2004/787 / EC of 4 October 2004 on technical
guidance for sampling and detection of genetically modified organisms
and material produced from genetically modified organisms or their products
contents under Commission Regulation (EC) no. 1830/2003.

11) Decree no. 38/2001 Coll., On hygienic requirements on products
intended for contact with food and dishes, as amended by Decree no. 186/2003 Coll
.

12) EN ISO 661 Animal and vegetable fats and oils.
Sample preparation for analysis.

13) ISO 5496 Sensory Analysis. Methodology. Initiation into
and training of assessors in the detection and recognition of odors.

ISO 8586-1 Sensory analysis. General guidance for selection, training and monitoring activities
assessors. Part 1: Selected assessors.

ISO 8586-2: Sensory Analysis. General guidance for selection, training and monitoring activities
assessors. Part 2: Experts.

14) ISO 8589 Sensory Analysis. General guidelines for the layout
sensory workplace.

ISO 11035 Sensory analysis. Identification and selection of descriptors for determining
sensory profile using a multidimensional approach.

ISO 8587 Sensory Analysis. Methodology. Serial test.

ISO 11036 Sensory analysis. Methodology. Profile texture.

ISO 11037 Sensory analysis. General guidelines and test method for assessing
food colors.

ISO 11056 Sensory analysis. Methodology. Method content of magnitude.

ISO 8588 Sensory Analysis. Methodology. The test "A" - not "A".

ISO 3972 Sensory Analysis. Methodology.
Method of studying the sensitivity of taste.

15) ISO / IEC Directives, Part 2, 2001, 4th edition (Rules for the structure and drafting international standards
).

16)

ISO 5725-1 Accuracy (trueness and precision) of measurement methods and results.
Part 1: General principles and definitions.

ISO 5725-2 Accuracy (trueness and precision) of measurement methods and results.
Part 2: Basic method for the determination of repeatability and reproducibility
standard measurement method.

ISO 5725-3 Accuracy (trueness and precision) of measurement methods and results.
Part 3: Intermediate measures of precision standard measurement method.

ISO 5725-4 Accuracy (trueness and precision) of measurement methods and results.
Part 4: Basic methods for the determination of the correctness of standardized methods for measuring
.

ISO 5725-5 Accuracy (trueness and precision) of measurement methods and results.
Part 5: Alternative methods for determining the precision
standardized methods.

ISO 5725-6 Accuracy (trueness and precision) of measurement methods and results.
Part 6: Using the values ​​of precision measurements in practice.

17)

EN ISO / IEC 17025 General requirements for the competence of testing and calibration laboratories
.

§ 16 of Act no. 22/1997 Coll., On technical requirements for products and
amending and supplementing certain acts as amended by Act no. 71/2000 Coll.
Act no. 102/2001 Coll . and Act no. 205/2002 Coll.

18) ISO 8243 Cigarettes. Sampling.

ISO 8454 Cigarettes. Determination of carbon monoxide in smoke condensates
. Method mdir.

ISO 10315 Cigarettes. Determination of nicotine in smoke condensates
. Gas chromatographic method.

ISO 4387 Cigarettes. Determination of crude and condensate beznikotinového
smoke using routine analytical nakuřovacího device.

19) CSN 57 0111-1 Methods of test for casein and caseinates. Part 1: General Provisions
.

CSN 57 0111-3 Methods of test for casein and caseinates. Part 3: Determination
moisture.

CSN 57 0111-5 Methods of test for casein and caseinates. Part 5:
protein content.

CSN 57 0111-7 Methods of test for casein and caseinates. Part 7: Determination
ash content.

CSN 57 0111-8 Methods of test for casein and caseinates. Part 8: Determination
titratable acidity.

CSN 57 0111-12 Methods of testing casein and caseinates. Part 12: Determination
pH.

20) CSN 57 0105-3 Methods of testing of dairy products and dried
thickened. Part 3: Determination of solids in sweetened and unsweetened milk
.

CSN 57 0111-10 Methods of testing of dairy products, powdered and condensed.
Part 10: Determination of lactic acid and lactates.

CSN 57 0111-11 Methods of testing of dairy products, powdered and condensed.

Part 11: Determination of phosphatase activity in powdered milk.

CSN 57 0111-13 Methods of testing of dairy products, powdered and condensed.
Part 13: Determination of water content in powdered milk.

21) CSN 57 0290-7 Methods of test for frozen products. Part 7: Measurement
temperatures.

Decree no. 61/1983 Coll., On the Agreement on the International Carriage of Perishable Foodstuffs and
special equipment used for
such Carriage (ATP), as amended.

22) Commission Regulation (EC) no. 2870/2000 of 19 December 2000 laying down
Community reference methods for the analysis of spirit drinks, as
Regulation (EC) no. 2091/2002.

23) Commission Regulation (EC) no. 558/1993 of 10 March 1993
refractometry method of measuring dry soluble residue in products
processed fruit and vegetables, repealing Regulation (EEC) No .
and 543/1986 amending Annex I to Council Regulation (EC) no. 2658/1987.

24) Commission Regulation (EC) no. 4154/1987 of 22 December 1987 laying down
methods of analysis and other technical provisions necessary for
implementation of Regulation (EC) no. 3033/1980, establishing business
measures applicable to certain goods resulting from
processing of agricultural products, as amended by Commission Regulation (EC) no. 203/1998
.

25) of Commission Regulation (EC) no. 213/2001 of 9 January 2001
rules for implementing Council Regulation (EC) no. 1255/1999 as regards methods for the analysis
and quality evaluation of milk and milk products and laying
amending Regulation (EC) no. 2771/1999 and (EC) no. 2779/1999.

26) Act no. 115/1995 Coll., On viticulture and viniculture and amending and supplementing certain
related legislation, as amended by Act no. 216/2000 Coll
., Law no. 50/2002 Coll. and Act no. 147/2002 Coll.

27) CSN 01 1300 Quantities and units.

ISO 1000 SI units and recommendations for the use of their multiples and
of certain other units.

28) EN ISO / IEC 17025 General requirements for the competence of testing and calibration laboratories
.

29) Decree no. 54/2004 Coll., On foodstuffs intended for particular nutritional uses
and method of their use.

30) Commission Regulation (EC) no. 1989/2003 of 6 November 2003
amending Regulation EEC no. 2568/91 on the characteristics of olive oil and
olive pomace oil and on the relevant methods analysis.

*) For example, § 3 para. 3 of the Act no. 146/2002 Coll., On State Agricultural and Food Inspection
and amending certain related laws, as amended
.