BETWEEN THE PROVISIONS OF LAW 25/1970 OF 2 DECEMBER, WHICH APPROVED THE STATUS OF VINEYARD, WINE AND SPIRITS, PROVIDED FOR IN ARTICLE 34.2, THAT COMPOUND SPIRITS AND LIQUEURS AND, AMONG THEM, ANISE, WOULD BE SUBJECT OF SPECIAL REGULATIONS.

AS A CONSEQUENCE OF THE PREVIOUS FORECAST WAS PUBLISHED THE ROYAL DECREE 644/1982 OF 5 MARCH (2 ), WHICH APPROVED THE SPECIAL REGULATIONS FOR THE ELABORATION, CIRCULATION AND TRADE OF ANISE, IT IS NOW NECESSARY TO REGULATE ITS ANALYTICAL METHODS.

IN THE DRAFTING OF OFFICIAL METHODS OF ANALYSIS, IT HAS BEEN SOUGHT, AS FAR AS POSSIBLE AND IN ACCORDANCE WITH THE GUIDELINES SET OUT IN ARTICLE 9. ROYAL DECREE 1908/1984 OF 26 SEPTEMBER ( OF 29 OCTOBER) AMENDING CERTAIN OF THE ARTICLES AND HEADINGS OF CERTAIN REGULATIONS FOR THE PRODUCTION, CIRCULATION AND TRADE IN BEVERAGES DERIVED FROM NATURAL ALCOHOLS, THEIR ADAPTATION TO THE METHODS APPROVED BY THE INTERNATIONAL ORGANIZATIONS SPECIALIZED IN THE FIELD, IN ORDER TO TAKE ADVANTAGE OF THE EXPERIENCES OBTAINED FROM THEIR APPLICATION.

IN ITS VIRTUE, ON THE PROPOSAL OF THE MINISTRIES OF ECONOMY AND FINANCE, OF INDUSTRY AND ENERGY, OF AGRICULTURE, FISHERIES AND FOOD AND OF HEALTH AND CONSUMPTION, THE REPRESENTATIVES OF THE SECTORS AFFECTED, AND PREVIOUS PRECEPT REPORT OF THE INTERMINISTERIAL COMMISSION FOR FOOD MANAGEMENT, THIS MINISTRY OF RELATIONS WITH THE COURTS AND THE SECRETARIAT OF GOVERNMENT HAS:

FIRST. THE METHODS OF ANALYSIS FOR THE ANISE REFERRED TO IN ANNEX 1 ARE APPROVED AS OFFICIAL.

SECOND. ANALYTICAL DETERMINATIONS SHALL BE CARRIED OUT IN ACCORDANCE WITH THE OFFICIAL METHODS IN FORCE. WHERE THERE ARE NO OFFICIAL METHODS FOR CERTAIN ANALYSES AND UNTIL SUCH METHODS ARE PROPOSED BY THE COMPETENT AUTHORITY AND NOTIFIED IN ADVANCE BY THE INTER-MINISTERIAL COMMISSION FOR FOOD MANAGEMENT, THE FOLLOWING MAY BE USED: APPROVED BY THE NATIONAL AND INTERNATIONAL BODIES OF RECOGNISED SOLVENCY.

REPEAL PROVISION

AS MANY PROVISIONS OF EQUAL OR LOWER RANK ARE REPEALED, THEY SHALL OPPOSE THE PROVISIONS OF THIS ORDER.

FINAL DISPOSITION

THIS ORDER SHALL ENTER INTO FORCE WITHIN SIX MONTHS OF ITS PUBLICATION IN , EXCEPT FOR METHODS NUMBER 6, ANETOL, AND NUMBER 9, ESSENTIAL OILS, WHICH SHALL ENTER INTO FORCE WITHIN 18 MONTHS OF PUBLICATION. IN THE .

MADRID, 8 MAY 1987.

ZAPATERO GOMEZ

EXCMOS. SRES. MINISTERS FOR HEALTH AND CONSUMER AFFAIRS, ECONOMY AND FINANCE, INDUSTRY AND ENERGY, AGRICULTURE, FISHERIES AND FOOD.

ANNEX

1.

OBTAINING FROM THE TARGET.

2.

SPECIFIC WEIGHT OF THE NATURAL ESSENTIAL OIL (IN RAW MATERIAL).

3.

MELTING POINT OF NATURAL ESSENTIAL OIL (IN RAW MATERIAL).

4.

ALCOHOLIC STRENGTH.

5.

TOTAL SUGARS.

6.

ANETOL.

7.

METHANOL.

8.

NITROGENOUS BASES.

9. ESSENTIAL OILS.

10.

FURFURAL.

11.

ARSENIC.

12.

LEAD.

13.

CINC.

14.

COBRE.

1. GETTING THE TARGET

1.1 PRINCIPLE

DISTILLATION OF THE SAMPLE UNDER CERTAIN CONDITIONS.

1.2 MATERIAL AND APARATOS

1.2.1 FLASK OF 500 TO 1,000 MILLILITRES OF CAPACITY, OF ROUND BOTTOM AND GRINDING MOUTH.

1.2.2 KJELDAHL OR SIMILAR ELONGATION.

1.2.3 DIMROTH TYPE REFRIGERANT, 20-30 CENTIMETERS IN LENGTH OR SIMILAR.

1.2.4 VOLUMETRIC 100-ML CAPACITY FLASKS.

1.2.5 ELECTRIC BLANKET OR GAS MECHANIC.

1.3 REAGENTS

1.3.1 WATER-TARGET WATER.

1.4 PROCEDURE

1.4.1 SAMPLES WITH LESS THAN 60 PER 100 ETHANOL. MEASURE 200 MILLILITRES OF SAMPLE IN A GRADUATED FLASK THAT BEFORE ENRASAR IS KEPT IN WATER BATH AT 20 DEGREES CELSIUS FOR HALF AN HOUR. MAKE UP WITH PIPETTE, CLEANING THE INSIDE OF THE NECK OF THE FLASK WITH FILTER PAPER OF POSSIBLE DROPS ATTACHED. AIR BUBBLES SHOULD BE PREVENTED FROM BEING EXPOSED TO THE WALLS OF THE FLASK. QUANTITATIVELY PASS THE MEASURED VOLUME TO THE DISTILLATION FLASK, WASHING THREE CONSECUTIVE TIMES THE VOLUMETRIC FLASK OF 200 MILLILITRES WITH 20 MILLILITRES OF DISTILLED WATER EACH TIME, ADDING THE WASHING WATERS TO THE DISTILLATION FLASK. WHEN IT COMES TO SIRUPOUS DRINKS, DUE TO ITS HIGH SUGAR CONTENT, THE WASHING MUST BE DONE CONSECUTIVELY WITH THREE SERVINGS OF 50 MILLILITRES OF DISTILLED WATER.

PROCEED TO DISTILLATION.

IT IS CONVENIENT TO ADD GLASS BALLS TO FACILITATE SMOOTH BOILING.

IN SOME CASES, IF FOAMING IS FEARED, ADD AN INERT ANTIFOAMING AGENT (SUCH AS SILICONE).

THE DISTILLATE COLLECTION FLASK IS THE SAME AS THE MEASURE. IT IS ADDED 10 MILLILITRES OF DISTILLED WATER AND IS SLIGHTLY INCLINED, SO THAT THE DISTILLATE SLIPS WITHOUT SPLASHES THROUGH THE WALL OF THE FLASK, WHICH MUST BE INSIDE AN ICE BATH OR WATER AT LESS THAN 1 DEGREE CELSIUS.

THE REFRIGERANT MUST BE FITTED WITH AN ELONGATION WHICH PENETRATES AT LEAST 4-5 CENTIMETRES INTO THE NECK OF THE FLASK.

WHEN ABOUT 190 MILLILITRES HAVE BEEN COLLECTED, THE WATER BATH HAS BEEN PLUGGED INTO THE WATER BATH AT 20 DEGREES CELSIUS.

WHEN THE TEMPERATURE IS REACHED, WITHIN THE SAME BATHROOM, MAKE UP TO THE WATER DISTILLED BY PIPETTE.

TAAPAR THE FLASK AND INVERT IT SEVERAL TIMES TO HOMOGENIZE ITS CONTENTS BEFORE PROCEEDING TO THE ETHANOL DETERMINATION.

1.4.2 SAMPLES WITH MORE THAN 60 PER 100 ETHANOL.

THE PREVIOUSLY DESCRIBED PROCEDURE DIFFERS ONLY IN THAT THE SAMPLE IS MEASURED IN A 100 MILLILITRE GRADUATED FLASK, WHICH IS WASHED SEVERAL TIMES WITH WATER UNTIL IT REACHES A VOLUME OF APPROXIMATELY 230 MILLILITRES. THE DISTILLATE IS COLLECTED IN VOLUMETRIC FLASK OF 200 MILLILITRES AND IS WRAPPED WITH DISTILLED WATER INSIDE THE WATER BATH AT 20 DEGREES CELSIUS.

1.5 REMARKS

1.5.1 THE DISTILLATION SYSTEM SHOULD BE CHECKED IN BOTH CASES AS FOLLOWS:

DISTILL 200 MILLILITRES FROM A HYDROALCOHOLIC MIXTURE TO 10 PER 100 BY VOLUME FIVE TIMES SUCCESSIVE. DETERMINE IN THE LAST DISTILLATION THE ALCOHOL DEGREE OF DISTILLATE WHICH MUST NOT BE LESS THAN 9.9 PER 100 IN VOLUME OF ETHANOL.

1.5.2 DISTILLATION MUST BE CARRIED OUT BY ENSURING THAT THE DISTILLATE FLOW IS UNIFORM.

1.5.3 THE DISTILLATE COLLECTION TIME SHALL BE BETWEEN 45 AND 90 MINUTES.

1.5.4 THE DISTILLATE MAY HAVE A MILKY APPEARANCE AS A RESULT OF THE EMULSION OF ESSENTIAL OILS.

1.6 REFERENCES

1.

OFFICIAL METHODS OF ANALYSIS OF RUM, GIN AND WHISKY.

2. SPECIFIC WEIGHT OF NATURAL ESSENTIAL OIL (RAW MATERIAL) 2.1 PRINCIPLE

IS BASED ON THE DETERMINATION OF THE RELATIVE DENSITY OF THE ESSENTIAL OIL WITH RESPECT TO WATER AT THE SAME TEMPERATURE.

THIS DENSITY RELATIVE TO 25 DEGREES CELSIUS WILL BE DETERMINED TO AVOID POSSIBLE SOLIDIFICATION OF THE SAMPLE.

2.2 MATERIAL AND APARATOS

2.2.1 THERMOSTATIC WATER BATH AT 25 0.1 DEGREES CELSIUS.

2.2.2 PICNOMETER OF 50 MILLILITRES, AS ATTACHED FIGURE (FIG. 2.1).

2.2.3 THERMOMETER OF 10 DEGREES CELSIUS 30 DEGREES CELSIUS, GRADUATED IN TENTHS OF A DEGREE.

2.2.4 CAPILLARY TUBE.

2.2.5 ANALYTICAL BALANCE.

2.3 PROCEDURE

2.3.1 COMPLETELY FILL IN THE PICNOMETER WITH RECENTLY DISTILLED WATER, PLUG AND PLACE IN 2.2.1, SO THAT THE WATER LEVEL OF THE BATH IS SLIGHTLY ABOVE THE LEVEL OF ROOTS OF THE PICNOMETER.

2.3.2 AFTER THIRTY MINUTES REMOVE THE STOPPER AND WITH 2.2.4 ADJUST THE WATER LEVEL UNTIL THE LOWER PART OF THE MENISCUS IS TANGENT WITH THE ROOT SIGNAL OF THE PICNOMETER.

2.3.3 REMOVE THE PICNOMETER FROM THE BATHROOM. WITH A SMALL ROLL OF FILTER PAPER DRY THE INSIDE OF THE NECK OF THE PICNOMETER AND THE STOPPER. ENTER EN BATH AT ROOM TEMPERATURE FOR FIFTEEN MINUTES TO SPEED UP THE LIQUID TEMPERATURE. AFTER THIS TIME TAKE THE PICNOMETER OUT OF THE BATHROOM, DRY IT CAREFULLY, LEAVE IT ANOTHER FIFTEEN MINUTES AT ROOM TEMPERATURE AND WEIGH.

2.3.4 EMPTY THE PICNOMETER, RINSE WITH ACETONE AND DRY COMPLETELY WITH AIR CURRENT. LEAVE FIFTEEN MINUTES AT ROOM TEMPERATURE, COVER AND WEIGH.

AS DESCRIBED:

PA=XX_ENCODE_CASE_CAPS_LOCK_ON apparent weight of water=weight of the full picnometer-weight of the empty picnometer.

2.3.5 IN THE SAME WAY AS WITH WATER, PROCEEDS WITH THE LIQUID PROBLEM, BEING:

PW= WEIGHT OF PICNOMETER FILLED WITH LIQUID PROBLEM-WEIGHT OF THE EMPTY PICNOMETER.

2.4 CALCULATIONS

2.5 REMARKS

TO PREVENT THE SOLIDIFICATION OF THE SAMPLE, THE ROOM TEMPERATURE SHOULD BE CLOSE TO 25 DEGREES CELSIUS.

2.6 REFERENCES

1.

OFFICIAL METHODS OF ANALYSIS OF THE AOAC, 13. EDITION, 1984:

9,020-9.022.3. MELTING POINT OF NATURAL ESSENTIAL OIL (IN RAW MATERIAL) 3.1 PRINCIPLE

OBSERVATION, WITH A SUITABLE SYSTEM, OF THE TEMPERATURE AT WHICH THE ESSENTIAL OIL FUSION OCCURS, PREVIOUSLY FROZEN. AS THE ESSENTIAL OIL IS NOT A PURE COMPOUND, IT WILL NOT HAVE A FIXED MELTING POINT, BUT WILL MELT IN A GIVEN RANGE SINCE THE FIRST SYMPTOMS ARE OBSERVED IN THE CRYSTALS UNTIL THE SAMPLE BECOMES COMPLETELY LIQUID.

3.2 MATERIAL AND APARATOS

3.2.1 AGITATOR.

3.2.2 THERMOMETER OF 10 DEGREES CELSIUS-30 DEGREES CELSIUS, GRADUATED IN TENTHS OF A DEGREE.

3.2.3 GLASS CAPILLARY TUBES.

3.2.4 MAGNIFYING GLASS OR LENS.

3.2.5 1,000 MILLILITER BEAKER, LOW FORM.

3.3 PROCEDURE

THERE IS A SIMPLE MEASURING SYSTEM CONSISTING OF A MAGNETIC STIRRER WITH VARIABLE HEATING PLATE, A 600 MILLILITER BEAKER, A THERMOMETER AND A MAGNIFYING GLASS.

PREPARE A CLOSED CAPILLARY TUBE AT ITS END, WHERE A DROP OF THE ESSENTIAL OIL IS REACHED WITH THE HELP OF ANOTHER FINER CAPILLARY TUBE.

INTRODUCE THE TIP OF THE CAPILLARY TUBE INTO AN ICE BATH FOR TWO HOURS TO FREEZE.

FILL IN THE GLASS APPROXIMATELY HALF WITH DISTILLED WATER AND ENTER THE TIP OF THE CAPILLARY AND THE THERMOMETER SO THAT THEY ARE AT LEAST 30 MILLIMETERS BELOW THE SURFACE.

STARTING 8-10 DEGREES CELSIUS BELOW THE MELTING POINT OF THE SAMPLE, HEAT GENTLY TO INCREASE THE BATH TEMPERATURE 0.5 DEGREES CELSIUS/MINUTE, APPROXIMATELY, SIMULTANEOUSLY AGITATING THE BATH WATER WITH AID OF THE AGITATOR.

RECORD THE TEMPERATURE AT WHICH THE FULL MERGE OF THE SAMPLE OCCURS, WHICH WILL BE OBSERVED WITH THE HELP OF A MAGNIFYING GLASS OR MAGNIFYING LENS.

3.4 RESULT

THE MELTING POINT OF THE ESSENTIAL OIL WILL BE THE ARITHMETIC MEAN OF THREE SUCCESSIVE DETERMINATIONS WHICH MUST MATCH IN A RANGE OF 0.5 DEGREES CELSIUS.

3.5 REFERENCES

1.

OFFICIAL METHODS OF ANALYSIS OF THE AOAC, 14. EDITION, 1984:

28.014.4. ALCOHOLIC STRENGTH

4.1 PRINCIPLE

DISTILLATION OF THE PRODUCT AND SUBSEQUENT MEASURE OF THE DENSITY OF THE DISTILLATE BY AREOMETRY.

4.2 MATERIAL AND APARATOS

4.2.1 DISTILLATION APPARATUS AS IN THE OFFICIAL METHOD NUMBER 1.

4.2.2 ALKALHOMETER GRADUATED IN TENTHS OF D20 DEGREE, DULY CONTRASTED.

4.2.3 GRADUATED THERMOMETER IN TENTHS OF A DEGREE.

4.2.4 TRANSPARENT TEST TUBE OF 36 MILLIMETERS OF INTERNAL DIAMETER AND 320 MILLIMETERS OF HEIGHT.

4.2.5 THERMOSTATIC BATH AT 20 DEGREES CELSIUS.

4.3 PROCEDURE

4.3.1 DISTILLATE FROM DISTILLATE: BY OFFICIAL METHOD NUMBER 1.

4.3.2 AEROMETRIC DETERMINATION:

CLEAN AND DRY THE ALKALOMETER BEFORE USE.

POUR THE DISTILLED PROPERLY HOMOGENIZED INTO THE TEST TUBE.

INTRODUCE THE ALKALOMETER AND THE THERMOMETER INTO THE LIQUID.

WAIT TWENTY MINUTES FOR THE THERMOMETER, THE ALKALOMETER AND THE DISTILLATE TO ACQUIRE THE TEMPERATURE OF THE THERMOSTATIC BATH.

REACH THIS TEMPERATURE, REMOVE THE THERMOMETER, DRY THE ALKALOMETER STEM AND ALLOW IT TO BE INSERTED INTO THE DISTILLATE WITH A SLIGHT ROTATION MOVEMENT.

NOTE THE ALCOHOLIC STRENGTH READ.

REPEAT THESE OPERATIONS MULTIPLE TIMES. THE READINGS OBTAINED ARE VALID IF THEY HAVE AN APPROXIMATION OF 0.1 DEGREES ALCOHOLICS. IF NOT, PERFORM ADDITIONAL READINGS AND CALCULATE THE MEAN.

4.4 CALCULATIONS

THE ALCOHOLIC STRENGTH IS THE READING OBTAINED IN 4.3.2, EXPRESSED IN DEGREES AND TENTHS OF ALCOHOL DEGREE.

IF IN THE PRODUCTION OF THE DISTILLATE 100 MILLILITRES OF SAMPLE WERE DISTILLED TO 200 MILLILITRES, THE READING MUST BE MULTIPLIED BY TWO.

4.5 REFERENCES

1.

OFFICIAL METHODS OF ANALYSIS OF THE AOAC, 14. EDITION, 1984:

9.023-9.024.5. TOTAL SUGARS

5.1 PRINCIPLE

PRIOR DILUTION OF THE SAMPLE, INVERSION AND SUBSEQUENT ASSESSMENT BASED ON THE REDUCING ACTION OF THE SUGARS ON A CUPROALKALINE SOLUTION.

5.2 MATERIAL AND APARATOS

5.2.1 ERLENMEYER 250 MILLILITRES WITH FROSTED MOUTH.

5.2.2 USEFUL-LENGTH 30-CENTIMETER REFLUX REFRIGERANT.

5.2.3 REQUIRED MATERIAL FOR VOLUMETRY.

5.2.4 WATER BATH WITH THERMOSTAT.

5.3 REAGENTS

5.3.1 CONCENTRATED HYDROCHLORIC ACID 12 N.

5.3.2 SODIUM HYDROXIDE SOLUTION 12 N.

5.3.3 0.1 N. SODIUM HYDROXIDE SOLUTION

5.3.4 CUPROALCALIN SOLUTION:

DISSOLVE 25 GRAMS OF COPPER SULPHATE (II) PURE PENTAHYDRATE (SO4CU). 5H2O) IN 100 MILLILITRES OF DISTILLED WATER; 50 GRAMS OF CITRIC ACID MONOHYDRATE IN 300 MILLILITRES OF DISTILLED WATER AND 388 GRAMS OF SODIUM CARBONATE DECAHYDRATE BY 300-400 MILLILITRES OF HOT WATER.

CAREFULLY MIX THE SOLUTIONS OF CITRIC ACID AND SODIUM CARBONATE. IMMEDIATELY ADD THE COPPER SULPHATE SOLUTION AND LEAD TO A LITRE.

5.3.5 POTASSIUM IODIDE SOLUTION AT 30 PER 100 (P/V).

STORE IN BOTTLE TOPAZ. IT IS ADVISABLE NOT TO PREPARE MORE THAN YOU WILL CONSUME DAILY.

5.3.6 SULFURIC ACID SOLUTION AT 25 PER 100 BY VOLUME.

5.3.7 STARCH 5 GRAM/LITRE. TO PREPARE THIS SOLUTION, KEEP IT ON THE BOIL FOR TEN MINUTES. ADD 200 GRAMS OF SODIUM CHLORIDE PER LITRE TO ENSURE ITS PRESERVATION.

5.3.8 0.1 N. SODIUM THIOSULFATE SOLUTION

5.4 PROCEDURE

PIPETEAR 10 MILLILITRES OF ANISE (OR THE APPROPRIATE VOLUME DEPENDING ON THE SUGAR CONTENT) IN A 500 MILLILITRE FLASK, BEING MADE UP WITH DISTILLED WATER (SOLUTION A).

IN A FLASK OF 200 MILLILITRES, 25 MILLILITRES OF THE SOLUTION ARE PIPETED WITH DISTILLED WATER (SOLUTION B).

INVESTING IS DONE AS FOLLOWS:

IN A FLASK OF 50 MILLILITRES PIPETTE 20 MILLILITRES OF SOLUTION B. 3 MILLILITRES OF CONCENTRATED HYDROCHLORIC ACID ARE ADDED, TAKING FOR FIVE MINUTES A BATH AT 75 DEGREES CELSIUS.

GETS TAKEN OUT AND LEFT TO COOL.

PHENOLPHTHALEIN IS ADDED AND NEUTRALIZED WITH SODIUM HYDROXIDE SOLUTION 12 N UP TO NEAR THE POINT OF VIRAGE AND SUBSEQUENTLY WITH SODIUM HYDROXIDE SOLUTION 0.1 N TO VIRAL. IT IS ENRASE WITH DISTILLED WATER AND IS DERIVED FROM THE ANALYSIS OF TOTAL REDUCING SUGARS (SOLUTION M).

IN AN ERLENMEYER 25 MILLILITRES OF THE SOLUTION CUPROALKALINE AND 25 MILLILITRES OF THE SOLUTION M ARE PUT TO THE BOIL WITH REFLUX REFRIGERANT KEEPING THE BOIL TEN MINUTES; IT COOLS QUICKLY, ADD 2 MILLILITRES OF STARCH, 10 MILLILITRES OF POTASSIUM IODIDE SOLUTION AND 25 MILLILITRES OF SULPHURIC ACID AT 25 PER 100, IMMEDIATELY BEING MEASURED WITH SODIUM THIOSULFATE SOLUTION 0.1 N. THE TITRATION WITH A TARGET IS REPEATED, REPLACING THE 25 MILLILITRES OF SOLUTION M FOR 25 MILLILITRES OF DISTILLED WATER.

5.5 CALCULATIONS

BE D THE DIFFERENCE BETWEEN WHAT WAS SPENT BY THE TARGET AND WHAT WAS SPENT BY THE SAMPLE.

THE CONCENTRATION OF TOTAL SUGARS IN THE SAMPLE IS OBTAINED FROM THE VALUE OF D IN THE TABLE, INTERPOLATING IF NECESSARY AND MULTIPLYING THE VALUE OBTAINED BY 40 (DILUTION FACTOR).

CONCENTRATION TOTAL SUGARS GRAMS/LITRES VALUE OBTAINED IN TABLE 40.5.6 OBSERVATIONS

5.6.1 IN THE CASE OF TAKING A VOLUME OTHER THAN ANISE, TAKE INTO ACCOUNT FOR CALCULATIONS THE DILUTION FACTOR.

5.7 REFERENCES

1.

RECUEIL DES METHODS INTERNATIONALES D' ANALYSE DES VINS OIV A4-126-127.

6. ANETOL

6.1 PRINCIPLE

SEPARATION, IDENTIFICATION AND QUANTIFICATION OF ANETOL BY GAS CHROMATOGRAPHY.

6.2 MATERIAL AND APARATOS

6.2.1 GAS CHROMATOGRAPH EQUIPPED WITH FLAME IONIZATION DETECTOR.

6.2.2 STAINLESS STEEL COLUMN 3 METERS IN LENGTH 1/8 INCHES INTERNAL DIAMETER, FILLED FROM 52 TO 4 PER 100 OVER CHROMOSOB. AW. DMA, 100-120 MALLAS.

6.2.3 MICROSYRINGE OF 1

6.3 REAGENTS

6.3.1 TRANS-ANETOL PATTERN WITH A CIS-ANETOL CONTENT OF LESS THAN 0.5 PER 100.

6.3.2 MENTOL.

6.3.3 ABSOLUTE ETHANOL.

6.4 PROCEDURE

6.4.1 CALIBRATION CURVE.

6.4.1.1 INTERNAL PATTERN.

WEIGH 0.25 GRAMS OF MENTHOL AND LEAD TO A 100 MILLILITRE GRADUATED FLASK, DISSOLVE IN 40 MILLILITRES OF ABSOLUTE ETHANOL AND MAKE UP WITH DISTILLED WATER.

6.4.1.2 PATTERN SOLUTION.

WEIGH 1 GRAM OF ANETOL AND DISSOLVE IN 400 MILLILITRES OF ABSOLUTE ETHANOL, MAKE UP TO 1 LITRE WITH DISTILLED WATER. THIS PATTERN MUST BE AT THE TIME OF ITS NEWLY PREPARED UTILIZATION.

6.4.1.3 CALIBRATED SOLUTION. FROM THE SOLUTION 6.4.1.2 PREPARE SOLUTIONS CONTAINING 0.1, 0.25, 0.50, 0.75 AND 1 GRAM/LITRE IN 40 DEGREE ALCOHOL.

TO 100 MILLILITRES OF THESE SOLUTIONS IS ADDED 5 MILLILITRES OF THE SOLUTION 6.4.1.1.

THESE SOLUTIONS ARE INJECTED INTO THE CHROMATOGRAPH BY OBTAINING THE CALIBRATION CURVE, REPRESENTING:

PATTERN AREA VERSUS CONCENTRATIONS 6.4.2 APPROXIMATE CHROMATOGRAPHIC CONDITIONS.

OVEN TEMPERATURE: 130 DEGREES CELSIUS.

INJECTOR TEMPERATURE: 220 DEGREES CELSIUS.

DETECTOR TEMPERATURE: 220 DEGREES CELSIUS.

6.4.3 SAMPLE PREPARATION.

TAKE 100 MILLILITRES OF THE DISTILLATE, ACCORDING TO METHOD 1, ADD 5 MILLILITRES OF INTERNAL STANDARD AND INJECT INTO THE CHROMATOGRAPH.

6.5 CALCULATIONS

6.5.1 CALCULATE THE RATIO AND LEAD TO THE CALIBRATION CURVE, OBTAINING THE VALUE OF THE ANETHOL CONCENTRATION IN THE TEST SAMPLE.

6.5.2 A FIXED CONCENTRATION PATTERN OF ANETOL CAN ALSO BE USED, WITH THE RESPONSE FACTOR AND SAMPLE CONCENTRATION BEING CALCULATED BY APPLYING THE FOLLOWING FORMULAE:

CALCULATION OF THE SAMPLE ' S ANETOL CONCENTRATION SHOULD BE PERFORMED WITH AN APPROXIMATE TO THE EXPECTED PATTERN OF WEALTH. IF THIS IS NOT KNOWN, A PRE-INJECTION SHOULD BE MADE TO FIX THE LEVEL OF ANETHOL BY REFERRING TO THE 1 GRAM/LITRE PATTERN AND ONCE THE STANDARD IS EVALUATED, IF

NEED, OF CONCENTRATION CLOSE TO THE SAMPLE, OBTAINING THE FINAL RESULT BY REFERENCE TO THIS SECOND PATTERN.

6.6 REFERENCES

1. JOSE RAMON GARCIA IRON AND GERTRUDIS MARIN ESTEBAN. REPORT OF THE ARBITRAL LABORATORY OF THE SUB-DIRECTORATE-GENERAL OF AGRICULTURAL LABORATORIES.

7. METHANOL

7.1 PRINCIPLE

SEPARATION, IDENTIFICATION AND QUANTIFICATION OF METHANOL BY GASEOUS CHROMATOGRAPHY.

7.2 MATERIAL AND APARATOS

7.2.1 GAS CHROMATOGRAPH EQUIPPED WITH FLAME IONIZATION DETECTOR.

7.2.2 CHROMATOGRAPHIC COLUMN.

COLUMN OF 4 METERS IN LENGTH, 1/8 INCHES OF INTERNAL DIAMETER, FILLED WITH CARBOWAX 1500, AT 15 PER 100 ON CHROMOSORLO W, 80-100 MESHES, WASHED TO ACIDS.

7.2.3 REACTIVE 17.3 MICROSYRINGES

7.3.1 METHANOL CHROMATOGRAPHIC PURITY.

7.3.2 4-METHYL-2-PENTANOL.

7.3.3 CHROMATOGRAPHIC PURITY ETHANOL.

7.4 PROCEDURE

7.4.1 DISTILLED FROM DISTILLATE AS IN OFFICIAL METHOD NUMBER 1.

7.4.2 CHROMATOGRAPHIC CONDITIONS:

THE ORIENTATION CHROMATOGRAPHIC CONDITIONS ARE: CARRIER GAS; NITROGEN, 20 MILLIITROS/MIN, OVEN TEMPERATURE, 85 DEGREES CELSIUS; INJECTOR AND DETECTOR TEMPERATURE, 150 DEGREES CELSIUS.

7.4.3 CALIBRATED:

7.4.3.1 INTERNAL PATTERN: 5 GRAMOS/LITRE SOLUTION OF 4-METHYL-2-PENTANOL IN 40 DEGREE ETHANOL.

7.4.3.2 PATTERN SOLUTION: PREPARE A METHANOL SOLUTION IN ETHANOL AT 40 DEGREES, OF SIMILAR CONCENTRATION AS EXPECTED IN THE SAMPLE. 10 MILLILITRES OF THIS SOLUTION ADD 1 MILLILITRE OF THE INTERNAL STANDARD SOLUTION 7.4.3.1 AND INJECT 1 1 IN THE CHROMATOGRAPH.

7.4.4 SAMPLE PROBLEM: 10 MILLILITRES OF THE ANISE DISTILLATE, CARRIED TO 40 DEGREES, IS ADDED 1 MILLILITRE OF INTERNAL STANDARD 7.4.3.1 AND INJECT 1 1 IN THE SAME CONDITIONS AS IT WAS DONE FOR THE CALIBRATION.

7.6 REFERENCES

1. , 14. EDITION, 1984:

5,008-5,011.

8. NITROGENOUS BASES

8.1 PRINCIPLE

FIXING OF NITROGEN AND AMMONIA BASES BY PHOSPHORIC ACID IN THE DISTILLATE OF THE SAMPLE.

RELEASE OF THESE BY SOLUTION OF SODIUM HYDROXIDE AND ASSESSMENT WITH HYDROCHLORIC ACID.

THE DETECTION LIMIT FOR THIS METHOD IS 1 MILLIGRAM/LITRE.

8.2 MATERIAL AND APARATOS

8.2.1 SIMPLE DISTILLER WITH DEVICES FOR ADDITION (FIGURE 8.1).

8.2.2 TEST TUBE OF 100 MILLILITRES OF CAPACITY.

8.2.3 GRINDING MOUTH TEST TUBES OF 30 MILLILITRES OF CAPACITY.

8.2.4 25-MILLILITER BURETA.

8.2.5 PIPETTES OF 50 MILLILITRES OF CAPACITY.

8.2.6 FLASK OF 50 MILLILITRES.

8.3 REAGENTS

8.3.1 PHOSPHORIC ACID (D 1,34 GRAMS/MILLILITRE).

8.3.2 DISSOLUTION OF SODIUM HYDROXIDE TO 1 PER 100.

8.3.3 DISSOLUTION OF HYDROCHLORIC ACID 0.01 N.

8.3.4 DISSOLUTION OF METHYL RED: WEIGH 0.1 GRAMS OF METHYL RED, DISSOLVE IN 50 MILLILITRES OF ETHANOL OF 90 PER 100 V/V).

8.3.5 BIDESTIZED AND DEIONIZED WATER PH 6.5-7.5.

8.4 PROCEDURE

OBTAIN THE DISTILLATE OF 100 MILLILITRES OF SAMPLE, OPERATING ACCORDING TO METHOD OF ANALYSIS NUMBER 1.

IN A ROUND-BOTTOM FLASK OF 250 MILLILITRES ENTER 60 MILLILITRES OF THE DISTILLATE, 40 MILLILITRES OF DISTILLED WATER AND TWO DROPS OF PHOSPHORIC ACID; DISTILL, COLLECTING THE FIRST 80 MILLILITRES OF THE DISTILLATE IN 8.2.2 AND Discard them.

COOL THE FLASK AND THE COLD RESIDUE ADD 20 MILLILITRES OF SODIUM HYDROXIDE DISSOLUTION TO 1 PER 100. DISTILLING AGAIN, COLLECTING ABOUT 10 MILLILITRES OF DISTILLATE IN A TEST TUBE, IN WHICH TWO MILLILITRES OF DISTILLED WATER AND ONE DROP OF METHYL RED DISSOLUTION HAVE BEEN PREVIOUSLY INTRODUCED, WITH THE DISTILLATE FALLING INTO THE BOTTOM OF THE TUBE WITH THE AID OF AN ELONGATED.

THEN ASSESS WITH DISSOLUTION OF HYDROCHLORIC ACID 0.01 N UNTIL THE INDICATOR TURN.

PERFORM THE SAME PROCEDURE ON AN ETHANOL TARGET AT 40 PER 100.

8.6 REFERENCES

1. , 1978, PAGE 17.

9. ESSENTIAL OILS

9.1 PRINCIPLE

EXTRACTION OF ESSENTIAL OILS AND QUANTIFICATION.

9.2 MATERIAL AND APARATOS

9.2.1 500 MILLILITER DECANTING FUNNEL.

9.2.2 DESICCATOR WITH DEVICE FOR MAKING VACUUM OR ROTAVAPOR.

9.2.3 100 MILLILITRE HEART-SHAPED FLASKS WITH PROPER MOUTH TO ADAPT TO ROTAAVAPOR.

9.3 REAGENTS

9.3.1 N-PENTH.

9.3.2 SODIUM CHLORIDE.

9.3.3 ETHYL ETHER.

9.3.4 ANETOL PATTERN.

9.4 PROCEDURE

TAKE 40 MILLILITRES OF ANISE AND INTRODUCE INTO A DECANTATION FUNNEL ADDING 160 MILLILITRES OF DISTILLED WATER, 40 GRAMS OF CLNA AND 60 MILLILITRES OF N-PENTANE. SHAKE AND LET DECANTAR FOR ABOUT TEN MINUTES.

ONCE THE PHASES ARE SEPARATED, TAKE THE CORRESPONDING ONE TO THE PENTANE. ADD SODIUM SULFATE ANHYDROUS. FILTER. TAKE THE ROTAVAPOR TO EVAPORATE THE PENTANE, WITHOUT THE BATH TEMPERATURE REACHING 30 DEGREES CELSIUS.

DISSOLVE THE RESIDUE OBTAINED IN 5 MILLILITRES OF ETHYL ETHER AND INJECT INTO THE CHROMATOGRAPH UNDER THE SAME CONDITIONS OF METHOD NUMBER 6. NORMALIZE THIS INJECTION OBTAINED THE PERCENTAGE OF ANETOL.

9.6 REFERENCES

1. . A.

MAUREL. ANNALES DES FALSIFICATIONS DE L' EXPERTISE CHIMIQUE ET TOXICOLOGUIQUE, 1941.

2. INTERNAL REPORT OF THE WORKING GROUP OF OFFICIAL METHODS OF ANALYSIS OF THE MINISTRY OF AGRICULTURE, FISHERIES AND FOOD.

10. HYDROXYMETHYLFURFURAL AND FURFURAL

10.1 PRINCIPLE

SEPARATION AND QUANTIFICATION BY LIQUID-LIQUID CHROMATOGRAPHY OF HYDROXYMETHYLFURFURAL AND FURFURAL.

10.2 MATERIAL AND APARATOS

10.2.1 LIQUID-LIQUID CHROMATOGRAPH WITH DETECTOR U. V.

10.2.2 SHERISORB COLUMN SDGS 5 OF 20 0.46 CENTIMETERS OR SIMILAR COLUMN OF REVERSE PHASE.

10.2.3 MICROSYRINGE OF 25 OR 50 1 OR ANY SUITABLE INJECTION SYSTEM ACCORDING TO LOOP AND INJECTOR.

10.2.4 SOLVENT FILTRATION EQUIPMENT FOR LIQUID-LIQUID CHROMATOGRAPHY WITH A FILTER OF 0.45 M OR SIMILAR.

10.2.5 SAMPLE FILTRATION EQUIPMENT WITH 0.5 M OR SIMILAR FILTERS.

10.3 REAGENTS

10.3.1 METHANOL GRADE HPLC.

10.3.2 MONOPOTASSIUM PHOSPHATE BUFFER AND DIPOTASSIUM PHOSPHATE.

DISSOLVE 1.25 GRAMS OF KH2PO4 AND 1.25 GRAMS OF K2HPO4 IN WATER UP TO ONE LITER.

10.3.3 HYDROXYMETHYLFURFURAL PATTERN AND RECENTLY DISTILLED FURFURAL.

10.4 PROCEDURE

10.4.1 GUIDANCE CHROMATOGRAPHIC CONDITIONS:

MOBILE PHASE. METHANOL SOLUTION AT 10 PER 100 (V/V) IN BUFFER SOLUTION (10.3.2) FLOW: 0.8 MILILITROS/MN.

WAVELENGTH: 285 NM.

APPROXIMATE RETENTION TIME: SEVEN MINUTES FOR HYDROXYMETHYLFURFURAL AND TEN MINUTES FOR FURFURAL.

10.4.2 STANDARD SOLUTIONS:

PREPARATION HYDROXYMETHYLFURFURAL AND FURFURAL SOLUTIONS IN METHANOL OF APPROXIMATE CONCENTRATIONS AT 5 MILLIGRAM/LITRE.

10.4.3 DILUTE 5 MILLILITRES OF THE SAMPLE IN METHANOL TO 25 MILLILITRES.

FILTER AND INJECT DIRECTLY.

10.5.

CALCULATIONS

CALCULATE THE CONCENTRATION OF HYDROXYMETHYLFURFURAL AND FURFURAL EXPRESSED IN MILLIGRAM/LITRE USING THE FOLLOWING FORMULAE.

ARSENIC

11.1 PRINCIPLE

THE SAMPLE IS SUBJECTED TO ACID DYSMANAGEMENT WITH A MIXTURE OF NITRIC AND SULPHURIC ACID.

ARSENIC DETERMINATION IS PERFORMED BY ATOMIC ABSORPTION SPECTROPHOTOMETRY, WITH HYDRIDE GENERATOR.

11.2 MATERIAL AND APARATOS

11.2.1 BALANCES WITH ACCURACY OF 0.001 GRAMS AND 0.1 GRAMS.

11.2.2 VOLUMETRIC 50 AND 100 MILLILITRES OF CAPACITY.

11.2.3 DOUBLE-CAPACITY PIPETTES.

11.2.4 MATRACES OF 250 MILLILITRES.

11.2.5 ATOMIC ABSORPTION SPECTROPHOTOMETER EQUIPPED WITH HYDRIDE GENERATOR SYSTEM.

11.2.6 ELECTROLESS DISCHARGE LAMP.

11.2.7 UNELECTRODES DISCHARGE SOURCE.

11.2.8 GRAPHIC LOGGER.

11.3 REAGENTS

ONLY PURITY GRADE REAGENTS ARE USED FOR ANALYSIS AND DISTILLED WATER.

11.3.1 HYDROCHLORIC ACID (D 1,19 GRAMS/MILLILITER).

11.3.2 DISSOLUTION OF HYDROCHLORIC ACID.

DISSOLVE 32 MILLILITRES OF HYDROCHLORIC ACID (D 1.19 GRAMOS/MILLI TRO), WITH DISTILLED WATER UP TO A VOLUME OF 100 MILLILITRES.

11.3.3 DISSOLUTION OF HYDROCHLORIC ACID.

DISSOLVE 15 MILLILITRES OF HYDROCHLORIC ACID (D 1,19 GRAMS/MILLILITRE) WITH DISTILLED WATER UP TO A VOLUME OF 1,000 MILLILITRES.

11.3.4 NITRIC ACID (D 1.40 GRAMS/MILLILITRE).

11.3.5 SULPHURIC ACID (D 1,84 GRAMS/MILLILITRE).

11.3.6 DISSOLUTION OF SODIUM HYDROXIDE TO 1 PER 100.

WEIGH 1 GRAM OF SODIUM HYDROXIDE AND DISSOLVE IT WITH DISTILLED WATER UP TO A VOLUME OF 100 MILLILITRES.

11.3.7 SODIUM BOROHYDRIDE DISSOLUTION AT 3 PER 100.

WEIGH THREE GRAMS OF SODIUM BOROHYDRIDE AND DISSOLVE IT UP TO 100 MILLILITRES WITH SODIUM HYDROXIDE AT 1 PER 100.

11.3.8 DISSOLUTION TO 1 PER 100 OF DISODIUM ETHYLENDINITRILETETRAACETATE, DIHYDRATE (TRITIPLEX III).

WEIGH A GRAM OF TRITIPLEX III AND DISSOLVE IT UP TO 100 MILLILITRES WITH DISTILLED WATER.

11.3.9 DISSOLUTION OF POTASSIUM HYDROXIMUM TO 20 PER 100.

WEIGH 20 GRAMS OF POTASSIUM HYDROXIDE AND DISSOLVE IT WITH DISTILLED WATER UP TO A VOLUME OF 100 MILLILITRES.

11.3.10 DISSOLUTION OF SULPHURIC ACID AT 20 PER 100 (V/V).

DILUTE 20 MILLILITRES OF SULPHURIC ACID (11.3.5) WITH DISTILLED WATER UP TO A VOLUME OF 100 MILLILITRES.

11.3.11 DISSOLUTION OF SULPHURIC ACID AT 1 PER 100 (V/V).

DILUTE 1 MILLILITRE OF SULPHURIC ACID (11.3.5) WITH DISTILLED WATER UP TO A VOLUME OF 100 MILLILITRES.

11.3.12 CONCENTRATION ARSENIC PATTERN 1 GRAMO/LITRO.

DISSOLVE 0.132 GRAMS OF ARSENIC TRIOXIDE OF 2.5 MILLILITRES OF POTASSIUM HYDROXIDE AT 20 PER 100 (11.3.9), NEUTRALISE WITH SULPHURIC ACID AT 20 PER 100 (11.3.10), DILUTE UP TO 100 MILLILITRES WITH SULPHURIC ACID 1 PER 100 (11.3.11).

11.3.13 CONCENTRATION ARSENIC PATTERN 10 MILLIGRAMS/LITRES.

PIPETEAR 1 MILLILITRE OF THE ARSENIC STANDARD SOLUTION (11.3.12) IN A 100 MILLILITRE GRADUATED FLASK. DILUTE TO THE ROOT WITH DISTILLED WATER.

11.3.14 CONCENTRATION ARSENIC PATTERN 0.1 MILLIGRAMS/LITRES.

PIPETEAR 1 MILLILITRE OF THE ARSENIC SOLUTION (11.3.13) IN A 100 MILLILITRE GRADUATED FLASK. DILUTE TO THE ROOT WITH WATER DISTILLING DA.11.4 PROCEDURE

11.4.1 PREPARATION OF THE SAMPLE.

IN A MATRAZA OF 250 MILLILITERS INTRODUCE 4 GRAMS OF SAMPLE WITH 20 MILLILITRES OF NITRIC ACID (11.3.4) AND 5 MILLILITRES OF SULPHURIC ACID (11.3.5).

BRING THE BOIL TO REDUCE THE VOLUME TO 5 MILLILITRES.

ALLOW TO COOL AND DISSOLVE WITH DISTILLED WATER IN A 50 MILLILITRE FLASK THE RESULTING SOLUTION.

11.4.2 WHITE PREPARATION AND WORKING PATTERNS.

IN A MATRAZ , INTRODUCE 5 MILLILITRES OF THE ARSENIC SOLUTION (11.3.14) AND SUBMIT IT TO THE SAME TREATMENT AS THE SAMPLE.

1 MILLILITRE OF THE SOLUTION CONTAINS TEN NANOGRAMS OF ARSENIC.

PREPARE A TARGET WITH ALL THE REAGENTS USED FOLLOWING THE TREATMENT GIVEN TO THE SAMPLE.

11.4.3 CONDITIONS OF THE SPECTROPHOTOMETER.

TURN ON THE POWER SUPPLY OF THE NON-ELECTRODE DISCHARGE LAMPS IN SUFFICIENT TIME TO STABILIZE THE POWER OF THE LAMP.

TURN ON THE SPECTROPHOTOMETER, ADJUST THE WAVELENGTH OF 193.7 NM, PLACING THE GRID ACCORDING TO THE CONDITIONS OF THE APPARATUS.

TURN ON THE HYDRIDE GENERATOR, PLACING THE CELL TEMPERATURE AT 900 C, WAITING UNTIL THAT TEMPERATURE IS REACHED.

THE CONDITIONS OF THE HYDRIDE GENERATOR ARE ADJUSTED ACCORDING TO THE SPECIFICATIONS OF THE APPARATUS.

ADJUST THE ARGON FLOW ACCORDING TO THE CHARACTERISTICS OF THE APPARATUS.

TURN ON THE LOGGER.

11.4.4 DETERMINATION.

ARSENIC CONCENTRATION DETERMINATIONS ARE MADE BY THE METHOD OF ADDITION OF PATTERNS, BY MEANS OF DUPLICATE MEASURES IN THE SPECTROPHOTOMETER UNDER THE CONDITIONS SPECIFIED IN (11.4.3) BY ADDING THE REACTION FLASK 3 MILLILITRES OF SOLUTION (11.4.1) PLUS 3 MILLILITRES OF THE SOLUTION (11.3.2) AND 10 MILLILITRES OF THE SOLUTION (11.3.8). AS INTERNAL PATTERNS ARE USED 10, 20 AND 50 NG OF AS.

WASH FLASKS BEFORE AND AFTER EACH USE WITH HYDROCHLORIC ACID (11.3.3).

IN CONSTRUCTING THE ADDITION CHART, YOU MUST DISCOUNT THE ABSORBANCE VALUE OF THE TARGET OBTAINED UNDER THE SAME CONDITIONS ABOVE, BUT ADD 3 MILLILITRES OF THE WHITE SOLUTION.

12. Lead

12.1 PRINCIPLE

DETERMINATION OF LEAD BY ATOMIC ABSORPTION.

12.2 MATERIAL And APARATOS

12.2.1 ATOMIC ABSORPTION SPECTROPHOTOMETER.

12.2.2 LEAD LAMP.

12.2.3 PLATINUM, QUARTZ OR SIMILAR CAPSULES (150 MILLILITRES OF CAPACITY, APPROXIMATELY).

12.2.4 SAND BATH OR HEATING PLATE WITH TEMPERATURE REGULATION OR STOVE WITH TEMPERATURE CONTROL IN THE RANGE OF 50 TO 150 C.

12.2.5 MUFLA. IT IS SUITABLE TO HAVE IT INSIDE THE SMOKE EXTRACTION WINDOW.

12.2.6 FLASKS OF 10, 100 AND 1,000 MILLILITRES OF CAPACITY.

12.3 REAGENTS

12.3.1 SULPHURIC ACID OF 96 PER 100 (D 1,835) GRAMS/MILLILITRE.

12.3.2 NITRIC ACID 70 PER 100 (D 1,413) GRAMS/MILLILITRE.

12.3.3 NITRIC ACID AT 1 PER 100 IN DISTILLED WATER (V/V).

12.3.4 STANDARD SOLUTION OF 1,000 MILLIGRAMS OF PB/LITRE. DISSOLVE 1,598 GRAMS OF LEAD NITRATE (II) BY MAKING UP TO 1,000 MILLILITRES WITH NITRICOAL ACID 1 PER 100.

12.4 PROCEDURE

12.4.1 PREPARATION OF THE SAMPLE. WEIGH 20 GRAMS OF THE SAMPLE IN THE CAPSULE (12.2.3); BRING IT TO EVAPORATION TO SIRUPOSE CONSISTENCY IN A SAND BATH (12.2.4). THEN ADD 2 MILLILITRES OF SULPHURIC ACID AND SLOWLY CARBONISE THE RESIDUE IN THE SAND OR PLATE BATH. THEN INSERT THE CAPSULE INTO THE MUFLA AND KEEP IT FOR TWO HOURS AT 450 C; AFTER THAT TIME, REMOVE IT AND LET IT COOL. ADD 1 MILLILITRE OF DISTILLED WATER, EVAPORATE IN THE SAND BATH OR PLATE AND INSERT INTO THE MUFLA AND KEEP IT FOR TWO HOURS AT 450 C; AFTER THAT TIME, REMOVE IT AND LET IT COOL. ADD 1 MILLILITRE OF DISTILLED WATER, EVAPORATE IN THE SAND BATH OR PLATE AND INTRODUCE INTO THE MUFLA, REPEATING THIS OPERATION UNTIL OBTAINING WHITE ASHES. A SLIGHT RED MARRON-REDDISH IN THE ASHES (POSSIBLY FE2O3 IS ACCEPTED AND DOES NOT REQUIRE FURTHER TREATMENT.

THEN DISSOLVE THE ASHES WITH 1 MILLILITRE OF CONCENTRATED NITRIC ACID AND 2 MILLILITRES OF DISTILLED WATER. BRING THE SOLUTION TO A 10 MILLILITER FLASK, WASH THE CAPSULE WITH DISTILLED WATER AND ADD THE WASH WATER TO THE ROOT, SUBSEQUENTLY FILTERING.

12.4.2 PATTERN CURVE CONSTRUCTION. DILUTE APPROPRIATE ALIQUOTS OF THE STANDARD SOLUTION (12.3.4) WITH NITRIC ACID TO 1 PER 100 (V/V) TO OBTAIN A CONCENTRATION CURVE 2, 4, 6, 8 AND 10 MILLIGRAM/LITRES.

12.4.3 DETERMINATION. OPERATE ACCORDING TO THE SPECIFICATIONS OF THE APPARATUS, USING AIR-ACETYLENE FLAME. MEASURE THE ABSORBANCES OF THE SAMPLE AND PATTERNS AT 283 NM. IF THE SOLUTION IS VERY CONCENTRATED, DILUTE IT WITH NITRIC ACID AT 1 PER 100.

12.5 CALCULATIONS

CALCULATE THE LEAD CONTENT, EXPRESSED IN MILLIGRAM/LITRE BY COMPARISON WITH THE CORRESPONDING STANDARD CURVE AND TAKING INTO ACCOUNT THE CONCENTRATION OR DILUTION FACTOR.

12.6 REFERENCES

1.

OFFICIAL WINE ANALYSIS METHODS. MINISTRY OF AGRICULTURE, PAGE 134 (I), 1976.

13. CNC

13.1 PRINCIPLE

DETERMINATION OF ZINC BY ATOMIC ABSORPTION AFTER SAMPLE MINERALIZATION.

13.2 MATERIAL And APARATOS

13.2.1 ATOMIC ABSORPTION SPECTROPHOTOMETER.

13.2.2 LAMP OF CCN.

13.2.3 USED FOR PB IN (12.2.3), (12.2.4), (12.2.5) AND (12.2.6) .13.3 REAGENTS

13.3.1 USED FOR LEAD IN (12.3.1), (12.3.2) AND (12.3.3).

13.3.2 STANDARD SOLUTION OF 1,000 MILLIGRAMS OF CINC/LITRE. DISSOLVE 1,000 GRAMS OF CINRUSH IN THE MINIMUM REQUIRED VOLUME OF NITRIC ACID (1: 1) AND DILUTE TO 1 LITRE WITH NITRIC ACID 1 PER 100 (V/V).

13.4 PROCEDURE

13.4.1 PREPARATION OF THE SAMPLE. AS IN (12.4.1).

13.4.2 PATTERN CURVE CONSTRUCTION. DILUTE ALIQUOT PARTS OF THE STANDARD SOLUTION (13.3.2) WITH NITRIC ACID TO 1 PER 100 FOR SOLUTIONS OF 0.5, 1, 1.5 AND 2 MILLIGRAM/LITRE.

13.4.3 DETERMINATION. SAME AS FOR LEAD. THE READING WILL BE DONE AT 213.8 NM.

13.5 CALCULATIONS

STARTING FROM THE ABSORBENCY VALUES OBTAINED TO FIND ZINC CONCENTRATIONS FOR THE SAMPLE, TAKING INTO ACCOUNT THE CONCENTRATION OR DILUTION FACTOR.

13.6 REFERENCES

1.

H. E. PARKER. (1963), 13.

14. COBRE

14.1 PRINCIPLE

DETERMINATION OF COPPER BY ATOMIC ABSORPTION AFTER SAMPLE MINERALIZATION.

14.2 MATERIAL AND APARATOS

14.2.1 ATOMIC ABSORPTION SPECTROPHOTOMETER.

14.2.2 COPPER LAMP.

14.2.3 USED FOR LEAD IN (12.2.3), (12.2.4), (12.2.5) AND (12.2.6).

14.3 REAGENTS

14.3.1 USED FOR LEAD IN (12.3.1), (12.3.2) AND (12.3.3).

14.3.2 STANDARD SOLUTION OF 1,000 MILLIGRAMS OF COBRE/LITRE. DISSOLVE 1,000 GRAMS OF PURE COPPER IN THE MINIMUM VOLUME REQUIRED NO3 (1: 1) AND DILUTE TO 1 LITRE WITH NITRIC ACID 1 PER 100 (V/V).

14.4 PROCEDURE

14.4.1 PREPARATION OF THE SAMPLE. AS IN (12.4.1).

14.4.2 PATTERN CURVE CONSTRUCTION. DILUTE ALIQUOT PARTS OF THE STANDARD SOLUTION (14.3.2) WITH NITRIC ACID OF 1 PER 100 TO OBTAIN SOLUTIONS CONTAINING 1 TO 5 MILLIGRAMS OF COBE/LITRE.

14.4.3 DETERMINATION. SAME AS FOR LEAD. MEDIR AT 324.7 NM.

14.5 CALCULATIONS

STARTING FROM THE ABSORBANCE VALUES OBTAINED FOR THE SAMPLE, FIND BY THE PATTERN CURVE THE COPPER CONCENTRATIONS OF THE SAMPLE.

14.6 REFERENCES

1.

H. E. PARKER: (1963). 13.

2.

F. ROUSSELET: . ED.

PARIS (1968), PAGINAS 59-144.