BY ORDER OF THE MINISTRY OF AGRICULTURE ON 1 AUGUST 1979 ("OFFICIAL STATE BULLETIN" OF 28) THE REGULATION OF CIDERS AND OTHER APPLE-DERIVED BEVERAGES WAS APPROVED, IN ACCORDANCE WITH ARTICLE 22 OF THE MINISTRY OF AGRICULTURE TO ESTABLISH THE ANALYTICAL METHODS TO BE USED IN THE VERIFICATION OF THE PRODUCTS COVERED BY THE SAID ORDER.

CONSEQUENTLY AND AFTER THE APPROPRIATE STUDIES CARRIED OUT BY THE RELEVANT WORKING GROUP OF THE COORDINATING COMMISSION OF LABORATORIES AND METHODS OF ANALYSIS,

THIS MINISTRY HAS PROVIDED THE FOLLOWING:

ARTICLE 1. THE METHODS OF ANALYSIS OF CIDERS AND OTHER APPLE-DERIVED BEVERAGES LISTED IN ANNEX I ARE HEREBY APPROVED AS OFFICIAL.

ART. 2. ALL PROVISIONS OF EQUAL OR LOWER RANK TO THAT OF THIS MINISTERIAL ORDER SHALL BE REPEALED AS SOON AS THEY OBJECT TO IT.

MADRID, 30 JUNE 1980. -LAMO DE ESPINOX.

ANNEX 1

1. REMOVAL OF CARBON DIOXIDE.

1.1. PRINCIPLE:

DEGASSING OF CIDER BY AGITATION AND FILTRATION.

1.2. MATERIAL AND APARATOS:

1.2.1. MATRAZ ERLENMEYER.

1.2.2. FILTER PAPER.

1.2.3. EMBUDO.

1.3. PROCEDURE:

THE VOLUME OF CIDER TO BE USED, PREVIOUSLY TAKEN AT 17-20 GRAD. C, VERTERLO IN A DOUBLE-CAPACITY ERLENMEYER FLASK OF THE VOLUME USED. SHAKE FOR TEN MINUTES AND THEN FILTER THROUGH A FUNNEL COVERED IN DRY FILTER PAPER.

2. ACTUAL ALCOHOLIC STRENGTH.

REMOVE CARBON DIOXIDE AS INDICATED IN METHOD NUMBER 1 AND PROCEED AS INDICATED IN THE OFFICIAL WINE METHODS NUMBERS 5 (A) (OFFICIAL STATE BULLETIN OF 22 JULY 1977) AND 5 (B) (OFFICIAL STATE BULLETIN) OF 23 JULY 1977).

3. REDUCING SUGARS.

REMOVE CARBON DIOXIDE AS INDICATED IN METHOD NUMBER 1 AND PROCEED AS INDICATED IN THE OFFICIAL WINE METHODS NUMBERS 7 (A) AND 7 (B) (OFFICIAL STATE BULLETIN OF 23 JULY 1977) AND 7 (C) (OFFICIAL BULLETIN OF THE STATE ' OF 30 AUGUST 1979).

4. SULFUROUS ANHYDRIDE.

PROCEED AS INDICATED IN THE OFFICIAL METHOD OF WINES NUMBERS 28 (A) AND 28 (B) ("OFFICIAL STATE BULLETIN" OF JULY 26, 1977).

5. METHANOL.

REMOVE CARBON DIOXIDE AS INDICATED IN METHOD NUMBER 1 AND PROCEED AS INDICATED IN THE OFFICIAL METHOD OF WINE NUMBER 37 ("OFFICIAL STATE BULLETIN" OF 27 JULY 1977).

6. VOLATILE ACIDITY.

PROCEED AS INDICATED IN THE OFFICIAL METHOD OF WINE NUMBER 21 ("OFFICIAL STATE BULLETIN" OF 23 JULY 1977).

7. TOTAL DRY EXTRACT.

REMOVE CARBON DIOXIDE AS INDICATED IN METHOD NUMBER 1 AND PROCEED AS INDICATED IN THE OFFICIAL WINE METHOD NUMBER 6 ("OFFICIAL STATE BULLETIN" OF 23 JULY 1977).

8. ASHES.

REMOVE CARBON DIOXIDE AS INDICATED IN METHOD NUMBER 1 AND PROCEED AS INDICATED IN THE OFFICIAL METHOD OF WINE NUMBER 11 ("OFFICIAL STATE BULLETIN" OF 23 JULY 1977).

9. IRON.

REMOVE CARBON DIOXIDE AS INDICATED IN METHOD NUMBER 1 AND PROCEED AS INDICATED IN THE OFFICIAL METHOD OF WINE NUMBER 16 ("OFFICIAL STATE BULLETIN" OF 23 JULY 1977).

10. ETANAL.

1O.1. PRINCIPLE:

THE ETHANAL IS DETERMINED IN THE CIDER, PREVIOUSLY DISCOLOURED WITH COAL, MEASURING AT 570 NM, THE GREEN OR VIOLET COLORATION THAT GIVES WITH THE NITROPRUSATE AND PIPERIDINE.

10.2. MATERIAL AND APARATOS:

10.2.1. SPECTROPHOTOMETER CAPABLE OF PERFORMING READINGS AT 570 NM.

10.3. REAGENTS:

10.3.1. PIPERIDINE SOLUTION AT 10 PER 100 (V/V).

PREPARE BEFORE EACH MEASURE THE MIXTURE OF 2 ML. PIPERIDINE WITH 18 ML. DESTIL-WATER.

10.3.2. SODIUM NITROPRUSUATE SOLUTION AT 0.4 PER 100 (P/V).

IN A 250 ML GRADUATED FLASK, DISSOLVE 1 G. SODIUM NITROPRUSATE

(FORMULA OMITTED)

PULVERIZED. COMPLETE TO THE ROOT WITH DISTILLED WATER.

10.3.3. ACTIVE CARBON.

10.3.4. HYDROCHLORIC ACID DILUTED TO 25 PER 100 (V/V) (ABOUT 102 G/L.).

10.3.5. ALCALINE SOLUTION.

DISSOLVE 8.75 G OF BORIC ACID IN 400 ML. NORMAL SODIUM HYDROXIDE SOLUTION. COMPLETE UP TO ONE LITRE WITH DISTILLED WATER.

10.4. PROCEDURE:

10.4.1. DETERMINATION OF ETHANAL.

PLACE 25 ML. OF LIQUID TO BE ANALYZED IN A 100 ML ERLENMEYER, ADDING ABOUT 2 G. OF ACTIVATED CARBON. SHAKE VIGOROUSLY FOR A FEW SECONDS, LEAVE TO REST FOR TWO MINUTES, AND FILTER ON A FINE-PORE FOLD FILTER TO OBTAIN CLEAR FILTERING.

TWO ML. COLORLESS FILTERING IS PLACED IN A 100 ML ERLENMEYER, AND IS ADDED, AGITATING, 5 ML. SODIUM NITROPRUSUATE SOLUTION AND 5 ML. SOLUTION OF PIPERIDINE; MIX AND IMMEDIATELY PLACE THE MIXTURE IN THE 10 MM READING BUCKET. THICK. THE OBTAINED COLOURING, WHICH VARIES FROM GREEN TO VIOLET, IS MEASURED AT 570 NM. WITH REGARD TO AIR. THIS COLORING INCREASES, TO DECREASE QUICKLY. TO CHOOSE THE ULTIMATE VALUE OF THE MAXIMUM ABSORBENCY, WHICH IS OBTAINED AFTER ABOUT FIFTY SECONDS.

THE AMOUNT OF ETHANAL CONTENT IN THE ANALYZED LIQUID IS DEDUCED FROM A PATTERN CURVE.

10.4.2. PATTERN CURVE CONSTRUCTION.

ETHANAL PATTERN SOLUTION COMBINED WITH SULFUROUS ANHYDRIDE.

PREPARE A VALUED SOLUTION OF 5 TO 6 PER 100 (W/V) IN SO2 AND DETERMINE ITS EXACT AMOUNT BY MEASURING IT WITH AN IODINE-RATED SOLUTION O,1N.

IN A ONE-LITRE VOLUMETRIC FLASK, PLACE THE VOLUME OF THIS SOLUTION CORRESPONDING TO 1,500 MG. OF SO2. INSERT INTO THE FLASK, AIDED WITH A FUNNEL, AROUND 1 ML. RECENTLY DISTILLED ETHANAL AND COLLECTED ON A REFRIGERANT MIXTURE. MAKE UP TO ONE LITRE WITH DISTILLED WATER, MIX AND LEAVE THROUGHOUT THE NIGHT.

THE EXACT AMOUNT OF THIS ETHANAL PATTERN SOLUTION COMBINED WITH SO2 SHOULD BE DETERMINED AS FOLLOWS:

IN A 500 ML ERLENMEYER, PLACE 50 ML. ETHANAL PATTERN SOLUTION, ADDING 20 ML. OF THE DILUTED HYDROCHLORIC ACID AND 100 ML. DESTIL-WATER. OXIDIZE THE FREE SO2 WITH THE IODINE SOLUTION 0.1N IN THE PRESENCE OF STARCH IN THE FORM OF A WEAK BLUE COLOUR.

ADD A FEW DROPS OF PHENOLPHTHALEIN AND ENOUGH ALKALINE SOLUTION TO THE DISAPPEARANCE OF THE BLUE COLORING AND PINK APPEARANCE. RATE CO2 COMBINING ETHANAL WITH IODINE 0.1N UP TO WEAK BLUE COLORATION. WHERE N IS THE VOLUME SPENT, THE ETANAL STANDARD SOLUTION COMBINED WITH SO2 CONTAINS 44.05 N MG. OF ETHANAL PER LITRE.

PREPARATION OF THE PATTERN CURVE:

IN 100 ML GRADUATED FLASKS. PLACE SUCCESSIVELY 5-10-15 -20 AND 25 ML. OF THE STANDARD SOLUTION. MAKE UP WITH DISTILLED WATER. THESE DILUTIONS CORRESPOND TO QUANTITIES OF ETHANAL NEAR 40-80-120-160 AND 200 MG/L. THE EXACT QUANTITY OF THE DILUTIONS SHOULD BE CALCULATED FROM THE ETHANAL CONTENT OF THE PREVIOUSLY DETERMINED STANDARD SOLUTION.

PROCEED TO THE MEASURE OF ETHANAL OVER 2 ML. OF EACH OF THESE SOLUTIONS, AS INDICATED ABOVE. THE GRAPHIC REPRESENTATION OF THE ABSORBENCES OF THESE SOLUTIONS ACCORDING TO THE QUANTITY OF ETHANAL IS A STRAIGHT LINE THAT DOES NOT PASS THROUGH THE ORGIEN.

10.5. CALCULATIONS:

CALCULATE THE CONTENT IN ETHANAL BY COMPARISON WITH THE CORRESPONDING PATTERN CURVE.

10.6. REMARKS:

IF, EXCEPTIONALLY, THE LIQUID TO BE ANALYZED CONTAINS FREE ETHANAL, NOT COMBINED WITH THE SO2, IT WILL BE NECESSARY BEFORE STARTING TO DETERMINE THE TOTAL ETHANAL PASS BEFORE THE STATE OF COMBINATION WITH THE SO2. TO DO THIS, ADD A PART OF THE LIQUID TO ANALYZE A SMALL EXCESS OF FREE SO2 AND WAIT A FEW HOURS BEFORE PROCEEDING TO THE DETERMINATION.

10.7. REFERENCES:

1. H. REBELLION (1970) DTSCH. LEBENSMIT. RDSCH 1970, 66, 5-6.

2. RECUEIL DES METHODS INTERNATIONALES D' ANALYSE DES VINS. O.I.V. A-37, 1978.

11. CARBONIC ANHYDRIDE.

11.1. PRINCIPLE:

AFTER THE COLD OXIDATION OF THE SULPHUR DIOXIDE BY THE OXYGENATED WATER IN THE PRESENCE OF THE CUPRIC ION, THE CARBON DIOXIDE DISPLACED FROM THE CIDER BY A BUFFER SOLUTION OF PHOSPHATE AND PHOSPHORIC ACID IS COLD-DRAGGED IN CIRCUIT CLOSED BY A CURRENT OF AIR BY MEANS OF A SMALL CIRCULATION PUMP TOWARDS A STANDARD SOLUTION OF BARIUM HYDROXIDE. THE EXCESS OF THIS BASE IS IMMEDIATELY DETERMINED WITH A STANDARD SOLUTION OF HYDROCHLORIC ACID IN THE PRESENCE OF A MIXED INDICATOR OF TIMOLPHTHALEINA-PHENOLPHTHALEIN.

APPLICABLE TO CONTENTS LESS THAN 6 G/L.

11.2. MATERIAL AND APARATOS:

11.2.1. GLASS APPARATUS FOR EXTRANCUTION OF CO2 IN CLOSED CIRCUIT (FIGURE 1), COMPOSITE OF:

-A 250 ML MATRAZ. (1) FITTED WITH AN ANTI-FOAM BALL ON WHICH A BURETTE (5) IS ADAPTED;

-TWO BARS (2) AND (3);

-A PUMP (4) THAT ENSURES AIR CIRCULATION IN THE CIRCUIT (2 TO 10 BUBBLES PER SECOND).

(FIGURE 1 OMITIDA)

11.3. REAGENTS:

11.3.1. SODIUM HYDROXIDE SOLUTION O,2N.

11.3.2. BARIUM CHLORIDE SOLUTION, WITH 90 GR. OF CL2BA2. 2H20 PER LITRO.

11.3.3. BARIC HYDROXIDE SOLUTION.

MIX TWO VOLUMES OF THE 90 G/L BARIC CHLORIDE SOLUTION WITH A VOLUME OF SODIUM HYDROXIDE O,2N SOLUTION.

STORE IN A CONTAINER THAT PREVENTS CARBONATION.

11.3.4. TAMPON SOLUTION.

DISSOLVE IN RECENTLY DISTILLED WATER BOILED AT 400 G. MONOSODIUM PHOSPHATE (FORMULA OMITTED) AND ADD 50 ML. PHOSPHORIC ACID (FORMULA OMITTED) AND COMPLETE UP TO 1 L. WITH WATER ML. PHOSPHORIC ACID (OMITTED FORMULA) AND RECENTLY BOILED DISTILLED.

11.3.5. SODIUM HYDROXIDE AT 50 PER 100 (P/P).

AQUEOUS SOLUTION (O= 1,525) THAT OCNHAS 50 G. SODIUM HYDROXIDE IN 100 G SOLUTION.

USE FIFTEEN DAYS AFTER PREPARATION TO ACHIEVE THE EVENTUAL DECANTATION (FORMULA OMITTED). THIS SOLUTION MUST BE TRANSPARENT.

11.3.6 OXYGENATED WATER IN 10-VOLUME SOLUTION. THIS SOLUTION CONTAINS 30.4 G OF (FORMULA OMITTED) PER LITER AND RELEASES 10 TIMES ITS VOLUME OF OXYGEN BY CATALYTIC DECOMPOSITION WITH (FORMULA OMITTED) IN ALKALINE MEDIUM.

11.3.7. COPPER SLUT (FORMULA OMITTED) Pulverized.

11.3.8. DISSOLVE 1 G. OF PHENOLFTLALEINE AND O, 5 G. OF TIMOLPHTHALEIN NEUTRAL ALCOHOL OF 96 PER 100 AND COMPLETE UP TO 100 ML.

11.3.9. HYDROCHLORIC ACID O,2N.

11.4 PROCEDURE:

11.4.1. SIDRAS CONTAINING 4 G OF CO 2 G PER LITRE.

THE CIDER BOTTLE SHOULD BE KEPT IN A HORIZONTAL POSITION FOR A NIGHT TO OR DEGREES C.

PLACE IN THE BARBOTER (2) 150 ML. EXACTLY MEASURED BARIUM HYDROXIDE SOLUTION.

INTRODUCE INTO THE FLASK (1) SOME CRYSTALS OF COPPER SULPHATE AND 5 ML. 10-VOLUME OXYGENATED WATER. CAREFULLY PUT IN A FLASK, PREVIOUSLY RINSED AND COOLED TO OR DEGREES C, 50 ML. FROM SIDRA. PLACE THE FLASK (1) IN THE CIRCUIT AND WAIT TEN MINUTES.

CONNECT THE PUMP AND ENTER THROUGH THE BURETA (5) 20ML. TAMPON SOLUTION. LEAVE IN OPERATION FOR THIRTY MINUTES.

STOP THE PUMP AND TAKE FROM THE BARBOTTER (2) 50 ML. OF THE BARIC HYDROXIDE SOLUTION; VALUE IT WITH HYDROCHLORIC ACID O,2N WITH MIXED INDICATOR. BE V TWO THE ML NUMBER. SPENT.

IT IS ESSENTIAL TO ASSESS THE BARIC HYDROXIDE DAILY USING 150 ML. BE V ONE THE NUMBER OF MILLILITRES OF HYDROCHLORIC ACID O,2N NEEDED.

11.4.2. SIDRAS WHICH CONTINUE TO BE MORE THAN 4 G OF CO 2 PER LITRE.

THE BOTTLE SHOULD BE KEPT IN A HORIZONTAL POSITION TO OR DEGREES C FOR ONE NIGHT. OPEN THE BOTTLE WITH CAUTION. FOR 75O ML BOTTLES. SUBSTITUTED 5O ML. OF SIDE BY 50 ML. SODIUM HYDROXIDE AT 50 PER 100, GENTLY SLIPPING THE SOLUTION BY THE NECK OF THE BOTTLE. CLOSE IT IS IMMEDIATELY AND HOMOGENIZE ITS CONTENTS BY STIRRING IT UP; THEN PLACE IT IN A WATER BATH AT 20 DEGREES C.

PLACE ON THE BARBOTER (2) 300 ML. ACCURATE SOLUTION OF BARIC HYDROXIDE.

INTRODUCE INTO THE FLASK (1) COPPER SULPHATE CRYSTALS AND 5 ML. 10-VOLUME OXYGENATED WATER.

SLOWLY ENTER THE PIPETTE INTO THE BOTTLE TO AVOID A SQUARE BLOCK AND TAKE 50 ML. FROM THE CIDER AND CAST THEM INTO THE FLASK. ATTACH IT TO THE APPLIANCE AND LET IT REST TEN MINUTES.

THE VOLUME OF THE CIDER TAKEN SHOULD BE REDUCED TO 25 ML. IF IT IS MORE THAN 8 G/L. CARBONIC ANHYDRIDE.

CONNECT THE PUMP AND ENTER THE FLASK (1) WITH A BURETTE (5), 40 ML. TAMPON SOLUTION. PUT INTO OPERATION FOR THIRTY MINUTES.

STOP THE PUMP AND TAKE FROM THE BARBOTTER (2) 50 ML. OF THE BARIC HYDROXIDE SOLUTION. VALUE IT WITH HYDROCHLORIC ACID O,2N IN THE PRESENCE OF MIXED INDICATOR. BE V SQUARE THE NUMBER OF ML. USED. BE V ONE THE ML NUMBER. HYDROCHLORIC ACID O,2N CONSUMED TO VALUE 150 ML. BARIC HYDROXIDE SOLUTION.

11.5. CALCULATIONS:

11.5.1. FOR SSIDAS THAT CONTINUE TO BE LESS THAN 4 G OF CO 2 PER LITRE:

CO 2 AMOUNTS IN GRAMS PER LITRE IN CIDER

(FORMULA OMITTED)

11.5.2. FOR SIDRAS WHICH ARE NOT MORE THAN 4 G OF CO 2 PER LITRE:

MEASURE THE VOLUME OF LIQUID LEFT IN THE BOTTLE AND ADD 50 OR 25 ML. FROM THE SAMPLE TAKEN FOR DETERMINATION. BE V THE TOTAL VOLUME SO DETERMINED. THEN CALCULATE THE CORRECTION FACTOR F, WHICH IS NECESSARY TO TAKE INTO ACCOUNT THE 50 ML. SODIUM HYDROXIDE 50 PER 100 INTRODUCED INTO THE BOTTLE.

(FORMULA OMITTED)

IF THE SAMPLE FOR DETERMINATION IS 50 ML., THE AMOUNT OF CARBON DIOXIDE IN GR/L. ES (FORMULA OMITTED).

THIS RESULT MUST BE MULTIPLIED BY 2 IF THE SAMPLE INTAKE IS 25 ML.

11.6. BIBLIOGRAPHY:

1. JAULMES (P): METH. AN. 1972, NUM. 422; ANN. FALS. EXP. CHIM., 1973 NUM. 708, 96-109. 2. RECUEIL DES METHODS INTERNATIONALES D' ANALYSE DES VINS O.I.V.I. A-39, 1978.