Commission Regulation (EC) No 1082/96 of 14 June 1996 establishing a reference method for the determination of the ethyl ester of beta-apo-8' carotenic acid in concentrated butter and butter
Modified by
  • Commission Regulation (EC) No 175/1999of 26 January 1999amending Regulations (EEC) No 3942/92, (EC) No 86/94, (EC) No 1082/96 and (EC) No 1459/98 establishing reference methods for the determination of certain tracers in butter, butteroil and cream, 31999R0175, January 27, 1999
  • Commission Regulation (EC) No 213/2001of 9 January 2001laying down detailed rules for the application of Council Regulation (EC) No 1255/1999 as regards methods for the analysis and quality evaluation of milk and milk products and amending Regulations (EC) No 2771/1999 and (EC) No 2799/1999, 32001R0213, February 7, 2001
Commission Regulation (EC) No 1082/96of 14 June 1996establishing a reference method for the determination of the ethyl ester of beta-apo-8′ carotenic acid in concentrated butter and butter THE COMMISSION OF THE EUROPEAN COMMUNITIES,Having regard to the Treaty establishing the European Community,Having regard to Council Regulation (EEC) No 804/68 of 27 June 1968 on the common organization of the market in milk and milk productsOJ No L 148, 28. 6. 1968, p. 13., as last amended by Commission Regulation (EC) No 2931/95OJ No L 307, 20. 12. 1995, p. 10., and in particular Article 6 (7) and Article 12 (3) thereof,Whereas Commission Regulation (EEC) No 570/88 of 16 February 1988 on the sale of butter at reduced prices and the granting of aid for butter and for concentrated butter for use in the manufacture of pastry products, ice-cream and other foodstuffsOJ No L 55, 1. 3. 1988, p. 31., as last amended by Regulation (EC) No 531/96OJ No L 78, 28. 3. 1996, p. 13., provides for the tracing of the subsidized butter and concentrated butter in certain circumstances in order to ensure the correct end use of these products;Whereas, in view of the importance of tracing to the proper functioning of the scheme and in order to ensure the equal treatment of operators who participate in it, it is appropriate to establish common methods for the determination of the tracers referred to in Regulation (EEC) No 570/88;Whereas it is difficult to establish such reference methods for all tracers simultaneously; whereas establishing a reference method for the determination of the ethyl ester of beta-apo-8′ carotenic acid in concentrated butter or butter constitutes a further step in this direction;Whereas the measures provided for in this Regulation are in accordance with the opinion of the Management Committee for Milk and Milk Products,HAS ADOPTED THIS REGULATION:
Article 1The reference method of analysis specified in the Annex shall be applied for the determination of the beta-apo-8′ carotenic acid ethyl ester content of concentrated butter or butter under Regulation (EC) No 2571/97.Concentrated butter or butter have been traced in conformity with Article 6 of Regulation (EC) No 2571/97 if the results obtained are in accordance with the specifications of paragraph 8 of the Annex.
Article 2This Regulation shall enter into force on the third day following its publication in the Official Journal of the European Communities.It shall apply from 1 October 1996.
This Regulation shall be binding in its entirety and directly applicable in all Member States.ANNEXDETERMINATION OF THE ETHYL ESTER OF BETA-APO-8′-CAROTENIC ACID IN CONCENTRATED BUTTER AND BUTTER BY SPECTROMETRY1.Scope and field of applicationThe method describes a procedure for the quantitative determination of the ethyl ester of beta-apo-8′-carotenic acid (apo carotenic ester) in concentrated butter and butter. The apo carotenic ester is the sum of all substances present in an extract of samples obtained under the conditions described in the method which absorb light at 440 nm. It is applicable to samples received under Regulation (EEC) No 570/88.2.PrincipleThe butterfat is dissolved in light petroleum and the absorbance measured at 440 nm. The apo carotenic ester content is determined by reference to an external standard.3.Apparatus3.1.Pipettes — graduated, of capacity 0,25, 0,50, 0,75 and 1,0 ml3.2.Spectrophotometer — suitable for use at 440 nm (and 447 — 449 nm) and equipped with cells of optical path length 1 cm3.3.Volumetric flasks e.g. 20 ml and 100 ml4.ReagentsAll reagents must be of recognized analytical grade.4.1.Apo carotenic ester suspension (approximately 20 %)4.1.1.Establish the content of the suspension as follows:weigh accurately approximately 400 mg into a 100 ml volumetric flask and dissolve in petroleum spirit (4.2), dilute to volume with petroleum spirit. Dilute 5,0 ml of this solution to 100 ml with petroleum spirit. (Solution A.) Dilute 5,0 ml of Solution A to 100 ml with petroleum spirit. Measure the absorbance at 447 — 449 nm (measure the maximum against petroleum spirit as a blank using 1 cm cells).Apo carotenic ester content (%)Amax · 40 000A · 2 550A maxabsorbance of the measuring solution at the maximumAweight of sample (g)2550reference A (1 %, 1 cm) valueThe purity of the suspension is P (%).Note: Apo carotenic ester suspension is sensitive to air, heat and light. In the unopened, original container (sealed under nitrogen) and in a cool place it can be stored for about 12 months. After opening the contents should be used within a short period.4.1.2.Apo carotenic ester standard solution, approx 0,2 mg/mlWeigh to the nearest 0,1 mg about 0,100 g of apo carotenic ester suspension (4.1.1) (Wg), dissolve in petroleum spirit (4.2), transfer quantitatively into a volumetric flask of capacity 100 ml, and make up to the mark with petroleum spirit.This solution contains (W.P)/100 mg/ml of apo carotenic ester.Note: The solution must be stored in a cool place in the dark. Discard unused solution after one month.4.2.Petroleum spirit (40 — 60 °C).4.3.Sodium sulphate, anhydrous, granular previously dried at 102 °C for two hours.5.Procedure5.1.Preparation of the test sample5.1.1.Concentrated butterMelt the sample in an oven at approximately 45 °C.5.1.2.ButterMelt the sample in an oven at approximately 45 °C and filter a representative portion through a filter containing about 10 g of anhydrous sodium sulphate (4.3) in an environment shielded from strong natural and artificial light and maintained at 45 °C. Collect a suitable amount of butterfat.5.2.DeterminationWeigh, to the nearest 1 mg approximately 1 g of concentrated butter, (or extracted butterfat (5.1.2)), (mg). Transfer quantitatively to a 20 ml (Vml) volumetric flask using petroleum spirit (4.2), make up to the mark and mix thoroughly.Transfer an aliquot to a 1 cm cell and measure the absorbance at 440 nm, against a petroleum spirit blank. Obtain the concentration of apo carotenic ester in the solution by reference to the calibration graph (C μg/ml).5.3.Calibration graphPipette 0, 0,25, 0,5, 0,75 and 1,0 ml of apo carotenic ester standard solution (4.1.2) into five 100 ml volumetric flasks. Dilute to volume with petroleum spirit (4.2) and mix.The approximate concentrations of the solutions range from 0 to 2 μg/ml and are calculated accurately by reference to the concentration of the standard solution (4.1.2) (W.P)/100 mg/ml. Measure the absorbances at 440 nm against a petroleum spirit (4.2) blank.Plot the values of absorbance on the y axis against apo carotenic ester concentration on the x axis.6.Calculation of results6.1.Apo carotenic ester content, expressed as mg/kg product is given by:Concentrated butter: (C.V)/mButter: 0,82 (C.V)/mwhereCapo carotenic ester content, μg/ml, read from the calibration graph (5.3),mmass (g) of the test portion (5.2),0,82a correction factor for the butterfat content of butter.7.Accuracy of the method7.1.Repeatability7.1.1.Butter analysisThe difference between the results of two determinations carried out within the shortest feasible time interval, by one operator using the same apparatus on identical test material shall not exceed 1,4 mg/kg.7.1.2.Concentrated butter analysisThe difference between the results of two determinations carried out within the shortest feasible time interval, by one operator using the same apparatus on identical test material shall not exceed 1,6 mg/kg.7.2.Reproducibility7.2.1.Butter analysisThe difference between the results of two determinations carried out by operators in different laboratories, using different apparatus on identical test material shall not exceed 4,7 mg/kg.7.2.2.Concentrated butter analysisThe difference between the results of two determinations carried out by operators in different laboratories, using different apparatus on identical test material shall not exceed 5,3 mg/kg.7.3.Source of precision dataThe precision data were determined from an experiment conducted in 1995 involving 11 laboratories and 12 traced samples (six blind duplicates) for butter; 12 traced (six blind duplicates) for concentrated butter.8.Tolerance limits8.1.Three samples must be taken from the traced product in order to check on the correct tracing of the product.8.2.Butter8.2.1.The incorporation rate for butter, taking into account background absorbance, is 22 mg/kg.8.2.2. The results of the three samples obtained from the analysis of the product is used to check the rate and the homogeneity of tracer incorporation and the lowest of these results is compared with the following limits (Critical Difference for a 95 % probability level (CrD95) taken into consideration):18,0 mg/kg (95 % of the minimum incorporation rate),13,0 mg/kg (70 % of the minimum incorporation rate).The tracer concentration of the sample giving the lowest result is used in conjunction with interpolation between 18,0 mg/kg and 13,0 mg/kg.8.3.Concentrated butter8.3.1.The incorporation rate for concentrated butter, taking into account background absorbance, is 24 mg/kg.8.3.2. The results of the three samples obtained from the analysis of the product is used to check the rate and the homogeneity of tracer incorporation and the lowest of these results is compared with the following limits (Critical Difference for a 95 % probability level (CrD95) taken into consideration):20,0 mg/kg (95 % of the minimum incorporation rate),14,0 mg/kg (70 % of the minimum incorporation rate).The tracer concentration of the sample giving the lowest result is used in conjunction with interpolation between 20,0 mg/kg and 14,0 mg/kg.