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Commission Regulation (EC) No 273/2008 (repealed)Show full title

Commission Regulation (EC) No 273/2008 of 5 March 2008 laying 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Regulation (EC) No 1255/1999 as regards methods for the analysis and quality evaluation of milk and milk products (repealed)

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  1. Introductory Text

  2. CHAPTER I GENERAL PROVISIONS

    1. Article 1.Subject matter and scope

    2. Article 2.Routine methods

  3. CHAPTER II METHODS OF ANALYSIS

    1. Article 3.Evaluation of compliance of a consignment with a legal limit

    2. Article 4.Sensory evaluation

    3. Article 5.Tracers

    4. Article 6.Detection of cows’ milk casein

    5. Article 7.Detection of coliforms

    6. Article 8.Determination of the lactose content

    7. Article 9.Detection of rennet whey

    8. Article 10.Detection of buttermilk

    9. Article 11.Detection of antimicrobiotic residues

    10. Article 12.Determination of skimmed-milk powder content

    11. Article 13.Detection of starch

    12. Article 14.Determination of moisture content in dried cream

    13. Article 15.Determination of moisture content of acid buttermilk powder

    14. Article 16.Determination of milk fat purity

  4. CHAPTER III GENERAL AND FINAL PROVISIONS

    1. Article 17.Quality assurance

    2. Article 18.Sampling and disputes over the results of analysis

    3. Article 19.Transition period

    4. Article 19a.Notifications

    5. Article 20.Repeals

    6. Article 21.Entry into force

  5. Signature

    1. ANNEX I

      (Article 1)

      1. LIST OF REFERENCE METHODS

    2. ANNEX II

      (Article 3)

      1. EVALUATION OF COMPLIANCE OF A CONSIGNMENT WITH THE LEGAL LIMIT...

        1. 1. PRINCIPLE

        2. 2. THE METHOD IS VALIDATED THROUGH INTERNATIONAL COLLABORATION

        3. 3. IN-HOUSE VALIDATION WITH CALCULATION OF INTERNAL REPRODUCIBILITY STANDARD DEVIATION

    3. ANNEX III

      (Article 4)

      1. EVALUATION OF ASSESSORS AND THE RELIABILITY OF RESULTS IN SENSORY...

        2. A. DETERMINATION OF THE ‘REPEATABILITY INDEX’

        3. B. DETERMINING THE ‘DEVIATION INDEX’

        4. C. COMPARISON OF THE RESULTS OBTAINED IN DIFFERENT REGIONS OF A...

    4. ANNEX IV

      (Article 4)

      1. SENSORY EVALUATION OF BUTTER

        1. 1. SCOPE

        2. 2. DEFINITIONS

        3. 3. TEST ROOM

        4. 4. SELECTION OF ASSESSORS

          2. 4.1. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

        5. 5. REQUIREMENTS FOR THE PANEL

        6. 6. PERFORMANCE

        7. 7. SAMPLING AND PREPARATION OF THE SAMPLE

        8. 8. ASSESSMENT OF THE VALUE OF EACH ATTRIBUTE

          1. 8.1. . . . . . . . . . ....

          2. 8.2. . . . . . . . . . ....

          3. 8.3. . . . . . . . . . ....

          4. 8.4. . . . . . . . . . ....

        9. 9. SUPERVISION

        10. 10. NOMENCLATURE

        11. 11. REFERENCE

    5. ANNEX V

      (Article 5)

      1. DETERMINATION OF THE CONTENT OF ENANTHIC ACID TRIGLYCERIDE IN BUTTER,...

        1. 1. SCOPE

        2. 2. TERMS AND DEFINITION

        3. 3. PRINCIPLE

        4. 4. REAGENTS

          2. 4.1. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

          4. 4.3. . . . . . . . . . ....

          5. 4.4. . . . . . . . . . ....

        5. 5. APPARATUS

          2. 5.1. . . . . . . . . . ....

          3. 5.2. . . . . . . . . . ....

          4. 5.3. . . . . . . . . . ....

          5. 5.4. . . . . . . . . . ....

          6. 5.5. . . . . . . . . . ....

          7. 5.6. . . . . . . . . . ....

          8. 5.7. . . . . . . . . . ....

            2. 5.7.1. . . . . . . . . . ....

            3. 5.7.2. . . . . . . . . . ....

              2. 5.7.2.1. . . . . . . . . . ....

              3. 5.7.2.2. . . . . . . . . . ....

          9. 5.8. . . . . . . . . . ....

        6. 6. SAMPLING

        7. 7. PROCEDURE

          1. 7.1. Preparation of the test sample and test portion

            2. 7.1.1. Butter-oil, Butter

              1. 7.1.1.1. . . . . . . . . . ....

              2. 7.1.1.2. . . . . . . . . . ....

              3. 7.1.1.3. . . . . . . . . . ....

            3. 7.1.2. Cream

              1. 7.1.2.1. . . . . . . . . . ....

              2. 7.1.2.2. . . . . . . . . . ....

              3. 7.1.2.3. . . . . . . . . . ....

              4. 7.1.2.4. . . . . . . . . . ....

              5. 7.1.2.5. . . . . . . . . . ....

              6. 7.1.2.6. . . . . . . . . . ....

          2. 7.2. Preparation of the calibration standards

            1. 7.2.1. . . . . . . . . . ....

            2. 7.2.2. . . . . . . . . . ....

            3. 7.2.3. . . . . . . . . . ....

            4. 7.2.4. . . . . . . . . . ....

            5. 7.2.5. . . . . . . . . . ....

          3. 7.3. Chromatographic determination

            1. 7.3.1. . . . . . . . . . ....

            2. 7.3.2. . . . . . . . . . ....

            3. 7.3.3. . . . . . . . . . ....

        8. 8. CALCULATION OF RESULTS

          2. 8.1. Calibration

            1. 8.1.1. . . . . . . . . . ....

            2. 8.1.2. . . . . . . . . . ....

            3. 8.1.3. . . . . . . . . . ....

          3. 8.2. Test samples

        9. 9. PRECISION

          2. 9.1. Repeatability

          3. 9.2. Reproducibility

        10. 10. TOLERANCE LIMITS: LOWER LIMITS (CASE OF INSUFFICIENT QUANTITIES)

          1. 10.1. . . . . . . . . . ....

          2. 10.2. Butter and concentrated butter

            1. 10.2.1. . . . . . . . . . ....

            2. 10.2.2. . . . . . . . . . ....

          3. 10.3. Cream

            1. 10.3.1. . . . . . . . . . ....

            2. 10.3.2. . . . . . . . . . ....

        11. 11. TOLERANCE LIMITS: UPPER LIMITS (CASE OF EXCEEDING QUANTITY BY MORE...

          1. 11.1. . . . . . . . . . ....

          2. 11.2. Butter and concentrated butter

            1. 11.2.1. . . . . . . . . . ....

          3. 11.3. Cream

            1. 11.3.1. . . . . . . . . . ....

        12. 12. ADDITIONAL INFORMATION: STATISTICAL ANALYSIS OF RESULTS ON THE DETERMINATION OF...

          2. Tests for outliers:

          3. Precision parameters

          4. Figure 1

            1. Experimental results: Sample A

            2. Experimental results: Sample B

            3. Experimental results: Sample C

            4. Experimental results: Sample D

    6. ANNEX VI

      (Article 5)

      1. DETERMINING THE VANILLIN CONTENT IN CONCENTRATED BUTTER, BUTTER OR CREAM...

        1. 1. SCOPE AND FIELD OF APPLICATION

        2. 2. PRINCIPLE

        3. 3. APPARATUS

          2. 3.1. . . . . . . . . . ....

          3. 3.2. . . . . . . . . . ....

          4. 3.3. . . . . . . . . . ....

          5. 3.4. . . . . . . . . . ....

          6. 3.5. . . . . . . . . . ....

          7. 3.6. . . . . . . . . . ....

          8. 3.7. . . . . . . . . . ....

        4. 4. REAGENTS

          2. 4.1. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

          4. 4.3. . . . . . . . . . ....

          5. 4.4. . . . . . . . . . ....

          6. 4.5. . . . . . . . . . ....

            2. 4.5.1. Vanillin stock solution (= 500 μg/ml)

            3. 4.5.2. Vanillin standard solution (= 10 μg/ml)

            4. 4.5.3. . . . . . . . . . ....

            5. 4.5.4. . . . . . . . . . ....

            6. 4.5.5. . . . . . . . . . ....

            7. 4.5.6. HPLC mobile phase

        5. 5. PROCEDURE

          1. 5.1. Preparation of the test sample

            1. 5.1.1. Butter

            2. 5.1.2. Concentrated butter

            3. 5.1.3. Cream

          2. 5.2. Preparation of the test solution

          3. 5.3. Calibration

          4. 5.4. Determination by HPLC

        6. 6. CALCULATION OF THE RESULTS

        7. 7. ACCURACY OF THE METHOD

          1. 7.1. Repeatability (r)

          2. 7.2. Reproducibility (R)

        8. 8. TOLERANCE LIMITS

          1. 8.1. . . . . . . . . . ....

          2. 8.2. . . . . . . . . . ....

            2. 8.2.1. . . . . . . . . . ....

            3. 8.2.2. . . . . . . . . . ....

          3. 8.3. . . . . . . . . . ....

            2. 8.3.1. . . . . . . . . . ....

            3. 8.3.2. . . . . . . . . . ....

        9. 9. NOTES

          1. 9.1. . . . . . . . . . ....

          2. 9.2. . . . . . . . . . ....

          3. 9.3. . . . . . . . . . ....

          4. 9.4. . . . . . . . . . ....

    7. ANNEX VII

      (Article 5)

      1. DETERMINING THE ETHYL ESTER OF BETA-APO-8'-CAROTENIC ACID IN CONCENTRATED BUTTER...

        1. 1. SCOPE AND FIELD OF APPLICATION

        2. 2. PRINCIPLE

        3. 3. APPARATUS

          1. 3.1. . . . . . . . . . ....

          2. 3.2. . . . . . . . . . ....

          3. 3.3. . . . . . . . . . ....

          4. 3.4. . . . . . . . . . ....

          5. 3.5. . . . . . . . . . ....

          6. 3.6. . . . . . . . . . ....

        4. 4. REAGENTS

          2. 4.1. . . . . . . . . . ....

            2. 4.1.1. . . . . . . . . . ....

            3. 4.1.2. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

          4. 4.3. . . . . . . . . . ....

          5. 4.4. . . . . . . . . . ....

          6. 4.5. . . . . . . . . . ....

        5. 5. PROCEDURE

          1. 5.1. Preparation of the test sample

            1. 5.1.1. Concentrated butter

            2. 5.1.2. Butter

          2. 5.2. Determination

          3. 5.3. Calibration

        6. 6. CALCULATION OF THE RESULTS

          1. 6.1. . . . . . . . . . ....

        7. 7. ACCURACY OF THE METHOD

          1. 7.1. Repeatability

            1. 7.1.1. Butter analysis

            2. 7.1.2. Concentrated butter analysis

          2. 7.2. Reproducibility

            1. 7.2.1. Butter analysis

          3. 7.3. Concentrated butter analysis

          4. 7.4. Source of precision data

        8. 8. TOLERANCE LIMITS

          1. 8.1. . . . . . . . . . ....

          2. 8.2. Butter

            1. 8.2.1. . . . . . . . . . ....

            2. 8.2.2. . . . . . . . . . ....

          3. 8.3. Concentrated butter

            1. 8.3.1. . . . . . . . . . ....

    8. ANNEX VIII

      (Article 5)

      1. DETERMINING SITOSTEROL OR STIGMASTEROL IN BUTTER OR CONCENTRATED BUTTER BY...

        1. 1. SCOPE AND FIELD OF APPLICATION

        2. 2. PRINCIPLE

        3. 3. APPARATUS

          1. 3.1. . . . . . . . . . ....

          2. 3.2. . . . . . . . . . ....

          3. 3.3. . . . . . . . . . ....

          4. 3.4. . . . . . . . . . ....

          5. 3.5. . . . . . . . . . ....

          6. 3.6. . . . . . . . . . ....

          7. 3.7. . . . . . . . . . ....

          8. 3.8. . . . . . . . . . ....

            2. 3.8.1. . . . . . . . . . ....

            3. 3.8.2. . . . . . . . . . ....

            4. 3.8.3. . . . . . . . . . ....

            5. 3.8.4. . . . . . . . . . ....

          9. 3.9. . . . . . . . . . ....

          10. 3.10. . . . . . . . . . ....

        4. 4. REAGENTS

          2. 4.1. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

          4. 4.3. . . . . . . . . . ....

            2. 4.3.1. . . . . . . . . . ....

              2. 4.3.1.1. . . . . . . . . . ....

              3. 4.3.1.2. . . . . . . . . . ....

          5. 4.4. . . . . . . . . . ....

          6. 4.5. . . . . . . . . . ....

          7. 4.6. . . . . . . . . . ....

          8. 4.7. . . . . . . . . . ....

          9. 4.8. . . . . . . . . . ....

            2. 4.8.1. . . . . . . . . . ....

          10. 4.9. . . . . . . . . . ....

            2. 4.9.1. . . . . . . . . . ....

          11. 4.10. . . . . . . . . . ....

        5. 5. METHOD

          1. 5.1. Preparation of standard solutions for chromatography

            2. 5.1.1. . . . . . . . . . ....

            3. 5.1.2. . . . . . . . . . ....

          2. 5.2. Preparation of the unsaponifiables

            1. 5.2.1. . . . . . . . . . ....

            2. 5.2.2. . . . . . . . . . ....

            3. 5.2.3. . . . . . . . . . ....

          3. 5.3. Preparation of trimethyl silyl ethers

            1. 5.3.1. . . . . . . . . . ....

            2. 5.3.2. . . . . . . . . . ....

          4. 5.4. Gas-chromatographic analysis

            1. 5.4.1. Choice of operating conditions

            2. 5.4.2. Analytical procedure

        6. 6. CALCULATION OF RESULTS

          1. 6.1. . . . . . . . . . ....

        7. 7. ACCURACY OF THE METHOD

          1. 7.1. Butter

            1. 7.1.1. Repeatability

              1. 7.1.1.1. Stigmasterol

              2. 7.1.1.2. Sitosterol

            2. 7.1.2. Reproducibility

              1. 7.1.2.1. Stigmasterol

              2. 7.1.2.2. Sitosterol

            3. 7.1.3. Source of precision data

          2. 7.2. Concentrated butter

            1. 7.2.1. Repeatability

              1. 7.2.1.1. Stigmasterol

              2. 7.2.1.2. Sitosterol

            2. 7.2.2. Reproducibility

              1. 7.2.2.1. Stigmasterol

              2. 7.2.2.2. Sitosterol

            3. 7.2.3. Source of precision data

        8. 8. TOLERANCE LIMITS

          1. 8.1. . . . . . . . . . ....

          2. 8.2. Butter

            1. 8.2.1. Stigmasterol

              1. 8.2.1.1. . . . . . . . . . ....

              2. 8.2.1.2. . . . . . . . . . ....

            2. 8.2.2. Sitosterol

              1. 8.2.2.1. . . . . . . . . . ....

              2. 8.2.2.2. . . . . . . . . . ....

          3. 8.3. Concentrated butter

            1. 8.3.1. Stigmasterol

              1. 8.3.1.1. . . . . . . . . . ....

              2. 8.3.1.2. . . . . . . . . . ....

            2. 8.3.2. Sitosterol

              1. 8.3.2.1. . . . . . . . . . ....

              2. 8.3.2.2. . . . . . . . . . ....

    9. ANNEX IX

      (Article 6)

      1. REFERENCE METHOD FOR THE DETECTION OF COWS’ MILK AND CASEINATE...

        1. 1. SCOPE

        2. 2. FIELD OF APPLICATION

        3. 3. PRINCIPLE OF THE METHOD

          1. 3.1. . . . . . . . . . ....

          2. 3.2. . . . . . . . . . ....

          3. 3.3. . . . . . . . . . ....

          4. 3.4. . . . . . . . . . ....

        4. 4. REAGENTS

          2. Isoelectric focusing

            1. 4.1. Reagents for production of the urea containing polyacrylamide gels

              1. 4.1.1. Stock gel solution

              2. 4.1.2. Gel solution

              3. 4.1.3. Catalyst solutions

                1. 4.1.3.1. . . . . . . . . . ....

                2. 4.1.3.2. . . . . . . . . . ....

            2. 4.2. Contact fluid

            3. 4.3. Anode solution

            4. 4.4. Cathode solution

          3. Sample preparation

            1. 4.5. Reagents for protein isolation

              1. 4.5.1. . . . . . . . . . ....

              2. 4.5.2. . . . . . . . . . ....

              3. 4.5.3. . . . . . . . . . ....

            2. 4.6. Protein dissolving buffer

            3. 4.7. Reagents for plasmin cleavage of caseins

              1. 4.7.1. Ammonium carbonate buffer

              2. 4.7.2. . . . . . . . . . ....

              3. 4.7.3. ε-Aminocaproic acid solution for enzyme inhibition

            4. 4.8. Standards

              1. 4.8.1. . . . . . . . . . ....

              2. 4.8.2. Preparation of laboratory interim-standards of buffalos’ renneted milk containing 0 %...

          4. Reagents for protein staining

            1. 4.9. Fixative

            2. 4.10. Destaining solution

            3. 4.11. Staining solutions

              1. 4.11.1. Staining solution (stock solution 1)

              2. 4.11.2. Staining solution (stock solution 2)

              3. 4.11.3. Staining solution (working solution)

        5. 5. EQUIPMENT

          1. 5.1. . . . . . . . . . ....

          2. 5.2. . . . . . . . . . ....

          3. 5.3. . . . . . . . . . ....

          4. 5.4. . . . . . . . . . ....

          5. 5.5. . . . . . . . . . ....

          6. 5.6. . . . . . . . . . ....

          7. 5.7. . . . . . . . . . ....

          8. 5.8. . . . . . . . . . ....

          9. 5.9. . . . . . . . . . ....

          10. 5.10. . . . . . . . . . ....

          11. 5.11. . . . . . . . . . ....

          12. 5.12. . . . . . . . . . ....

          13. 5.13. . . . . . . . . . ....

          14. 5.14. . . . . . . . . . ....

          15. 5.15. . . . . . . . . . ....

          16. 5.16. . . . . . . . . . ....

          17. 5.17. . . . . . . . . . ....

          18. 5.18. . . . . . . . . . ....

          19. 5.19. . . . . . . . . . ....

        6. 6. PROCEDURE

          1. 6.1. Sample preparation

            1. 6.1.1. Isolation of caseins

            2. 6.1.2. Plasmin cleavage of β-caseins to intensify γ-caseins

          2. 6.2. Preparation of the urea containing polyacrylamide gels

          3. 6.3. lsoelectric focusing

            2. Conditions for isoelectric focusing:

              1. 6.3.1. Gel size 265 × 125 × 0,25 mm

              2. 6.3.2. Example of a voltage programme for an automatic electrophoresis device...

          4. 6.4. Protein staining

            1. 6.4.1. Protein fixation

            2. 6.4.2. Washing and staining the gel plate

            3. 6.4.3. Destaining the gel plate

        7. 7. EVALUATION

          2. 7.1. Visual estimation

          3. 7.2. Densitometric estimation

        8. 8. REFERENCES

    10. ANNEX X

      (Article 7)

      1. REFERENCE METHOD FOR DETECTING COLIFORMS IN BUTTER, SKIMMED-MILK POWDER, CASEIN...

        1. 1. PREPARATION OF SAMPLES

        2. 2. PROCEDURE

        3. 3. RESULTS

    11. ANNEX XI

      (Article 8)

      1. DETERMINATION OF LACTOSE IN COMPOUND FEEDINGSTUFFS

        1. 1. SCOPE AND FIELD OF APPLICATION

        2. 2. REFERENCE

        3. 3. DEFINITION

        4. 4. PRINCIPLE

        5. 5. REAGENTS

          1. 5.1. General

          2. 5.2. Lactose

          3. 5.3. Concentrated Biggs/Szijarto solution

          4. 5.4. Diluted Biggs/Szijarto solution

          5. 5.5. Preparation of HPLC grade water

        6. 6. APPARATUS

          2. 6.1. HPLC ion exchange resin column

          3. 6.2. Guard column

          4. 6.3. Column oven

          5. 6.4. HPLC pump

          6. 6.5. HPLC injection device

          7. 6.6. HPLC detector

          8. 6.7. Integrator

          9. 6.8. Water purification unit

          10. 6.9. Solvent filtration unit

          11. 6.10. Analytical balance

          12. 6.11. Water bath

          13. 6.12. Centrifuge

          14. 6.13. Volumetric flask 50 mL

          15. 6.14. Volumetric flask 100 mL

          16. 6.15. Graduated pipette

        7. 7. SAMPLING

        8. 8. PREPARATION OF LACTOSE STANDARD SOLUTION

          1. 8.1. Standard 1

          2. 8.2. Standard 2

        9. 9. PREPARATION OF THE TEST SAMPLE

          1. 9.1. Reconstitution of the sample

          2. 9.2. Sample treatment

        10. 10. HPLC DETERMINATION

          1. 10.1. Preliminary preparation of HPLC

            1. 10.1.1. Installation of the column and pre-column

            2. 10.1.2. Detector and initial flow

            3. 10.1.3. Column oven and final flow-rate

            4. 10.1.4. Integration

            5. 10.1.5. Guard column test

          2. 10.2. Running standards

          3. 10.3. Running samples

        11. 11. CALCULATION AND EXPRESSION OF THE RESULTS

          1. 11.1. Calibration

          2. 11.2. Samples

        12. 12. PRECISION

          2. 12.1. Repeatability

          3. 12.2. Reproducibility

        13. 13. REFERENCES

    12. ANNEX XII

      (Article 9)

      1. DETECTION OF RENNET WHEY IN SKIMMED-MILK POWDER FOR PUBLIC STORAGE...

        1. 1. SCOPE AND FIELD OF APPLICATION

        2. 2. REFERENCE

        3. 3. DEFINITION

        4. 4. PRINCIPLE

        5. 5. REAGENTS

          2. 5.1. Trichloroacetic acid solution

          3. 5.2. Eluent solution, pH 6,0

          4. 5.3. Flushing solvent

          5. 5.4. Standard samples

            1. 5.4.1. . . . . . . . . . ....

            2. 5.4.2. . . . . . . . . . ....

        6. 6. APPARATUS

          1. 6.1. . . . . . . . . . ....

          2. 6.2. . . . . . . . . . ....

          3. 6.3. . . . . . . . . . ....

          4. 6.4. . . . . . . . . . ....

          5. 6.5. . . . . . . . . . ....

          6. 6.6. . . . . . . . . . ....

          7. 6.7. . . . . . . . . . ....

          8. 6.8. . . . . . . . . . ....

          9. 6.9. . . . . . . . . . ....

          10. 6.10. . . . . . . . . . ....

          11. 6.11. . . . . . . . . . ....

            2. 6.11.1. . . . . . . . . . ....

            3. 6.11.2. . . . . . . . . . ....

            4. 6.11.3. . . . . . . . . . ....

            5. 6.11.4. . . . . . . . . . ....

            6. 6.11.5. . . . . . . . . . ....

            7. 6.11.6. . . . . . . . . . ....

        7. 7. SAMPLING

          1. 7.1. . . . . . . . . . ....

          2. 7.2. . . . . . . . . . ....

        8. 8. PROCEDURE

          1. 8.1. Preparation of the test sample

          2. 8.2. Test portion

          3. 8.3. Removal of fat and proteins

            1. 8.3.1. . . . . . . . . . ....

            2. 8.3.2. . . . . . . . . . ....

            3. 8.3.3. . . . . . . . . . ....

          4. 8.4. Chromatographic determination

            1. 8.4.1. . . . . . . . . . ....

            2. 8.4.2. . . . . . . . . . ....

          5. 8.5. Calibration

            1. 8.5.1. . . . . . . . . . ....

            2. 8.5.2. . . . . . . . . . ....

            3. 8.5.3. . . . . . . . . . ....

        9. 9. EXPRESSION OF RESULTS

          1. 9.1. Method of calculation and formulae

            1. 9.1.1. Calculation of the response factors R:

            2. 9.1.2. Calculation of the relative area of the peaks in the...

            3. 9.1.3. Calculation of the relative retention time of peak III in...

            4. 9.1.4. Experiments have shown that there is a linear relation between...

          2. 9.2. Calculation of the percentage of rennet whey powder in the...

          3. 9.3. Accuracy of the procedure

            1. 9.3.1. Repeatability

            2. 9.3.2. Reproducibility

          4. 9.4. Interpretation

            1. 9.4.1. . . . . . . . . . ....

            2. 9.4.2. . . . . . . . . . ....

            3. 9.4.3. . . . . . . . . . ....

              2. 9.4.3.1. . . . . . . . . . ....

              3. 9.4.3.2. . . . . . . . . . ....

              4. Skimmed-milk powder

              5. Skimmed-milk powder

    13. ANNEX XIII

      (Article 9)

      1. DETERMINING RENNET WHEY SOLIDS IN SKIMMED-MILK POWDER AND THE MIXTURES...

        1. 1. PURPOSE: DETECTING THE ADDITION OF RENNET WHEY SOLIDS TO:

        2. 2. REFERENCES: INTERNATIONAL STANDARD ISO 707

        3. 3. DEFINITION

        4. 4. PRINCIPLE

        5. 5. REAGENTS

          2. 5.1. Trichloroacetic acid solution

          3. 5.2. Eluents A and B

          4. 5.3. Conservation of the column

          5. 5.4. Standard samples

            1. 5.4.1. . . . . . . . . . ....

            2. 5.4.2. . . . . . . . . . ....

            3. 5.4.3. . . . . . . . . . ....

        6. 6. APPARATUS

          2. 6.1. . . . . . . . . . ....

          3. 6.2. . . . . . . . . . ....

          4. 6.3. . . . . . . . . . ....

          5. 6.4. . . . . . . . . . ....

          6. 6.5. . . . . . . . . . ....

          7. 6.6. . . . . . . . . . ....

          8. 6.7. . . . . . . . . . ....

          9. 6.8. . . . . . . . . . ....

          10. 6.9. . . . . . . . . . ....

          11. 6.10. . . . . . . . . . ....

          12. 6.11. . . . . . . . . . ....

            2. 6.11.1. . . . . . . . . . ....

            3. 6.11.2. . . . . . . . . . ....

            4. 6.11.3. . . . . . . . . . ....

            5. 6.11.4. . . . . . . . . . ....

            6. 6.11.5. . . . . . . . . . ....

            7. 6.11.6. . . . . . . . . . ....

        7. 7. SAMPLING

          1. 7.1. . . . . . . . . . ....

          2. 7.2. . . . . . . . . . ....

        8. 8. PROCEDURE

          1. 8.1. Preparation of the test sample

          2. 8.2. Test portion

          3. 8.3. Removal of fat and proteins

            1. 8.3.1. . . . . . . . . . ....

            2. 8.3.2. . . . . . . . . . ....

            3. 8.3.3. . . . . . . . . . ....

          4. 8.4. Chromatographic determination

            1. 8.4.1. . . . . . . . . . ....

            2. 8.4.2. . . . . . . . . . ....

            3. 8.4.3. . . . . . . . . . ....

            4. 8.4.4. . . . . . . . . . ....

          5. 8.5. Calibration

            1. 8.5.1. . . . . . . . . . ....

            2. 8.5.2. . . . . . . . . . ....

            3. 8.5.3. . . . . . . . . . ....

        9. 9. EXPRESSION OF RESULTS

          1. 9.1. Method of calculation and formulae

            1. 9.1.1. Calculation of the response factor R:

          2. 9.2. Calculation of the percentage of rennet whey powder in the...

          3. 9.3. Accuracy of the procedure

            1. 9.3.1. Repeatability

            2. 9.3.2. Reproducibility

            3. 9.3.3. Linearity

          4. 9.4. Interpretation

            2. Table 1 Ni –4,6 standard

    14. ANNEX XIV

      (Article 10)

      1. SKIMMED-MILK POWDER: QUANTITATIVE DETERMINATION OF PHOSPHATIDYLSERINE AND PHOSPHATIDYLETHANOLAMINE

        2. 1. PURPOSE AND FIELD OF APPLICATION

        3. 2. DEFINITION

        4. 3. PRINCIPLE OF THE METHOD

        5. 4. REAGENTS

          2. 4.1. Standard material: PEDP, at least 99 % pure

          3. 4.2. Reagents for standard sample and test sample preparation

            1. 4.2.1. . . . . . . . . . ....

            2. 4.2.2. . . . . . . . . . ....

            3. 4.2.3. . . . . . . . . . ....

          4. 4.3. Reagents for o-phthaldialdehyde derivatisation

            1. 4.3.1. . . . . . . . . . ....

            2. 4.3.2. . . . . . . . . . ....

            3. 4.3.3. . . . . . . . . . ....

            4. 4.3.4. . . . . . . . . . ....

          5. 4.4. HPLC elution solvents

            1. 4.4.1. . . . . . . . . . ....

            2. 4.4.2. . . . . . . . . . ....

            3. 4.4.3. . . . . . . . . . ....

            4. 4.4.4. . . . . . . . . . ....

            5. 4.4.5. . . . . . . . . . ....

            6. 4.4.6. . . . . . . . . . ....

            7. 4.4.7. . . . . . . . . . ....

        6. 5. APPARATUS

          1. 5.1. . . . . . . . . . ....

          2. 5.2. . . . . . . . . . ....

          3. 5.3. . . . . . . . . . ....

          4. 5.4. . . . . . . . . . ....

          5. 5.5. . . . . . . . . . ....

          6. 5.6. . . . . . . . . . ....

          7. 5.7. . . . . . . . . . ....

          8. 5.8. . . . . . . . . . ....

          9. 5.9. . . . . . . . . . ....

          10. 5.10. . . . . . . . . . ....

          11. 5.11. . . . . . . . . . ....

          12. 5.12. . . . . . . . . . ....

          13. 5.13. . . . . . . . . . ....

            2. 5.13.1. . . . . . . . . . ....

            3. 5.13.2. . . . . . . . . . ....

            4. 5.13.3. . . . . . . . . . ....

            5. 5.13.4. . . . . . . . . . ....

            6. 5.13.5. . . . . . . . . . ....

            7. 5.13.6. . . . . . . . . . ....

        7. 6. SAMPLING

        8. 7. PROCEDURE

          1. 7.1. Preparation of the internal standard solution

            1. 7.1.1. . . . . . . . . . ....

            2. 7.1.2. . . . . . . . . . ....

          2. 7.2. Preparation of the test sample solution

            1. 7.2.1. . . . . . . . . . ....

            2. 7.2.2. . . . . . . . . . ....

            3. 7.2.3. . . . . . . . . . ....

          3. 7.3. Preparation of the external standard solution

            1. 7.3.1. . . . . . . . . . ....

            2. 7.3.2. . . . . . . . . . ....

          4. 7.4. Preparation of the derivatising reagent

          5. 7.5. Determination by HPLC

            1. 7.5.1. Elution solvents (4.4)

            2. 7.5.2. Suggested eluting gradient:

            3. 7.5.3. . . . . . . . . . ....

            4. 7.5.4. Column equilibration

            5. 7.5.5. . . . . . . . . . ....

            6. 7.5.6. Perform the sequence of the chromatographic analyses keeping constant the...

          6. 7.6. Integration mode

            1. 7.6.1. PEDP peak

            2. 7.6.2. Tryptamine peak

            3. 7.6.3. PS and PE peaks groups

        9. 8. CALCULATION AND EXPRESSION OF RESULTS

        10. 9. ACCURACY OF THE METHOD

          2. 9.1. Repeatability

          3. 9.2. Reproducibility

        11. 10. REFERENCES

          1. 10.1. . . . . . . . . . ....

    15. ANNEX XV

      (Article 11)

      1. DETECTION OF ANTIMICROBIAL RESIDUES IN SKIMMED MILK POWDER

        2. Positive results are to be interpreted as follows:

    16. ANNEX XVI

      (Article 12)

      1. QUANTITATIVE DETERMINATION OF SKIMMED-MILK POWDER IN COMPOUND FEEDINGSTUFFS BY ENZYMATIC...

        1. 1. PURPOSE

        2. 2. SCOPE

        3. 3. PRINCIPLE OF THE METHOD

          1. 3.1. . . . . . . . . . ....

          2. 3.2. . . . . . . . . . ....

          3. 3.3. . . . . . . . . . ....

        4. 4. REAGENTS

          2. 4.1. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

          4. 4.3. . . . . . . . . . ....

          5. 4.4. . . . . . . . . . ....

          6. 4.5. . . . . . . . . . ....

          7. 4.6. . . . . . . . . . ....

        5. 5. APPARATUS

          2. 5.1. . . . . . . . . . ....

          3. 5.2. . . . . . . . . . ....

          4. 5.3. . . . . . . . . . ....

          5. 5.4. . . . . . . . . . ....

          6. 5.5. . . . . . . . . . ....

          7. 5.6. . . . . . . . . . ....

          8. 5.7. . . . . . . . . . ....

          9. 5.8. . . . . . . . . . ....

          10. 5.9. . . . . . . . . . ....

          11. 5.10. . . . . . . . . . ....

          12. 5.11. . . . . . . . . . ....

          13. 5.12. . . . . . . . . . ....

        6. 6. PROCEDURE

          1. 6.1. Preparation of the sample

          2. 6.2. Dissolving of milk powder and separation of the insoluble residue...

            1. 6.2.1. . . . . . . . . . ....

            2. 6.2.2. . . . . . . . . . ....

            3. 6.2.3. . . . . . . . . . ....

            4. 6.2.4. . . . . . . . . . ....

          3. 6.3. Coagulation of casein with the enzymes of rennet

            1. 6.3.1. . . . . . . . . . ....

            2. 6.3.2. . . . . . . . . . ....

            3. 6.3.3. . . . . . . . . . ....

            4. 6.3.4. . . . . . . . . . ....

          4. 6.4. Determination of casein nitrogen

            1. 6.4.1. . . . . . . . . . ....

        7. 7. BLANK TEST

          1. 7.1. . . . . . . . . . ....

          2. 7.2. . . . . . . . . . ....

        8. 8. EXPRESSION OF RESULTS

          1. 8.1. . . . . . . . . . ....

        9. 9. ACCURACY OF THE METHOD

          1. 9.1. Repeatability

          2. 9.2. Reproducibility

        10. 10. OBSERVATIONS

          1. 10.1. . . . . . . . . . ....

          2. 10.2. . . . . . . . . . ....

          3. 10.3. . . . . . . . . . ....

          4. 10.4. . . . . . . . . . ....

    17. ANNEX XVII

      (Article 13)

      1. DETECTION OF STARCH IN SKIMMED-MILK POWDER, DENATURED MILK POWDER AND...

        1. 1. SCOPE

        2. 2. PRINCIPLE

        3. 3. REAGENTS

          1. 3.1. . . . . . . . . . ....

        4. 4. APPARATUS

          1. 4.1. . . . . . . . . . ....

          2. 4.2. . . . . . . . . . ....

          3. 4.3. . . . . . . . . . ....

        5. 5. PROCEDURE

        6. 6. EXPRESSION OF RESULTS

        7. 7. REMARKS

    18. ANNEX XVIII

      (Article 14)

      1. DETERMINATION OF MOISTURE CONTENT IN DRIED CREAM

        1. 1. SCOPE

        2. 2. TERMS AND DEFINITIONS

        3. 3. PRINCIPLE

        4. 4. APPARATUS

          2. 4.1. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

          4. 4.3. . . . . . . . . . ....

          5. 4.4. . . . . . . . . . ....

          6. 4.5. . . . . . . . . . ....

        5. 5. SAMPLING

        6. 6. PREPARATION OF TEST SAMPLE

        7. 7. PROCEDURE

          1. 7.1. Preparation of the dish

            1. 7.1.1. . . . . . . . . . ....

            2. 7.1.2. . . . . . . . . . ....

          2. 7.2. Test portion

          3. 7.3. Determination

            1. 7.3.1. . . . . . . . . . ....

            2. 7.3.2. . . . . . . . . . ....

            3. 7.3.3. . . . . . . . . . ....

            4. 7.3.4. . . . . . . . . . ....

        8. 8. CALCULATION AND EXPRESSION OF RESULTS

          1. 8.1. Calculation

        9. 9. PRECISION

          2. 9.1. Repeatability

          3. 9.2. Reproducibility

        10. 10. TEST REPORT

    19. ANNEX XIX

      (Article 15)

      1. DETERMINATION OF MOISTURE IN ACID BUTTERMILK POWDER

        1. 1. SCOPE

        2. 2. PRINCIPLE

        3. 3. APPARATUS

          1. 3.1. . . . . . . . . . ....

          2. 3.2. . . . . . . . . . ....

          3. 3.3. . . . . . . . . . ....

          4. 3.4. . . . . . . . . . ....

          5. 3.5. . . . . . . . . . ....

        4. 4. PROCEDURE

        5. 5. CALCULATION

        6. 6. PRECISION

          1. 6.1. Repeatability limit

          2. 6.2. Reproducibility limit

          3. 6.3. Source of precision data

    20. ANNEX XX

      (Article 16)

      1. REFERENCE METHOD FOR THE DETERMINATION OF MILK FAT PURITY BY...

        1. 1. SCOPE AND FIELD OF APPLICATION

        2. 2. DEFINITION

        3. 3. PRINCIPLE OF THE METHOD

        4. 4. REAGENTS

          2. 4.1. . . . . . . . . . ....

          3. 4.2. . . . . . . . . . ....

            2. 4.2.1. . . . . . . . . . ....

            3. 4.2.2. . . . . . . . . . ....

          4. 4.3. . . . . . . . . . ....

          5. 4.4. . . . . . . . . . ....

          6. 4.5. . . . . . . . . . ....

          7. 4.6. . . . . . . . . . ....

          8. 4.7. . . . . . . . . . ....

        5. 5. APPARATUS

          2. 5.1. High-temperature gas chromatograph

          3. 5.2. Chromatography column

            1. 5.2.1. Packed column

            2. 5.2.2. Capillary column

          4. 5.3. . . . . . . . . . ....

          5. 5.4. . . . . . . . . . ....

          6. 5.5. . . . . . . . . . ....

          7. 5.6. . . . . . . . . . ....

          8. 5.7. . . . . . . . . . ....

          9. 5.8. . . . . . . . . . ....

          10. 5.9. . . . . . . . . . ....

          11. 5.10. . . . . . . . . . ....

          12. 5.11. . . . . . . . . . ....

          13. 5.12. . . . . . . . . . ....

          14. 5.13. . . . . . . . . . ....

          15. 5.14. . . . . . . . . . ....

          16. 5.15. . . . . . . . . . ....

          17. 5.16. . . . . . . . . . ....

        6. 6. SAMPLING

        7. 7. PROCEDURE

          1. 7.1. Preparation of test samples

            2. 7.1.1. Isolation from butter or butteroil

            3. 7.1.2. Extraction according to the Röse–Gottlieb gravimetric method

            4. 7.1.3. Extraction from milk using silica gel columns

          2. 7.2. Preparation of sample solution

          3. 7.3. Chromatographic triglyceride determination

            1. 7.3.1. Baseline drift

            2. 7.3.2. Injection technique

              1. 7.3.2.1. Packed column

              2. 7.3.2.2. Capillary column

            3. 7.3.3. Calibration

              1. 7.3.3.1. General

              2. 7.3.3.2. Commercial milk fat standard

              3. 7.3.3.3. Laboratory milk fat standard

            4. 7.3.4. Chromatographic conditions

              2. 7.3.4.1. Packed column

              3. 7.3.4.2. Capillary column

        8. 8. INTEGRATION, EVALUATION AND CONTROL OF THE ANALYTICAL PERFORMANCE

        9. 9. CALCULATION AND EXPRESSION OF RESULTS

          1. 9.1. Triglyceride composition

            1. 9.1.1. Calculation

            2. 9.1.2. Expression of test results

          2. 9.2. S-values

            1. 9.2.1. Calculation

              1. 9.2.1.1. . . . . . . . . . ....

              2. 9.2.1.2. . . . . . . . . . ....

              3. 9.2.1.3. . . . . . . . . . ....

              4. 9.2.1.4. . . . . . . . . . ....

              5. 9.2.1.5. . . . . . . . . . ....

              6. 9.2.1.6. . . . . . . . . . ....

            2. 9.2.2. Expression of test results

          3. 9.3. Detection of foreign fat

        10. 10. PRECISION

          1. 10.1. Interlaboratory test

          2. 10.2. Repeatability

          3. 10.3. Reproducibility

        11. 11. UNCERTAINTY OF MEASUREMENT

        12. 12. TEST REPORT

      2. ANNEX A

        (normative)

        1. PREPARATION OF THE PACKED COLUMN

          1. A.1 REAGENTS AND APPARATUS

            1. A.1.1 . . . . . . . . . ....

            2. A.1.2 . . . . . . . . . ....

            3. A.1.3 . . . . . . . . . ....

            4. A.1.4 . . . . . . . . . ....

            5. A.1.5 . . . . . . . . . ....

            6. A.1.6 . . . . . . . . . ....

              2. A.1.6.1 . . . . . . . . . ....

              3. A.1.6.2 . . . . . . . . . ....

              4. A.1.6.3 . . . . . . . . . ....

              5. A.1.6.4 . . . . . . . . . ....

              6. A.1.6.5 . . . . . . . . . ....

              7. A.1.6.6 . . . . . . . . . ....

              8. A.1.6.7 . . . . . . . . . ....

          2. A.2 SILANIZATION (DEACTIVATION OF THE GLASS SURFACE)

          3. A.3 FILLING

          4. A.4 CONDITIONING

      3. ANNEX B

        (informative)

        1. QUANTIFICATION OF THE FOREIGN FAT CONTENT

          1. B.1 GENERAL

          2. B.2 CALCULATION

          3. B.3 EXPRESSION OF TEST RESULTS

          4. Bibliography

    21. ANNEX XXI

      (Article 18)

      1. PROCEDURE APPLICABLE WHEN THE RESULTS OF AN ANALYSIS ARE DISPUTED...

        1. 1. . . . . . . . . . ....

        2. 2. . . . . . . . . . ....

        3. 3. . . . . . . . . . ....

        4. 4. . . . . . . . . . ....

    22. ANNEX XXII

      CORRELATION TABLE

      1. . . . . . . . . . ....

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