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Commission Regulation (EC) No 152/2009 of 27 January 2009 laying down the methods of sampling and analysis for the official control of feed (Text with EEA relevance)
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Article 1.Sampling for the official control of feed, in particular as...
Article 2.Preparation of samples for analysis and expression of results shall...
Article 3.Analysis for the official control of feed shall be carried...
Article 4.The energy value of compound poultry feed shall be calculated...
Article 5.The methods of analysis to control illegal presence of no...
Article 6.Directives 71/250/EEC, 71/393/EEC, 72/199/EEC, 73/46/EEC, 76/371/EEC, 76/372/EEC, 78/633/EEC, 81/715/EEC, 84/425/EEC,...
Article 7.This Regulation shall enter into force on the twentieth day...
5. QUANTITATIVE REQUIREMENTS AS REGARDS NUMBER OF INCREMENTAL SAMPLES
8. METHOD OF SAMPLING FOR VERY LARGE LOTS OR LOTS STORED...
9. INSTRUCTIONS FOR TAKING, PREPARING AND PACKAGING THE SAMPLES
METHODS OF ANALYSIS TO CONTROL THE COMPOSITION OF FEED MATERIALS AND COMPOUND FEED
3.1. Crusher of non-moisture-absorbing material which is easy to clean, allows...
3.3. Dry containers of non-corrodible metal or of glass with lids...
3.4. Electrically heated isothermal oven (± 2 oC) properly ventilated and...
3.5. Adjustable electrically heated vacuum oven fitted with an oil pump...
3.6. Desiccator with a thick perforated metal or porcelain plate, containing...
B. DETERMINATION OF MOISTURE IN ANIMAL AND VEGETABLE FATS AND OILS...
C. DETERMINATION OF THE CONTENT OF CRUDE PROTEIN
3.2. Catalyst: copper (II) oxide CuO or copper (II) sulphate pentahydrate,...
3.5. Sulphuric acid, standard volumetric solution, c(H2SO4) = 0,25 mol/l.
3.6. Sulphuric acid, standard volumetric solution, c(H2SO4) = 0,1 mol/l.
3.7. Sulphuric acid, standard volumetric solution, c(H2SO4) = 0,05 mol/l.
3.8. Methyl red indicator; dissolve 300 mg of methyl red in...
3.9. Sodium hydroxide solution (Technical grade may be used) β =...
3.10. Sodium hydroxide, standard volumetric solution c(NaOH) = 0,25 mol/l.
3.11. Sodium hydroxide, standard volumetric solution c(NaOH) = 0,1 mol/l.
3.12. Granulated pumice stone, washed in hydrochloric acid and ignited.
3.16. Methyl red indicator solution: dissolve 100 mg methyl red in...
3.17. Bromocresol green solution: dissolve 100 mg bromocresol green in 100...
3.19. Hydrochloric acid standard volumetric solution c(HCl) = 0,1 mol/l.
E. DETERMINATION OF VOLATILE NITROGENOUS BASES
F. DETERMINATION OF AMINO ACIDS (EXCEPT TRYPTOPHANE)
3.13. Norleucine, or other compound suitable for use as internal standard....
3.16.1. Standard substances listed under paragraph 1. Pure compounds containing no...
3.21. Extraction mixture, c = 0,1 mol HCl/l containing 2 %...
3.25. Elution buffers, prepared according to conditions for the analyser used...
3.26. Ninhydrin reagent, prepared according to conditions for the analyser used...
3.27. Standard solutions of amino acids. These solutions shall be stored...
3.27.2. Stock standard solution of cysteic acid and methionine sulphone, c...
3.27.3. Stock standard solution of the internal standard e.g. norleucine, c...
3.27.4. Calibration solution of standard amino acids for use with hydrolysates,...
3.27.5. Calibration solution of standard amino acids for use with hydrolysates...
G. DETERMINATION OF TRYPTOPHAN
3.1. Double distilled water or water of equivalent quality must be...
3.2. Standard substance: tryptophan (purity/content ≥ 99 %) dried under vacuum...
3.3. Internal standard substance: α-methyl-tryptophan (purity/content ≥ 99 %), dried under...
3.4. Barium hydroxide octa-hydrate (care shall be taken not to expose...
3.15. Concentrated solution of tryptophan (3.2), c = 2,5 μmol/ml:
3.16. Concentrated internal standard solution, c = 2,5 μmol/ml:
3.17. Calibration standard solution of tryptophan and internal standard:
3.21. Solution of 1 g 1,1,1-trichloro-2-methyl-2-propanol (3.19) in 100 ml methanol...
I. DETERMINATION OF CRUDE FIBRE
4.1. Heating unit for digestion with sulphuric acid and potassium hydroxide...
4.2. Glass filter crucible with fused sintered glass filter plate pore...
4.3. Cylinder of at least 270 ml with a reflux condenser,...
4.6. Extraction unit consisting of a support plate for the filter...
4.7. Connecting rings to assemble the heating unit (4.1), crucible (4.2)...
3.1. Ethanol solution 40 % (v/v) density: 0,948 g/ml at 20...
3.3. Carrez solution II: dissolve in water 10,6 g of potassium...
3.8.1. Copper sulphate solution: dissolve 25 g of copper sulphate, Cu...
3.8.2. Citric acid solution: dissolve 50 g of citric acid, C6H8O7·H2O...
3.8.3. Sodium carbonate solution: dissolve 143,8 g of anhydrous sodium carbonate...
3.13. Granulated pumice stone boiled in hydrochloric acid, washed in water...
8.2. The difference between the content of total sugars after inversion,...
8.3. In order to determine the content of reducing sugars, excluding...
8.3.1. For an approximate calculation, multiply by 0,675 the lactose content...
8.3.2. For an accurate calculation of reducing sugars, excluding lactose, the...
3.1. Suspension of Saccharomyces cerevisiae: suspend 25 g of fresh yeast...
3.3. Carrez solution II: dissolve in water 10,6 g of potassium...
3.4.1. Copper sulphate solution: dissolve 25 g of copper sulphate Cu...
3.4.2. Citric acid solution: dissolve 50 g of citric acid C6H8O7·H2O...
3.4.3. Sodium carbonate solution: dissolve 143,8 g of anhydrous sodium carbonate...
3.5. Granulated pumice stone boiled in hydrochloric acid, washed in water...
N. DETERMINATION OF ASH WHICH IS INSOLUBLE IN HYDROCHLORIC ACID
O. DETERMINATION OF CARBONATES
P. DETERMINATION OF TOTAL PHOSPHORUS
3.2. Hydrochloric acid, ρ20 = 1,1 g/ml (approx 6 mol/litre).
3.6. Molybdovanadate reagent: mix 200 ml of ammonium heptamolybdate solution (3.6.1),...
3.6.1. Ammonium heptamolybdate solution: dissolve in hot water 100 g of...
3.6.2. Ammonium monovanadate solution: dissolve 2,35 g of ammonium monovanadate NH4VO3...
3.7. Standard solution of 1 mg phosphorus per ml: dissolve 4,387...
METHODS OF ANALYSIS TO CONTROL THE LEVEL OF AUTHORISED ADDITIVES IN FEED
3.6.1. Sodium sulphide solution, c = 0,5 mol/l in glycerol, ß...
3.7. Phenolphthalein solution, c = 2 g/100 ml in ethanol (3.1)...
3.9. Mobile phase for HPLC: mixture of methanol (3.3) and water,...
3.11. All-trans-vitamin A acetate, extra pure, of certified activity, e.g. 2,8...
3.11.1. Stock solution of all-trans-vitamin A acetate: Weigh to the nearest...
3.12. All-trans-vitamin A palmitate, extra pure, of certified activity, e.g. 1,8...
3.12.1. Stock solution of all-trans-vitamin A palmitate: Weigh to the nearest...
3.13. 2,6-Di-tert-butyl-4-methylphenol (BHT) (see 7.5 observations)
4.2.1. Flat bottom or conical flasks, 500 ml, with ground-glass socket...
4.2.2. Graduated flasks with ground-glass stoppers, narrow-necked, 10, 25, 100 and...
4.2.3. Separating funnels, conical, 1 000 ml, with ground-glass stoppers
4.2.4. Pear shaped flasks, 250 ml, with ground-glass sockets
4.3. Allihn condenser, jacket length 300 mm, with ground-glass joint, with...
4.4. Pleated filter paper for phase separation, diameter 185 mm (e.g....
4.5.1. Liquid chromatographic column, 250 mm x 4 mm, C18, 5...
4.5.2. UV or fluorescence detector, with variable wavelength adjustment
4.8.1. Glass cylinder of 1 l capacity fitted with a ground...
4.8.2. Ground glass insert equipped with a side-arm and an adjustable...
7.1. For samples with low vitamin A concentration it may be...
7.2. The weight of the sample taken for the analysis shall...
7.3. If phase separation does not occur add approximately 10 ml...
7.4. With cod-liver oil and other pure fats the saponification time...
7.6. Using a normal phase-column the separation of retinol isomers is...
7.7. Approximately 150 mg ascorbic acid can be used instead of...
7.8. Approximately 50 mg EDTA can be used instead of sodium...
7.9. In cases of analysis of vitamin A in milk replacers,...
3.6.1. Sodium sulphide solution, c = 0,5 mol/l in glycerol, β...
3.7. Phenolphthalein solution, c = 2 g/100 ml in ethanol (3.1)....
3.8. Mobile phase for HPLC: mixture of methanol (3.3) and water,...
3.10. DL-α-tocopherol acetate, extra pure, of certified activity.
3.10.1. Stock solution of DL-α-tocopherol acetate: Weigh to the nearest 0,1...
3.11.1. Stock solution of DL-α-tocopherol: Weigh to the nearest 0,1 mg,...
3.12. 2,6-Di-tert-butyl-4-methylphenol (BHT) (see 7.5 observations).
4.2.1. Flat bottom or conical flasks, 500 ml, with ground-glass socket....
4.2.2. Graduated flasks with ground-glass stoppers, narrow-necked, 10, 25, 100 and...
4.2.3. Separating funnels, conical, 1 000 ml, with ground-glass stoppers.
4.2.4. Pear shaped flasks, 250 ml, with ground-glass sockets.
4.3. Allihn condenser, jacket length 300 mm, with ground-glass joint, with...
4.4. Pleated filter paper for phase separation, diameter 185 mm (e.g....
4.5.1. Liquid chromatographic column, 250 mm × 4 mm, C18, 5...
4.5.2. Fluorescence or UV detector, with variable wavelength adjustment.
4.8.1. Glass cylinder of 1 l capacity fitted with a ground...
4.8.2. Ground glass insert equipped with a side-arm and an adjustable...
7.1. For samples with low vitamin E concentration it may be...
7.2. The weight of the sample taken for the analysis shall...
7.3. If phase separation does not occur add approximately 10 ml...
7.4. After the spectrophotometric measurement of the DL-α-tocopherol acetate or DL-α-tocopherol...
7.6. Using a normal phase-column the separation of α-, β-, γ-...
7.7. Approximately 150 mg ascorbic acid can be used instead of...
7.8. Approximately 50 mg EDTA can be used instead of sodium...
7.9. Vitamin E acetate hydrolyses very fast under alkaline conditions and...
C. DETERMINATION OF THE TRACE ELEMENTS IRON, COPPER, MANGANESE AND ZINC...
3.6. Hydrogen peroxide (approximately 100 volumes of oxygen (30 % by...
3.7. Standard iron solution (1 000 μg Fe/ml) prepared as follows...
3.7.1. Working standard iron solution (100 μg Fe/ml) prepared by diluting...
3.8. Standard copper solution (1 000 μg Cu/ml) prepared as follows...
3.8.1. Working standard copper solution (10 μg Cu/ml) prepared by diluting...
3.9. Standard manganese solution (1 000 μg Mn/ml) prepared as follows...
3.9.1. Working standard manganese solution (10 μg Mn/ml) prepared by diluting...
3.10. Standard zinc solution (1 000 μg Zn/ml) prepared as follows...
3.10.1. Working standard zinc solution (10 μg Zn/ml) prepared by diluting...
3.11. Lanthanum chloride solution: dissolve 12 g of lanthanum oxide in...
D. DETERMINATION OF HALOFUGINONE
E. DETERMINATION OF ROBENIDINE
F. DETERMINATION OF DICLAZURIL
G. DETERMINATION OF LASALOCID SODIUM
METHODS OF ANALYSIS TO CONTROL UNDESIRABLE SUBSTANCES IN FEED
A. DETERMINATION OF FREE AND TOTAL GOSSYPOL
3.1. Propan-2-ol-hexane mixture: mix 60 parts by volume of propan-2-ol with...
3.2. Solvent A: Place in a 1 litre graduated flask approximately...
3.3. Solvent B: Pipette 2 ml of 3-aminopropan-1-ol and 10 ml...
3.4. Aniline: If the optical density in the blank test exceeds...
3.5. Standard gossypol solution A: Place 27,9 mg of gossypol acetate...
3.6. Standard gossypol solution B: Place 27,9 mg of gossypol acetate...
B. DETERMINATION OF THE LEVELS OF DIOXINS (PCDD/PCDF) AND PCBs
CHAPTER I Methods of sampling and interpretation of analytical results
CHAPTER II Sample preparation and requirements for methods of analysis used in...
3. Quality assurance requirements
3.1. Measures shall be taken to avoid cross-contamination at each stage...
3.2. The samples shall be stored and transported in glass, aluminum,...
3.3. The sample storage and transportation shall be performed in a...
3.4. Insofar as relevant, each laboratory sample shall be finely grinded...
3.5. Control of reagents, glassware and equipment for possible influence of...
3.6. A blank analysis shall be performed by carrying out the...
3.7. For bioanalytical methods, all glassware and solvents used in analysis...
3.8. Sample quantity used for the extraction shall be sufficient to...
3.9. The specific sample preparation procedures used for the products under...
5. Basic requirements to be met by analytical procedure for dioxins...
5.3. High accuracy (trueness and precision, bioassay apparent recovery)
5.4. Validation in the range of maximum level and general quality...
5.7. Specific requirements for screening methods
5.7.1. Both GC-MS and bioanalytical methods may be used for screening....
5.7.2. Laboratories applying screening methods for the routine control of samples...
5.7.3. Performance verification of the screening method is required during routine...
5.7.4. Check on possible suppression of the cell response and cytotoxicity:...
5.7.6. Determination of false-compliant rates from quality control data:
5.7.7. Potential non-compliant samples from screening shall always be verified by...
5.7.8. Under validation conditions, bioanalytical methods shall provide a valid indication...
6. SPECIFIC requirements for GC-MS methods to be complied with for...
6.1. Acceptable differences between upper-bound and lower-bound WHO-TEQ results
6.2.1. Addition of 13 C-labelled 2,3,7,8-chlorine-substituted internal PCDD/F standards and of...
6.2.2. Relative response factors shall also be determined for those congeners...
6.2.3. For feed of plant origin and feed of animal origin...
6.2.4. Prior to GC-MS analysis, 1 or 2 recovery (surrogate) standard(s)...
6.2.5. Control of recovery is required. For confirmatory methods, the recoveries...
7. Specific requirements for bioanalytical methods
7.4. Performance characteristics
7.4.1. Since no internal standards can be used in bioanalytical methods,...
7.4.2. As part of the validation process, the test shall be...
7.4.3. Target compounds, possible interferences and maximum tolerable blank levels shall...
7.4.4. The percent standard deviation in the response or concentration calculated...
7.4.5. The uncorrected results of the reference sample(s) expressed in BEQ...
7.4.6. Quality control charts for procedure blanks and each type of...
7.4.7. The results from the confirmatory methods of suspected samples and...
7.4.8. Successful method performance may also be demonstrated by participation in...
7.4.9. During incidents, the cut-off values may be re-evaluated, reflecting the...
8.1.1. The analytical results shall contain the levels of the individual...
8.1.2. The report shall include the method used for extraction of...
8.1.3. The recoveries of the individual internal standards shall be made...
8.1.4. As the expanded measurement uncertainty is to be taken into...
8.1.5. The results shall be expressed in the same units and...
8.2. Bioanalytical screening methods
8.2.1. The result of the screening shall be expressed as ‘compliant’...
8.2.2. In addition, an indicative result for PCDD/Fs and/or dioxin-like PCBs...
8.2.3. Samples with a response below the reporting limit shall be...
8.2.4. For each type of sample matrix, the report shall mention...
8.2.5. The report shall mention the type of the test applied,...
8.2.6. The report shall include the method used for extraction of...
8.2.7. In case of samples suspected to be non-compliant, the report...
8.2.8. Non-compliant results shall only be reported from confirmatory analysis.
8.3. Physico-chemical screening methods
8.3.1. The result of the screening shall be expressed as ‘compliant’...
8.3.2. For each type of sample matrix, the report shall mention...
8.3.3. In addition, levels for individual PCDD/F and/or dioxin-like PCB congeners...
8.3.4. The recoveries of the individual internal standards shall be made...
8.3.6. The report shall include the method used for extraction of...
8.3.7. In case of samples suspected to be non-compliant, the report...
8.3.8. Non-compliance can only be decided after confirmatory analysis.
CHAPTER III Sample preparation and requirements for methods of analysis used in...
7.1. Suitable internal standards with physico-chemical properties comparable to analytes of...
7.3. Requirements for methods using all six isotope-labelled non-dioxin-like PCB congeners...
7.4. Requirements for methods using not all six isotope-labelled internal standards...
7.5. The recoveries of unlabelled congeners shall be checked by spiked...
9. Performance characteristics: criteria for the sum of non-dioxin-like PCBs at...
10.1. The analytical results shall contain the levels of the individual...
10.2. The report shall include the method used for the extraction...
10.3. The recoveries of the individual internal standards shall be made...
10.4. As the expanded measurement uncertainty is to be taken into...
10.5. The results shall be expressed in the same units and...
2.1.2.2.2. Grinding equipment: knife or rotor mill. If a rotor mill...
2.1.2.2.4. Conical glass separation funnel with a content of 250 ml...
2.1.2.2.5. Stereomicroscope covering at least a 6,5× to 40× final magnification...
2.1.2.2.6. Compound microscope covering at least a 100× to 400× final...
2.1.2.2.11. Filter paper: qualitative cellulose filter (pore size 4-11 μm)
2.1.3. Sampling and sample preparation
2.1.3.3. Preparation of samples other than fat or oil
2.1.3.3.1. Sample drying : samples with a moisture content > 14...
2.1.3.3.2. Sample pre-sieving : in order to collect information on possible...
2.1.3.3.3. Sub-sampling and grinding : at least 50 g of the...
2.1.3.3.4. Extraction and preparation of the sediment : a portion of...
2.1.3.3.5. Extraction and preparation of the flotate : after recovery of...
2.1.3.3.6. Preparation of raw material : a portion of at least...
METHODS OF ANALYSIS TO CONTROL ILLEGAL PRESENCE OF NO LONGER AUTHORISED ADDITIVES IN FEED
A. DETERMINATION OF METHYL BENZOQUATE
B. DETERMINATION OF OLAQUINDOX
CORRELATION TABLES REFERRED TO IN ARTICLE 6
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