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)

3. Directive 72/199/EEC

4. Directive 73/46/EEC

5. Directive 76/371/EEC

6. Directive 76/372/EEC

7. Directive 78/633/EEC

8. Directive 81/715/EEC

9. Directive 84/425/EEC

10. Directive 86/174/EEC

11. Directive 93/70/EEC

12. Directive 93/117/EC

13. Directive 98/64/EC

14. Directive 1999/27/EC

15. Directive 1999/76/EC

16. Directive 2000/45/EC

17. Directive 2002/70/EC

18. Directive 2003/126/EC

[^E0001]: Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 September 2003 on additives for use in animal nutrition (OJ L 268, 18.10.2003, p. 29).

[^E0002]: Directive 2002/32/EC of the European Parliament and of the Council of 7 May 2002 on undesirable substances in animal feed (OJ L 140, 30.5.2002, p. 10).

[^E0003]: OJ L 70, 16.3.2005, p. 1.

[^E0004]: Regulation (EC) No 767/2009 of the European Parliament and of the Council of 13 July 2009 on the placing on the market and use of feed, amending European Parliament and Council Regulation (EC) No 1831/2003 and repealing Council Directive 79/373/EEC, Commission Directive 80/511/EEC, Council Directives 82/471/EEC, 83/228/EEC, 93/74/EEC, 93/113/EC and 96/25/EC and Commission Decision 2004/217/EC (OJ L 229, 1.9.2009, p. 1).

[^E0020]: Any lumps shall be broken up (if necessary by separating them out and returning them to the sample).

[^E0021]: Except in the case of roughage/forage with low specific density.

[^E0022]: Except in the case of roughage/forage with low specific density.

[^E0023]: Regulation (EC) No 178/2002 of the European Parliament and of the Council of 28 January 2002 laying down the general principles and requirements of food law, establishing the European Food Safety Authority and laying down procedures in matters of food safety (OJ L 31, 1.2.2002, p. 1).

[^E0024]: Regulation (EU) 2019/4 of the European Parliament and of the Council of 11 December 2018 on the manufacture, placing on the market and use of medicated feed, amending Regulation (EC) No 183/2005 of the European Parliament and of the Council and repealing Council Directive 90/167/EEC (OJ L 4, 7.1.2019, p. 1).

[^E0025]: The confidence interval of 95 % can be achieved by making use of another factor such as the t-factor.

[^E0026]: The confidence interval of 95 % can be achieved by making use of another factor such as the t-factor.

[^E0027]: For the drying of cereals, flour, groats and meal, the oven must have a thermal capacity such that, when pre-set at 131 °C, it will return to that temperature in less than 45 minutes after the maximum number of test samples have been placed inside to dry simultaneously. Ventilation must be such that, when as many samples of common wheat as it can contain are dried for two hours, the results differ from those obtained after four hours of drying by less than 0,15 %.

[^E0028]: For more details on the calculation, reference is made to EN ISO 6498– Animal Feedingstuffs – Guidelines for sample preparation.

[^E0029]: N content can be determined in all feeds but conversion factor of 6,25 for the calculation of crude protein content might not be applicable for insect feed materials (lower conversion factor) and certain pet food and blood plasma proteins (higher conversion factor).

[^E0030]: The method of analysis provided for by EN 17180 is referred to as an alternative method to be used for official control purposes for the determination of amino acids in feed containing more than 10 % of amino acids.

[^E0031]: This method has not been validated through a collaborative trial for premixtures containing more than 10 % of feed additives. However, it is also applicable to these matrices with appropriate slight modifications provided that the method is then in-house validated. For additional information, consult https://ec.europa.eu/jrc/en/eurl/feed-additives/authorisation.

[^E0032]: Where the oil or fat has to undergo subsequent quality tests, replace the fragments of pumice stone by glass beads.

[^E0033]: Commission Regulation (EC) No 121/2008 of 11 February 2008 laying down the method of analysis for the determination of starch content in preparations of a kind used in animal feeding (CN code 2309 ) (OJ L 37, 12.2.2008, p. 3).

[^E0034]: The method of analysis provided for by EN 17547 is referred to as an alternative method to be used for official control purposes for the determination of vitamin A and E instead of the method described for the determination of vitamin A in part A of this Annex and for vitamin E in part B of this Annex.

[^E0035]: Conducted by the Feed Working Group of Verband Deutscher Landwirtschaftlicher Untersuchungs- und Forschungsanstalten (VDLUFA).

[^E0036]: The method of analysis provided for by EN 17547 is referred to as an alternative method to be used for official control purposes for the determination of vitamin A and E instead of the method described for the determination of vitamin A in part A of this Annex and for vitamin E in part B of this Annex.

[^E0037]: Conducted by the Feed Working Group of Verband Deutscher Landwirtschaftlicher Untersuchungs- und Forschungsanstalten (VDLUFA).

[^E0038]: This method has been validated through a collaborative trial for various feed matrices. For additional information, consult https://ec.europa.eu/jrc/en/eurl/feed-additives/authorisation.

[^E0039]: Other methods of digestion may be used provided they have been demonstrated to have similar results (such as microwave pressure digestion).

[^E0040]: Green fodder (fresh or dried) is liable to contain large amounts of vegetable silica, which may retain trace elements and must be removed. For samples of these feed, therefore, the following modified procedure must be followed. Carry out operation 5.1.1.1 as far as the filtration. Wash the filter paper containing the insoluble residue twice with boiling water and place it in a quartz or platinum crucible.

Ignite in the muffle furnace (point 4.1) at a temperature below 550 °C until all carbonaceous material has completely disappeared. Allow to cool, add a few drops of water followed by 10 to 15 ml of hydrofluoric acid (point 3.4) and evaporate to dryness at about 150 °C. If any silica remains in the residue, redissolve it in a few millilitres of hydrofluoric acid (point 3.4) and evaporate to dryness. Add five drops of sulphuric acid (point 3.5) and heat until no more white fumes are given off. After the addition of 5 ml of 6 mol/litre hydrochloric acid (point 3.2) and about 30 ml of water, heat, filter the solution into the 250 ml volumetric flask and make up to the mark with water (HCl concentration about 0,5 mol/l). Proceed then with the determination from point 5.1.2.

[^E0041]: The Analyst 108, 1983, pp. 1252 to 1256.

[^E0042]: The method may also be applicable to the determination of diclazuril in feed materials.

[^E0043]: Longer stability (up to 1 year) might be possible but it has to be confirmed by the individual laboratory.

[^E0048]: Table of TEF (= toxic equivalency factors) for PCDDs, PCDFs and dioxin-like PCBs: WHO-TEFs for human risk assessment based on the conclusions of the World Health Organization (WHO) – International Programme on Chemical Safety (IPCS) expert meeting which was held in Geneva in June 2005 (Martin van den Berg et al., The 2005 World Health Organization Re-evaluation of Human and Mammalian Toxic Equivalency Factors for Dioxins and Dioxin-like Compounds. Toxicological Sciences 93(2), 223–241 (2006)).

Abbreviations used: ‘T’ = tetra; ‘Pe’ = penta; ‘Hx’ = hexa; ‘Hp’ = hepta; ‘O’ = octa; ‘CDD’ = chlorodibenzodioxin; ‘CDF’ = chlorodibenzofuran; ‘CB’ = chlorobiphenyl.

[^E0049]: Commission Implementing Regulation (EU) 2021/808 of 22 March 2021 on the performance of analytical methods for residues of pharmacologically active substances used in food-producing animals and on the interpretation of results as well as on the methods to be used for sampling and repealing Decisions 2002/657/EC and 98/179/EC (OJ L 180, 21.5.2021, p. 84).

[^E0050]: The principles described in the ‘Guidance Document on Measurement Uncertainty for Laboratories performing PCDD/F and PCB Analysis using Isotope Dilution Mass Spectrometry’ (https://food.ec.europa.eu/system/files/2017-05/animal-feed-guidance_document_pcdd-f_pcb_en.pdf) shall be followed when applicable.

[^E0051]: The concept of ‘upper-bound’ requires using the limit of quantification for the contribution of each non-quantified congener. The concept of ‘lower-bound’ requires using zero for the contribution of each non-quantified congener. The concept of ‘medium-bound’ requires using half of the limit of quantification calculating the contribution of each non-quantified congener.

[^E0052]: Duplicate analysis: Separate analysis of the analytes of interest using a second aliquot of the same homogenised sample. In general, the requirements for duplicate analysis as provided for in Annex II, Chapter C, point 3 apply. However, for methods with the use of ¹³C-labelled internal standard for the relevant analytes, the duplicate analysis is only necessary if the result of the first determination is not compliant. The duplicate analysis is necessary to exclude the possibility of internal cross-contamination or an accidental mix-up of samples. In case the analysis is performed in the course of a contamination incident, confirmation by duplicate analysis may be omitted in case the samples selected for analysis are through traceability linked to the contamination incident and the level found is significantly above the maximum level.

[^E0053]: The concept of ‘upper-bound’ requires using the limit of quantification for the contribution of each non-quantified congener to the Toxic Equivalent (TEQ). The concept of ‘lower-bound’ requires using zero for the contribution of each non-quantified congener to the TEQ. The concept of ‘medium-bound’ requires using half of the limit of quantification calculating the contribution of each non-quantified congener to the TEQ.

[^E0054]: In general, the requirements for duplicate analysis as provided for in Annex II, Chapter C, point 3 apply. However, for confirmatory methods with the use of ¹³C-labelled internal standard for the relevant analytes, the duplicate analysis is only necessary if the result of the first determination is not compliant. The duplicate analysis is necessary to exclude the possibility of internal cross-contamination or an accidental mix-up of samples. In case the analysis is performed in the course of a contamination incident, confirmation by duplicate analysis may be omitted in case the samples selected for analysis are through traceability linked to the contamination incident and the level found is significantly above the maximum level.

[^E0055]: Identical explanation and requirements for duplicate analysis for control of action thresholds as in footnote 5 for maximum levels.

[^E0056]: Regulation (EC) No 183/2005 of the European Parliament and of the Council of 12 January 2005 laying down requirements for feed hygiene (OJ L 35, 8.2.2005, p. 1).

[^E0057]: Bioanalytical methods are not specific to those congeners included in the TEF-scheme. Other structurally related AhR-active compounds may be present in the sample extract which contribute to the overall response. Therefore, bioanalytical results cannot be an estimate but rather an indication of the TEQ level in the sample.

[^E0058]: ‘Guidance Document on Measurement Uncertainty for Laboratories performing PCDD/F and PCB Analysis using Isotope Dilution Mass Spectrometry’ (https://food.ec.europa.eu/system/files/2017-05/animal-feed-guidance_document_pcdd-f_pcb_en.pdf), ‘Guidance Document on the Estimation of LOD and LOQ for Measurements in the Field of Contaminants in Feed and Food’ (https://food.ec.europa.eu/system/files/2016-10/cs_contaminants_sampling_analysis-report_2004_en.pdf).

[^E0060]: Current requirements are based on the TEFs published in: M. Van den Berg et al, Toxicol Sci 93 (2), 223–241 (2006).

[^E0061]: Congeners often found to co-elute are for example PCB 28/31, PCB 52/69 and PCB 138/163/164. For GC-MS also possible interferences from fragments of higher chlorinated congeners shall be considered.

[^E0062]: The principles as described in the ‘Guidance Document on the Estimation of LOD and LOQ for Measurements in the Field of Contaminants in Feed and Food’ (https://data.europa.eu/doi/10.2787/8931) shall be followed when applicable.

[^E0063]: It is highly recommendable to have a lower contribution of the reagent blank level to the level of a contaminant in a sample. It is in the responsibility of the laboratory to control the variation of blank levels, in particular, if the blank levels are subtracted.

[^E0064]: See footnote 9.

[^E0066]: Regulation (EC) No 999/2001 of the European Parliament and of the Council of 22 May 2001 laying down rules for the prevention, control and eradication of certain transmissible spongiform encephalopathies (OJ L 147, 31.5.2001, p. 1).

[^E0067]: Regulation (EC) No 1069/2009 of the European Parliament and of the Council of 21 October 2009 laying down health rules as regards animal by-products and derived products not intended for human consumption and repealing Regulation (EC) No 1774/2002 (Animal by-products Regulation) (OJ L 300, 14.11.2009, p. 1).

[^E0069]: The list of these primers and probes for each animal species targeted by the assay is available on the EURL-AP website.

[^E0070]: Examples of Master Mixes that are functional are available on the EURL-AP website.