Commission Delegated Regulation (EU) 2024/1229 of 20 February 2024 supplementing Regulation (EU) 2019/4 of the European Parliament and of the Council by establishing specific maximum levels of cross-contamination of antimicrobial active substances in non-target feed and methods of analysis for these substances in feed
Data as of 2026-07-04 · Compiled from an official source version. Later amendments or repeals may not be reflected; the official text prevails. · Read the official text ↗
Subject matter and scope
This Regulation establishes specific maximum levels of cross-contamination in non-target feed for the antimicrobial active substances listed in Annex II to Regulation (EU) 2019/4, and methods of analysis for those antimicrobial active substances in feed, as provided for in Article 7(3) of Regulation (EU) 2019/4.
Specific maximum levels of cross-contamination of antimicrobial active substances in non-target feed
1. The specific maximum levels of cross-contamination in non-target feed for the antimicrobial active substances listed in Annex II to Regulation (EU) 2019/4 shall be set: (a) where the last batch manufactured, processed, stored or transported before the manufacturing, processing, storage or transport of the non-target feed is medicated feed, at 1 % of the antimicrobial active substance contained in that last batch of medicated feed, relative to a moisture content of 12 % in the non-target feed; (b) where the last batch manufactured, processed, stored or transported before the manufacturing, processing, storage or transport of the non-target feed is an intermediate product, at 1 % of the antimicrobial active substance to be contained in the medicated feed derived from that last batch of intermediate product, relative to a moisture content of 12 % in the non-target feed. 2. By way of derogation from paragraph 1, the specific maximum levels of cross-contamination in non-target feed for the antimicrobial active substances listed in Annex II to Regulation (EU) 2019/4 shall be set at the limit of quantification (‘LOQ’) laid down in the Annex to this Regulation, where the non-target feed is intended for the following animals: (a) food-producing animals other than fish where the non-target feed is manufactured, processed, stored or transported after the manufacturing, processing, storage or transport of medicated feed or intermediate products intended for aquaculture; (b) animals during the production of eggs or milk intended for human consumption; (c) food-producing animals intended for slaughter in the period for slaughter corresponding to the longest withdrawal period for the target animal species.
Methods of analysis for antimicrobial active substances in feed
The reference methods of analysis for the quantification of the level of cross-contamination in non-target feed for each antimicrobial active substance listed in Annex II to Regulation (EU) 2019/4, as referred to in Article 2(1) and (2) of this Regulation, are laid down in the Annex to this Regulation. However, alternative methods of analysis may be used provided they are validated in accordance with internationally accepted scientific protocols, are suitable to detect the same or a lower LOQ as the LOQ for the same antimicrobial active substance laid down in the Annex to this Regulation and are considered as equivalent by the competent authorities of the Member States.
Entry into force and application
This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union . It shall apply from 20 May 2025.
Maximum levels of cross-contamination in non-target feed as referred to in Article 2(2) and reference methods of analysis for the quantification of the level of cross-contamination of antimicrobial active substances in feed as referred to in Article 3
ANNEX Maximum levels of cross-contamination in non-target feed as referred to in Article 2(2) and reference methods of analysis for the quantification of the level of cross-contamination of antimicrobial active substances in feed as referred to in Article 3 Chemical class Name of substance CAS number 1 EU number 2 Multi-analyte method a b c Reference analytical method 3 4 5 Maximum levels of cross-contamination in non-target feed referred to in Article 2(2) (set at the limit of quantification (LOQ)) (μg/kg) Polymixins (polypeptide antibiotics) Colistin 1264-72-8 — (b) LSE – A – C – SPE – E – LC-MS/MS 150 (Colistin A) 300 (Colistin B) Pyrimidine inhibitor of dihydrofolate reductase Trimethoprim 738-70-5 212-006-2 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 25 Macrolides Tylvalosin 63409-12-1 — (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 Tilmicosin 108050-54-0 639-676-2 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 Tylosin 1401-69-0 215-754-8 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 Lincosamides Lincomycin 154-21-2 205-824-6 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 25 Pleuromutilins Tiamulin 55297-96-6 259-580-0 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 10 Valnemulin 101312-92-9 — (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 50 Penicillins Amoxicillin 26787-78-0 612-127-4 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 150 Penicillin V 1098-87-9 — (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 50 Aminoglycosides Apramycin 65710-07-8 265-890-7 253-460-1 (a) LSE – A – C – SPE – LC-MS/MS 50 Neomycin 1404-04-2 1404-04-2 (a) LSE – A – C – SPE – LC-MS/MS 50 Paromomycin 1263-89-4 — (a) LSE – A – C – SPE – LC-MS/MS 50 Spectinomycin 1695-77-8 — (a) LSE – A – C – SPE – LC-MS/MS 500 Amphenicols Florfenicol 73231-34-2 642-986-0 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 150 Tiamfenicol 15318-45-3 239-355-3 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 200 Coccidiostats Amprolium 137-88-2 204-458-4 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 (a) LSE – A – C – SPE – LC-MS/MS 5 Fluoroquinolones Flumequine 42835-25-6 255-962-6 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 25 Oxolinic Acid 14698-29-4 238-750-8 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 25 Sulphonamides Sulfamonomethoxine 1220-83-3 624-483-8 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 25 Sulfadimethoxine 122-11-2 204-523-7 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 25 Tetracyclines Chlortetracycline 57-62-5 200-341-7 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 Doxycycline 564-25-0 209-271-1 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 Tetracycline 60-54-8 200-481-9 (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 Oxytetracycline 79-57-2 — (c) LSE – US – A – C – F – LC-MS/MS or LC-HRMS 100 (1) Chemical Abstracts Service. (2) European Union number – not available for all substances. (3) Extraction methods: — LSE – liquid solid extraction, — US – Ultrasonication, — A – Agitation. (4) Clean-up methods: — C – Centrifugation, — SPE – Solid Phase Extraction, — E – Evaporation, re-dissolution, — F – Filtration. (5) Analytical methods: — LC-MS/MS – Liquid chromatography tandem mass spectrometry, — LC-HRMS – Liquid chromatography coupled to high resolution mass spectrometry. (a): multi-analyte method for the aminoglycosides and amprolium. (b): multi-analyte method for the polymixins colistin A and B. (c): multi-analyte method for trimethoprim, amprolium, lincomycin, the macrolides, the pleuromutilins, the penicillins, the amphenicols, the fluoroquinolones, the sulphonamides and the tetracyclines.
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