COMMISSION DELEGA TED REGUL ATION (EU) 2024/1229 
of 20 Febr uary 2024
supplementing Regulation (EU) 2019/4 of the European Parliament and of the Council by 
establishing specif ic maximum levels of cross-cont amination of antimicrobial activ e subst ances in 
non-target feed and methods of analysis for these subst ances in feed 
(Text with EEA relevance) 
THE EUR OPEAN COMMISSION,
Havi ng regar d to the Treaty on the Functioning of the European Union,
Havi ng regard to Regulation (EU) 2019/4 of the European Parliament and of the Council of 11 December 2018 on the 
manufact ure, 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 (1), and in particular Article 7(3) thereof,
Whereas:
(1) Regulation (EU) 2019/4 lays down specifi c provisions regar ding medicate d feed and interm ediate products. Cross- 
contamination of non-target feed with antimicrobials has been identified as a core issue of the Union in the cont ext 
of prote cting animal health, human health and the environment, and should be avoided or kept as low as possible.
(2) In accordance with Article 7(3) of Regulation (EU) 2019/4, the Commission must adopt delegat ed acts to 
supplement that Regulation by establishing, as regar ds the 24 antimicrobial active substances listed in Annex II 
thereto (‘the 24 antimicrobial active substances’), specific maximum levels of cross-contamination for the 
antimicrobial active substances in non-target feed and methods of analysis for the antimicrobial active substances in 
feed. Pursuant to Article 7(3) of that Regulation, those delega ted acts whic h establish maximum levels of cross- 
contamination must be based on a scientific risk assessment carried out by the European Food Safety Author ity 
(‘EFSA’ ).
(3) At the Commission’s request, EFSA assessed, in cooperation with the European Medicines Age ncy (‘EMA’), the 
specifi c concentrations of the 24 antimicrobial active substances resulting from cross-contamination in non-target 
feed for food-producing animals, below which there would be no effect on the emerg ence of, and/or selection for, 
resistance in antimicrobial active substances relevant for human and animal health (‘antimicrobial resistance’, ‘AMR’).
(4) EFSA was also requeste d by the Commission to assess the levels of the 24 antimicrobial active substances which 
could have a growth promotion or increased yield effect, taking into account that the use of antibiotics as feed 
additives, other than coccidiostats or histo monostats, has been phased out since 1 Januar y 2006 in accordance with 
Article 11(2) of Regulation (EC) No 1831/2003 of the European Parliament and of the Council (2). The specifi c 
maximum level of each antimicrobial active substance in non-target feed should be below the level that causes a 
growth promotion or increased yield effect.
(5) In addition, the Commission request ed the Reference Laboratory , set up pursuant to Regulation (EC) No 1831/2003 
(‘the Reference Laboratory’), to recommend methods of analysis for the 24 antimicrobial active substances in feed.
Offi cial Jour nal 
of the European Union EN 
L series 
2024/1229 30.4.2024
ELI: http://data.europa.eu/eli/reg_del/2024/1229/oj 1/7(1) OJ L 4, 7.1.2019, p. 1, ELI: http://data.europa.eu/eli/reg/2019/4/oj.
(2) Regulation (EC) No 1831/2003 of the European Parliament and of the Council of 22 Sept ember 2003 on additives for use in animal 
nutr ition (OJ L 268, 18.10.2003, p. 29, ELI: http://data.europa.eu/eli/reg/2003/1831/2021-03-27).(6) In its 13 Opinions of 15 Sept ember 2021 on maximum levels of cross-contamination for the 24 antimicrobial active 
substances in non-target feed (3)(‘Opinions of 15 Sept ember 2021 ’), EFSA could only establish specifi c 
concentrations concer ning AMR for six of the 24 antimicrobial active substances and not for all relevant animal 
species, due to a lack of data. In addition, EFSA only identif ied levels causing effects on growth promotion or 
increased yield for 14 of the 24 antimicrobial active substances and not for all relevant animal species, again due to 
an absence of relevant data.
(7) In Apr il 2022 and Febr uary 2023, the Reference Laborat ory issued two repor ts on the methods of analysis and 
minimum achi evable limits of quantifi cation (‘LOQ’) in feed for the 24 antimicrobial active substances (4)(‘Repor ts 
of Apr il 2022 and Febr uary 2023’).
(8) The specifi c concentrations concer ning AMR established by EFSA for six antimicrobial active substances, in the 
Opinions of 15 September 2021 , are signif icantly lower than the minimum LOQs established by the Reference 
Laboratory in the Repor ts of Apr il 2022 and Febr uary 2023. This means, in practice, that the specifi c 
concentrations are not measurable and would, theref ore, not be enforceable by the Member States in accordance 
with Article 17(2) of Regulation (EC) No 178/2002 of the European Parliament and of the Council (5).
(9) The lowe st levels of the 14 antimicrobial active substances, for whic h EFSA could indicate in its Opinions of 
15 September 2021 as causing a growth promotion or increased yield effect, are signif icantly higher than the LOQ 
for the same substance and are theref ore measurable and enforceable by the Member States in accordance with 
Article 17(2) of Regulation (EC) No 178/2002. To avoid a growth promotion or increased yield effect, the 
maximum levels of cross-contamination for the antimicrobial active substances in non-target feed should be below 
the lowest levels causing a growth promotion or increased yield effect.
(10) High economic inve stment and increased logistical costs to compl y with the maximum levels of cross- 
contamination in non-target feed if such levels are very low is likely to result in a reduction of the production of 
medicate d feed. In addition, the EMA Advice of 28 August 2020 on implementing measures under Article 106(6) 
of Regulation (EU) 2019/6 of the European Parliament and of the Council (6)on veterinar y medicinal products – 
scientific problem analysis and recommendations to ensure a safe and efficient administration of oral veteri nary 
medicinal products via routes other than medicated feed(7), concludes that it may also result in an increased 
recourse to methods of oral administration of antimicrobial active substances other than medicated feed, such as 
the administration on the surface of solid feed, that may increase the risk of AMR and the inability to treat certain 
bacterial infections in certain species due to the absence of other appropr iate route s of administration, for example, 
in aquaculture. The maximum levels of cross-contamination should, theref ore, not be detr imental to the production 
of medicate d feed, in particular , by small and medium-sized feed manufactur ing plants, excl uding them in practice 
from the production of medicated feed, which would result in possible issues for public health, and animal health 
and welf are. It is, theref ore, appropr iate to establish a maximum level of cross-contamination that is strict but also 
feasible to achi eve by applying good practices to minimise cross-contamination. In addition to the Opinions of 
15 September 2021 , the exper ience gained in the Member State s in applying national law indicate s that a cross- 
contamination level in the non-target feed of 1 % of the active substance in the medicated feed, represents a good 
balance between feasibility and AMR control. Interm ediat e products contain higher concentrations of active 
substances than medicate d feed. Theref ore, where non-target feed is manufactured, processed, stored or transpor ted 
after the manufactur ing, processing storage or transpor t of interm ediate products, a cross-contamination level of 
1 % of the substance to be contained in the derived medicat ed feed, should apply .EN OJ L, 30.4.2024 
2/7 ELI: http://data.europa.eu/eli/reg_del/2024/1229/oj(3) EFSA Journal 2021;19(10):6852 to 6865.
(4) Vincent, U., Oliveira Gonçalves, C., Ferrari, L., Bouten, K., Chedin, M., Stroka, J., Pinotti, L. and Von Holst, C., Determ ination of 24 
antibiotics at trace levels in animal feed by High Perfor mance Liquid Chromatograph y – Tandem Mass Spectrometr y (LC- MS/MS), 
Publications Offi ce of the European Union, Luxembourg, 2024, EUR 31818 EN, doi:10.2760/12878, JRC136836.
(5) Regulation (EC) No 178/2002 of the European Parliament and of the Council of 28 Januar y 2002 laying down the general principles 
and requirements of food law, establishing the European Food Safet y Authority and laying down procedures in matt ers of food safety 
(OJ L 31, 1.2.2002, p. 1, ELI: http://data.europa.eu/eli/reg/2002/178/oj).
(6) Regulation (EU) 2019/6 of the European Parliament and of the Council of 11 December 2018 on veterin ary medicinal products and 
repealing Directive 2001/82/EC (OJ L 4, 7.1.2019, p. 43, ELI: http://data.europa.eu/eli/reg/2019/6/oj).
(7) EMA/CVMP/508559/2019.(11) The maximum levels of cross-contamination for some antimicrobial active substances in non-targe t feed should be 
reviewed if new scientifi c evidence becomes available, allowing to further control antimicrobial resistance in the 
non-target feed with enforceable maximum levels whic h are achi evable by applying good practices to minimise 
cross-contamination.
(12) Medicated feed or interm ediate products intended for fish often contains substantially higher doses of antimicrobial 
active substances than medicat ed feed or interm ediate products intende d for food-producing animals other than fish. 
In addition, no levels of antimicrobial active substances creating a growth promotion or increased yield effect in fish, 
have been identified in the Opinions of 15 Sept ember 2021 . Stricter specifi c maximum levels of cross- 
contamination in non-target feed intended for food-producing animals other than fish theref ore are needed where 
the cross-contamination originates from medicated feed or interm ediate products intended for fish, in order to 
avoid a growth promotion or increased yield effect in food-producing animals other than fish. Since these stricter 
specifi c maximum levels of cross-contamination in non-targe t feed intended for food-producing animals other than 
fish should be measurable and enforceable by the Member State s, they should be set at the LOQ.
(13) It should be ensured that food derived from animals fed with the non-target feed compl ies with the maximum 
residue limits laid down in Table 1 set out in the Annex to Commission Regulation (EU) No 37/2010 (8). Stricter 
specifi c maximum levels of cross-contamination for antimicrobial active substances in non-target feed should, 
theref ore, be laid down in this Regulation, in particular for milk - or egg-producing animals and for animals close to 
the date of slaughter . Since these stricter specifi c maximum levels of cross-contamination in non-target feed should 
be measurable and enforceable by the Member States, they should be set at the LOQ.
(14) The methods of analysis recommended by the Reference Laborat ory in the Repor ts of Apr il 2022 and Febr uary 
2023 should be used as reference methods for the analysis of the 24 antimicrobial active substances in feed. 
Alte rnative methods of analysis should only be allowe d when validat ed and considered as equivalent by the 
compet ent author ities of the Member States.
(15) It is appropr iate to provi de official laboratories carrying out the methods of analysis for antimicrobial active 
substances in feed with suffi cient time to adapt to the LOQs and prove their compe tence for carrying out such 
methods of analysis by generally accept ed means, such as by accreditation, sound in-house validation or proficiency 
test data targeting a timely accreditation. Theref ore, this Regulation should apply 12 months after the date of its 
entr y into force,
HAS ADOPTED THIS REGUL ATION:
Article 1
Subject matter and scope
This Regulation establishes specific maximum levels of cross-contamination in non-target feed for the antimicrobial active 
substances liste d in Annex II to Regulation (EU) 2019/4, and methods of analysis for those antimicrobial active substances 
in feed, as provid ed for in Article 7(3) of Regulation (EU) 2019/4.
Article 2
Specif ic maximum levels of cross-cont amination of antimicrobial activ e subs tances in non-t arget feed
1. The specific maximum levels of cross-contamination in non-target feed for the antimicrobial active substances liste d 
in Annex II to Regulation (EU) 2019/4 shall be set:
(a) where the last batch manuf actured, processed, stored or transpor ted before the manuf actur ing, processing, storage 
or transpor t of the non-target feed is medicate d feed, at 1 % of the antimicrobial active substance contained in that 
last batch of medicat ed feed, relative to a moisture content of 12 % in the non-target feed;OJ L, 30.4.2024 EN 
ELI: http://data.europa.eu/eli/reg_del/2024/1229/oj 3/7(8) Commission Regulation (EU) No 37/2010 of 22 December 2009 on phar macologically active substances and their classification 
regar ding maximum residue limits in foodstuffs of animal origin (OJ L 15, 20.1.2010, p. 1, ELI: http://data.europa.eu/eli/reg/2010/37 
(1)/2023-06-11).(b) where the last batch manuf actured, processed, stored or transpor ted before the manuf actur ing, processing, storage 
or transpor t of the non-target feed is an interm ediate product, at 1 % of the antimicrobial active substance to be 
contained in the medicate d 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 deroga tion from paragraph 1, the specifi c 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 quantif ication 
(‘LOQ’) laid down in the Annex to this Regulation, where the non-target feed is intended for the followi ng animals:
(a) food-producing animals other than fish where the non-target feed is manufact ured, processed, stored or transpor ted 
after the manufa ctur ing, processing, stora ge or transpor t of medicated feed or interm ediat e products intended for 
aquaculture;
(b) animals during the production of eggs or milk intended for human consump tion;
(c) food-producing animals intended for slaughter in the period for slaughter corresponding to the long est withdrawal 
period for the target animal species.
Article 3
Methods of analysis for antimicrobial activ e subst ances in feed
The reference methods of analysis for the quantif ication 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 refer red to in Article 2(1) and (2) of this 
Regulation, are laid down in the Annex to this Regulation.
However , altern ative methods of analysis may be used provided they are validate d in accordance with internati onally 
accept ed scientific prot ocols, are suitable to detec t the same or a lowe r 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 compet ent author ities of the 
Member States.
Article 4
Entr y into force and application
This Regulation shall enter into force on the twentieth day followi ng that of its publication in the Official Journal of the 
European Union.
It shall apply from 20 May 2025 .
This Regulation shall be binding in its entirety and directly applicable in all Member States.
Done at Brussels, 20 Febr uary 2024 .
For the Commission
The President
Ursula VON DER LEYENEN OJ L, 30.4.2024 
4/7 ELI: http://data.europa.eu/eli/reg_del/2024/1229/ojANNEX 
Maximum levels of cross-cont amination in non-t arget feed as refer red to in Article 2(2) and reference methods of analysis for the quantif ication of the level of 
cross-contamination of antimicrobial activ e subst ances in feed as referred to in Article 3 
Chemical class Name of substance CAS number1EU number2Multi-analyte 
methoda b cReference analytical method3 4 5Maximum levels of cross- 
contamination in non-target feed 
referred to in Article 2(2) (set at the 
limit of quantificat ion (LOQ))
(μg/kg)
Polymixins (polypept ide 
antibiotics)Colistin 1264-72-8 — (b) LSE – A – C – SPE – E – LC-MS/MS 150 (Colistin A)
300 (Colistin B)
Pyrimidine inhibitor of 
dihy drof olate reductaseTrimethopr im 738-70-5 212-006-2 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS25
MacrolidesTylvalosin 63409-12-1 — (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
Tilmicosin 108050-54-0 639-676-2 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
Tylosin 1401-69-0 215-754-8 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
LincosamidesLincomy cin 154-21-2 205-824-6 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS25
PleuromutilinsTiamulin 55297-96-6 259-580-0 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS10
Valnemulin 101312-92-9 — (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS50
PenicillinsAmo xicillin 26787-78-0 612-127-4 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS150
Penicillin V 1098-87-9 — (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS50OJ L, 30.4.2024 EN 
ELI: http://data.europa.eu/eli/reg_del/2024/1229/oj 5/7Chemical class Name of substance CAS number1EU number2Multi-analyte 
methoda b cReference analytical method3 4 5Maximum levels of cross- 
contamination in non-target feed 
referred to in Article 2(2) (set at the 
limit of quantificat ion (LOQ))
(μg/kg)
Aminog lycosidesApramy cin 65710-07-8 265-890-7
253-460-1(a) LSE – A – C – SPE – LC-MS/MS 50
Neomy cin 1404-04-2 1404-04-2 (a) LSE – A – C – SPE – LC-MS/MS 50
Paromom ycin 1263-89-4 — (a) LSE – A – C – SPE – LC-MS/MS 50
Spectinom ycin 1695-77-8 — (a) LSE – A – C – SPE – LC-MS/MS 500
AmphenicolsFlor fenicol 73231-34-2 642-986-0 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS150
Tiamfenicol 15318-45-3 239-355-3 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS200
CoccidiostatsAmprolium 137-88-2 204-458-4 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
(a) LSE – A – C – SPE – LC-MS/MS 5
FluoroquinolonesFlumequine 42835-25-6 255-962-6 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS25
Oxolinic Acid 14698-29-4 238-750-8 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS25
SulphonamidesSulfamonometho xine 1220-83-3 624-483-8 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS25
Sulfadimetho xine 122-11-2 204-523-7 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS25EN OJ L, 30.4.2024 
6/7 ELI: http://data.europa.eu/eli/reg_del/2024/1229/ojChemical class Name of substance CAS number1EU number2Multi-analyte 
methoda b cReference analytical method3 4 5Maximum levels of cross- 
contamination in non-target feed 
referred to in Article 2(2) (set at the 
limit of quantificat ion (LOQ))
(μg/kg)
Tetracy clinesChlor tetracy cline 57-62-5 200-341-7 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
Doxycycline 564-25-0 209-271-1 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
Tetracy cline 60-54-8 200-481-9 (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
Oxyt etracy cline 79-57-2 — (c) LSE – US – A – C – F – LC-MS/MS or 
LC-HRMS100
(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 – Centr ifugation,
— SPE – Solid Phase Extraction,
— E – Evaporation, re-dissolution,
— F – Filtration.
(5) Analytical methods:
— LC-MS/MS – Liquid chromatograph y tandem mass spectrometr y,
— LC-HRMS – Liquid chromatograph y coupled to high resolution mass spectrometr y.
(a): multi-analyt e method for the aminog lycosides and ampr olium.
(b): multi-analyt e method for the polymixins colistin A and B.
(c): multi-analyt e method for trimethopr im, amproliu m, lincom ycin, the macrolides, the pleuromutilins, the penicillins, the amph enicols, the fluoroquinolones, the sulphonamides and 
the tetracyclines.OJ L, 30.4.2024 EN 
ELI: http://data.europa.eu/eli/reg_del/2024/1229/oj 7/7