COMMISSION IMPLEMENTING REGUL ATION (EU) 2023/651 
of 20 March 2023
concer ning the author isation of ribof lavin (vitamin B2) produced by Bacillus subtilis KCCM 10445 
and a preparation of ribof lavin produced by Bacillus subtilis KCCM 10445 as feed additiv es for all 
animal species 
(Text with EEA relevance) 
THE EUR OPEAN COMMISSION,
Havi ng regar d to the Treaty on the Functioning of the European Union,
Havi ng rega rd to 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 (1), and in particular Article 9(2) thereof,
Whereas:
(1) Regulation (EC) No 1831/2003 provid es for the author isation of additives for use in animal nutr ition and for the 
grounds and procedures for granting such an author isation.
(2) In accordance with Article 7 of Regulation (EC) No 1831/2003, an application was submitted for the author isation 
of ribof lavin 98 % (vitamin B2) produced by Bacillus subtilis KCCM 10445 and of a preparation of ribof lavin 80 % 
produced by Bacillus subtilis KCCM 10445. That application was accompanied by the particulars and documents 
required under Article 7(3) of Regulation (EC) No 1831/2003.
(3) The application concer ns the author isation of ribof lavin produced by Bacillus subtilis KCCM 10445 and of a 
preparation of ribof lavin produced by Bacillus subtilis KCCM 10445 as feed additives for all animal species, to be 
classifie d in the categor y ‘nutr itional additives ’ and in the functional group ‘vitamins, pro-vitamins and chemically 
well-def ined substances having similar effect’.
(4) The European Food Safe ty Auth ority (‘the Authority’) concluded in its opinions of 5 May 2021 (2)and of 
27 Sept ember 2022 (3)that, under the proposed conditions of use, ribof lavin (98 %) produced by Bacillus subtilis 
KCCM 10445 and the preparation of ribof lavin (80 %) produced by Bacillus subtilis KCCM 10445 do not have 
adverse effects on animal health, consumer safet y or the environment. It further concluded that the preparation of 
ribof lavin is not hazardous by inhalation. In the absence of data, no conclusions on the possible risk by inhalation 
of ribof lavin produced by Bacillus subtilis KCCM 10445 can be reached. Neither ribof lavin nor the preparation of 
ribof lavin are irritant to skin or eyes. In addition, in the absence of data, no conclusions on the skin sensitisation 
poten tial of ribof lavin produced by Bacillus subtilis KCCM 10445 and of the preparation of ribof lavin produced by 
Bacillus subtilis KCCM 10445 can be reached.
(5) The Author ity concluded that ribof lavin produced by Bacillus subtilis KCCM 10445 and the preparation of ribof lavin 
produced by Bacillus subtilis KCCM 10445 are an effective source in covering the nutr itional needs of animals when 
administe red via feed and/or water for drinking. The Author ity does not consider that there is a need for specifi c 
requirements of post-market monitor ing. It also verified the repor t on the methods of analysis of the feed additives 
in feed submitte d by the Reference Laboratory set up by Regulation (EC) No 1831/2003.
(6) An author isation of ribof lavin with a purity of minimum 80 % produced by Bacillus subtilis KCCM 10445 was 
previously denied and existing stoc ks of that additive and feed containing it were to be withdrawn from the mark et 
as laid down by Commission Implementing Regulation (EU) 2018/1254 (4), due to the presence in the additive of 
viable cells and recombinant DNA from the genetically modified production strain Bacillus subtilis KCCM 10445, 
(1)OJ L 268, 18.10.2003, p. 29.
(2)EFSA Journal 2021;19(6):6629.
(3)EFSA Journal 2022;20(10):7607.
(4)Commission Implementing Regulation (EU) 2018/1254 of 19 September 2018 concer ning the denial of author isation of ribof lavin 
(80 %) produced by Bacillus subtilis KCCM-10445 as a feed additive belonging to the functional group of vitamins, pro-vitamins and 
chemically well-def ined substances havin g similar effec t (OJ L 237, 20.9.2018, p. 5).EN Offi cial Jour nal of the European Union 21.3.2023 L 81/19  carrying antimicrobial resistance genes and posing a risk for the target species, consumers, users and the 
environment. The ribof lavin for whic h author isation was denied as well as the present ribof lavin (98 %) produced 
by Bacillus subtilis KCCM 10445 and the preparation of ribof lavin (80 %) produced by Bacillus subtilis KCCM 10445 
are produced by the same production strain (Bacillus subtilis KCCM 10445). However , the purity of ribof lavin has 
been improved, by increasing from 80 % in the author isation that was denied to 98 % in this author isation. In its 
abov e-mentioned opinions, the Author ity stated that viable cells and recombinant DNA from Bacillus subtilis KCCM 
10445 were not detected in the ribof lavin and in the preparation of ribof lavin subject to this application and that 
theref ore, those additives do not pose any safet y concer n associated with the genetic modif ication of the production 
strain.
(7) The assessment of ribof lavin produced by Bacillus subtilis KCCM 10445 and of the preparation of ribof lavin 
produced by Bacillus subtilis KCCM 10445 shows that the conditions for author isation, as provid ed for in Article 5 
of Regulation (EC) No 1831/2003, are satisfied. According ly, the use of the substance and of the preparation should 
be author ised. In addition, the Commission considers that appropr iate prote ctive measures should be take n to 
prevent adverse effects on human health, in particular as regards the users of those additives.
(8) The measures provid ed for in this Regulation are in accordance with the opinion of the Standing Committ ee on 
Plants, Animals, Food and Feed,
HAS ADOPTED THIS REGUL ATION:
Article 1
The substance and the preparation specified in the Annex, belonging to the additive categor y ‘nutr itional additives ’ and to 
the functional group ‘vitamins, pro-vitamins and chemically well-defined substances having similar effect’, are author ised 
as additives in animal nutr ition, subject to the conditions laid down in that Annex.
Article 2
This Regulation shall enter into force on the twentieth day followi ng that of its publication in the Official Journal of the 
European Union.
This Regulation shall be binding in its entirety and directly applicable in all Member States.
Done at Brussels, 20 Marc h 2023 .
For the Commission
The President
Ursula VON DER LEYENEN Offi cial Jour nal of the European Union L 81/20 21.3.2023  ANNEX 
Identification 
number of 
the additiveAdditiveCompo sition, chemical formula, descr iption, analytical 
methodSpecies or 
categ ory of 
animalMaximum 
ageMinimum 
conten tMaximum 
conten t
Other provisi onsEnd of period of 
author isationmg of 
active substance/kg of 
complet e feedingstuff 
with a moisture 
content of 12 %
Categor y of nutr itional additiv es. Functional group: vita mins, pro-vita mins and chemically well-def ined subst ances having similar effect
3a825 iii ‘Ribof lavin’ or 
‘Vitamin B2’Additive composition
Ribof lavin with a maximum of 1,5 % of water
Solid formAll animal 
species— — — 1.The additive may be used via water for 
drinking.
2.In the directions for use of the additive 
and premixtures, the storage condi ­
tions, the stability to heat treatment 
and the stability in water for drinking 
shall be indicate d.
3.For users of the additive and premix ­
tures, feed business operat ors shall es­
tablish operational procedures and 
organisational measures to address 
poten tial risks resulting from their 
use. Where those risks cannot be 
eliminated or reduced to a minimum 
by such procedures and measures, the 
additive and premixtures shall be used 
with appropr iate personal prot ective 
equipment, including skin and 
breathing prot ection.11.4.2033
Char acter isation of active substance
Chemical formula: C17H20N4O6
CAS number: 83-88-5
Purity: minimum 98 %
Produced by fermentation with Bacillus subtilis KCCM 
10445
Analytical method (1)
For the determination of ribof lavin in the feed 
additive: — European Phar macopoeia Ph. 
Eur 01/2008:0292
For the determination of ribof lavin in premixtures: 
— High Performance Liquid Chromatograph y with 
UV detec tion (HPL C-UV) — VDL UFA Bd. III, 13.9.1
For the determination of ribof lavin (as total vitamin 
B2) in compound feed and water: — High 
Performan ce Liquid Chromatograph y with 
Fluorescence detec tion (HPL CFLD) — EN 14152
(1)Details of the analytical methods are avail able at the follo wing address of the Reference Laboratory : https://ec.europa.eu/jrc/en/eurl/feed-additives/evaluation-repor tsEN Offi cial Jour nal of the European Union 21.3.2023 L 81/21  Identification 
number of 
the additiveAdditiveCompo sition, chemical formula, descr iption, analytical 
methodSpecies or 
categ ory of 
animalMaximum 
ageMinimum 
conten tMaximum 
conten t
Other provisi onsEnd of period of 
author isationmg of 
active substance/kg of 
complet e feedingstuff 
with a moisture 
content of 12 %
Categor y of nutr itional additiv es. Functional group: vita mins, pro-vita mins and chemically well-def ined subst ances having similar effect
3a825 IV ‘Ribof lavin’ or 
‘Vitamin B2’Additive composition
Preparation containing a minimum of 80 % of 
ribof lavin and a maximum of 3 % of water
Solid formAll animal 
species— — — 1.The additive may be used via water for 
drinking.
2.In the directions for use of the additive 
and premixtures, the storage condi ­
tions, the stability to heat treatment 
and the stability in water for drinking 
shall be indicate d.
3.For users of the additive and premix ­
tures, feed business operat ors shall es­
tablish operational procedures and 
organizational measures to address 
poten tial risks resulting from their 
use. Where those risks cannot be 
eliminated or reduced to a minimum 
by such procedures and measures, the 
additive and premixtures shall be used 
with appropr iate personal prot ective 
equipment, including skin protection.11.4.2033
Char acter isation of active substance
Chemical formula: C17H20N4O6
CAS number: 83-88-5
Purity: minimum 98 %
Produced by fermentation with Bacillus subtilis KCCM 
10445
Analytical method (1)
For the determination of ribof lavin in the feed 
additive: European Phar macopoeia Ph. Eur. 
monograph:0292
For the determination of ribof lavin in premixtures: 
— High Performance Liquid Chromatograph y with 
UV detec tion (HPL C-UV) — VDL UFA Bd. III, 13.9.1
For the determination of ribof lavin (as total vitamin 
B2) in compound feed and water: — High 
Performan ce Liquid Chromatograph y with 
Fluorescence detec tion (HPL CFLD) — EN 14152.
(1)Details of the analytical methods are avail able at the follo wing address of the Reference Laboratory : https://ec.europa.eu/jrc/en/eurl/feed-additives/evaluation-repor tsEN Offi cial Jour nal of the European Union L 81/22 21.3.2023