Commission Implementing Regulation (EU) 2025/2491 of 10 December 2025 concerning the authorisation of a preparation of 25-hydroxycholecalciferol produced with Saccharomyces cerevisiae CBS 146008 as a feed additive for all animal species except poultry, pigs and ruminants

• CELEX: Implementing Regulation (EU) 2025/2491
• Articles: 3

Article 1 Authorisation

The preparation specified in the Annex, belonging to the additive category ‘nutritional additives’ and to the functional group ‘vitamins, pro-vitamins and chemically well-defined substances having similar effect’, is authorised as an additive in animal nutrition, subject to the conditions laid down in that Annex.

Article 2 Entry into force

This Regulation shall enter into force on the twentieth day following that of its publication in the Official Journal of the European Union .

Schedules & Appendices

ANNEX

ANNEX

Identification number of the feed additive

Name of the Additive

Composition, chemical formula, description, analytical method.

Species or category of animal

Maximum age

Minimum content

Maximum content

Other provisions

End of period of authorisation

mg active substance/kg of complete feed with a moisture content of 12 %

Category: Nutritional additives. Functional group: Vitamins, provitamins and chemically well-defined substances having similar effect

Sub classification: vitamin D

3a670a

25-Hydroxycholecalciferol

Additive composition

Preparation with a maximum content of 1,25 % of 25-hydroxycholecalciferol

Solid form

Characterisation of the active substance

25-Hydroxycholecalciferol. Its precursor compound,5,7,24-cholestatrienol, is produced with Saccharomyces cerevisiae CBS 146008. After extraction, the precursor is converted chemically to 25-hydroxy-pro-vitamin D 3 , which is further transformed photochemically to 25-hydroxycholecalciferol.

Chemical formula: C 27 H 44 O 2 •H 2 O

CAS number: 63283-36-3

Purity criteria:

—

25-hydroxycholecalciferol > 94 %;

—

other sterol derivatives ≤ 1 % each;

—

erythrosine < 5 mg/kg.

Analytical method

( 1 )

For the determination of 25-hydroxycholecalciferol in the feed additive: Ultra Performance Liquid Chromatography coupled to spectrophotometric detection (UPLC-UV).

For the determination of 25-hydroxycholecalciferol in premixtures: High Performance Liquid Chromatography coupled to spectrophotometric detection (HPLC-UV).

For the determination of 25-hydroxycholecalciferol in compound feed and in low concentrated premixtures: High Performance Liquid Chromatography coupled to tandem mass spectrometry (HPLC-MS/MS).

Salmonids

0,800

1.

The additive shall be incorporated in feedingstuffs via the use of a premixture.

2.

In the directions for use of the additive and premixtures, the storage conditions and the stability to heat treatment shall be indicated.

3.

Maximum content of the combination of 25-hydroxycholecalciferol with cholecalciferol (vitamin D 3 ) per kg of complete feedingstuff:

—

≤ 1,500 mg  ( 2 ) (equivalent to 60 000 IU of cholecalciferol) for salmonids,

—

≤ 0,075 mg  ( 2 ) (equivalent to 3 000 IU of cholecalciferol) for fish species other than salmonids and for ornamental fish,

—

≤ 0,050 mg  ( 2 ) (equivalent to 2 000 IU of cholecalciferol) for other categories of animals except poultry, pigs and ruminants.

4.

For users of the additive and premixtures, feed business operators shall establish operational procedures and organisational measures to address the potential risks resulting from their use. Where those risks cannot be eliminated by such procedures and measures, the additive and premixtures shall be used with personal breathing and skin protective equipment.

31 December 2035

Finfish species other than salmonids

0,075

Ornamental fish

0,075

Other species and categories of animals except pigs, poultry and ruminants

0,050

( 1 )   Details of the analytical methods are available at the following address of the Reference Laboratory: https://joint-research-centre.ec.europa.eu/eurl-fa-eurl-feed-additives/eurl-fa-authorisation/eurl-fa-evaluation-reports_en .

( 2 )   40 IU cholecalciferol (vitamin D3) = 0,001 mg cholecalciferol (vitamin D 3 ).