Vitamin B12 / Cyanocobalamin

Executive Summary

In the current application authorisation is sought under Articles 4 for vitamin B12 / cyanocobalamin under the category/functional group 3(a), "nutritional additives/vitamins, pro-vitamins and chemically well-defined substances having similar effect", according to the classification system of Annex I of Regulation (EC) No 1831/2003. Authorisation is sought for the use of the feed additive for all animal species and categories.
According to the Applicant, vitamin B12 / cyanocobalamin is produced by fermentation with a non-genetically modified Ensifer adhaerens CGMCC 19596 strain and further reaction with sodium cyanide to form cyanocobalamin (vitamin B12). The feed additive has a minimum vitamin B12 / cyanocobalamin purity of 96 % (w/w). Cyanocobalamin is intended to be incorporated in feedingstuffs through premixtures. No minimum or maximum contents in feedingstuffs have been proposed by the Applicant however, recommended levels range from 10 to 80 g/kg compound feed, depending on the target species.
For the determination of cyanocobalamin in the feed additive the Applicant proposed the methods stated in the European Pharmacopoeia cyanocobalamin monograph (Eur. Ph. 0547).
For the determination of cyanocobalamin in premixtures and feedingstuffs, the Applicant proposed an in-house validated microbiological assay. However, in the frame of a previous vitamin B12 evaluation, this method has triggered some NRLs concerns about its applicability for the quantification of cyanocobalamin in premixtures and feedingstuffs.
Nevertheless, the EURL is aware of a ring-trial validated AOAC method based on immunoaffinity column clean-up and high performance liquid chromatography coupled to spectrophotometry (HPLC-UV) for the determination of vitamin B12 in food commodities. The method has been further verified in different feedingstuffs, leading to a relative standard deviation for repeatability (RSDr) ranging from 9.0 to 10.8 % and a relative standard deviations for intermediate precision (RSDip) ranging from 10.1 to 10.8 %.
Based on the performance characteristics available, the EURL recommends for official control (i) the European Pharmacopoeia method (Eur. Ph. 0547) based on spectrophotometry (UV/VIS) to determine vitamin B12 / cyanocobalamin in the feed additive and (ii) the reversed phase liquid chromatography method coupled to spectrophotometric detection (HPLC-UV) based on the AOAC ring-trial validated method for the determination of vitamin B12 / cyanocobalamin in feedingstuffs.
Further testing or validation of the methods to be performed through the consortium of National Reference Laboratories as specified by Article 10 (Commission Regulation (EC) No 378/2005, as last amended by Regulation (EU) 2015/1761) is not considered necessary.