L-histidine monohydrochloride monohydrate 3c352i

In the current application an authorisation is sought under Article 4(1) for L-histidine monohydrochloride monohydrate produced by fermentation with the strain Escherichia coli KCCM 80212 (H010), under the category/functional group 3(c) 'nutritional additives'/'amino acids, their salts and analogues', according to Annex I of Regulation (EC) No 1831/2003. The authorisation is sought for all animal species. 

According to the Applicant, L-histidine monohydrochloride monohydrate has a minimum purity (mass fraction) of 98 %. The feed additive is intended to be added directly into feedingstuffs or through premixtures. However, the Applicant did not propose any minimum or maximum content of L-histidine monohydrochloride monohydrate in feedingstuffs. 

For the quantification of histidine in the feed additive and premixtures the Applicant proposed and submitted the ring-trial validated method EN ISO 17180 originally dedicated for the determination of lysine, methionine and threonine in commercial amino acid products and premixtures. This standard method is based on ion-exchange chromatography coupled to visible or fluorescence detection (IEC-VIS/FLD). It does not distinguish between the salts of amino acids and it cannot differentiate between enantiomers. The method is applicable for products and premixtures containing more than 10 % in mass fraction of the amino acid. 

The extension of scope of the EN ISO 17180 method to another almost identical feed additive and premixtures containing histidine has been demonstrated by the Applicant in the frame of a recent histidine dossier. 

For the quantification of histidine in feedingstuffs the Applicant submitted the ring-trial validated European Union method (Commission Regulation (EC) No 152/2009) based on IEC coupled to photometric detection (VIS). The method is designed for the analysis of amino acids in premixtures and feedingstuffs and does not distinguish between the salts and the amino acid enantiomers. This method was further ring-trial validated by 23 laboratories, resulting in the EN ISO 13903 method. The following performance characteristics were reported for the quantification of total histidine: a relative standard deviation for repeatability (RSD_r) ranging from 2.4 to 7.0 % and a relative standard deviation for reproducibility (RSD_R) ranging from 13 to 23 %. 

Based on the performance characteristics available, the EURL recommends for official control: (i) the ring-trial validated method EN ISO 17180 based on IEC-VIS/FLD for the quantification of histidine in the feed additive and premixtures; and (ii) the ring-trial validated EU method, based on IEC-VIS for the quantification of histidine in premixtures and 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.

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2021/709

For the quantification of histidine in the feed additive:

— high performance liquid chromatography coupled with photometric detection (HPLC-UV)

— ion exchange chromatography coupled with post-column derivatisation and optical detection (IEC-VIS/FLD)

For the quantification of histidine in premixtures, feed materials and compound feed:

— ion exchange chromatography coupled with post-column derivatisation and photometric detection (IEC-VIS), Commission Regulation (EC) No 152/2009 (Annex III, F)

For the quantification of histamine in the feed additive:

— high performance liquid chromatography coupled to a spectrophotometric detection (HPLC-UV)