L-lysine base 3c320, L-lysine monohydrochloride 3c322ii, L-lysine sulphate 3c325i, L-lysine sulphate 3c324i, L-lysine monohydrochloride 3c322i, L-lysine sulphate 3c323

In the current applications authorisation is sought under Article 4(1) for L-lysine monohydrochloride, concentrated liquid L-lysine and l-lysine sulphate produced by Corynebacterium glutamicum KCCM80183 and L-lysine monohydrochloride produced by Corynebacterium glutamicum DSM32932, 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. Authorisation is sought for all animal species. 

According to the Applicants, L-lysine monohydrochloride contains a minimum (mass fraction) of 78 % of L-lysine as active substance, while the concentrated liquid L-lysine and the L-lysine sulphate contain a minimum of 50 and 55 % of L-lysine, respectively. 

The different forms of the feed additive are intended to be added directly into feedingstuffs or through premixtures. L-lysine monohydrochloride, concentrated liquid L-lysine and l-lysine sulphate produced by Corynebacterium glutamicum KCCM80183 can also be included in water for drinking. However the Applicants did not propose any minimum or maximum content of L-lysine in feedingstuffs. 

For the quantification of lysine in the feed additive, the Applicants submitted the European Union (EU) method dedicated for the determination of amino acids in premixtures and feedingstuffs. However, for the quantification of lysine in the feed additive the EURL previously evaluated and recommended the ring-trial validated method EN ISO 17180:2013 based on ion-exchange chromatography coupled to visible or fluorescence detection (IEC-VIS/FLD). This standard method does not distinguish between the salts of amino acids and it cannot differentiate between enantiomers. It applies for products containing more than 10 % of amino acid. The following performance characteristics are reported: a relative standard deviation for repeatability (RSD_r) ranging from 0.7 to 1.7 % and a relative standard deviation for reproducibility (RSD_R) ranging from 1.5 to 2.5 %. 

For the quantification of L-lysine in premixtures and feedingstuffs one Applicant submitted the ring-trial validated European Union method (Commission Regulation (EC) No 152/2009) based on IEC coupled with photometric detection (IEC-VIS), which was previously recommended by the EURL. This method, designed only for the analysis of amino acids in premixtures and feedingstuffs, does not distinguish between the salts and the amino acid enantiomers. The following performance characteristics were reported for the quantification of total lysine: RSD_r ranging from 2.1 to 2.8 % and RSD_R ranging from 3 to 6.7 %. 

The different forms of Lysine produced by Corynebacterium glutamicum KCCM80183 can also be included in water for drinking. However, the corresponding Applicant did not provide any experimental data to determine lysine in water. Nevertheless, as concluded in previous amino acids reports of the EURL, the IEC-VIS procedure described in the European Union method is considered fit-for-purpose for the determination of lysine in water. 

In addition, the EURL found the "L-lysine monohydrochloride monograph" of the Food Chemical Codex (FCC) for the identification of L-lysine monohydrochloride in the feed additive and the generic European Pharmacopoeia monograph (Ph. Eur. 20301) for the identification of sulphate ion in L-lysine sulphate. 

In the frame of these authorisations the EURL recommends for official control (i) the "L-lysine monohydrochloride monograph" of the Food Chemical Codex (FCC) based on infrared absorption for the identification of L-lysine monohydrochloride in the feed additive; (ii) the European Pharmacopoeia monograph (Ph. Eur. 01/2008:20301) for the identification of the sulphate ion in L-lysine sulphate; (iii) the ring-trial validated method EN ISO 17180:2013 based on ion-exchange chromatography coupled to visible or fluorescence detection (IEC-VIS/FLD) to quantify free lysine in the feed additive and premixtures (containing more than 10 % lysine); and (iv) the European Union method based on IEC-VIS for the quantification of lysine in premixtures, feedingstuffs and water. 

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) 2021/2095

For the identification of L-lysine monohydrochloride in the feed additive:

— Food Chemical Codex ‘L-lysine monohydrochloride monograph’

For the identification of sulphate in the feed additive:

—  European Pharmacopoeia Monograph 20301

For the quantification of lysine in the feed additive and premixtures containing more than 10 % lysine:

— ion exchange chromatography coupled with post-column derivatisation and optical detection (IEC-VIS/FLD) – EN ISO 17180

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

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

For the quantification of lysine in water:

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

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