L-leucine produced by fermentation with Corynebacterium glutamicum KCCM 80421

In the current application an authorisation is sought under Article 4 for L-leucine produced by fermentation with Corynebacterium glutamicum KCCM 80421, under the category/functional groups 3(c) 'nutritional additives'/'amino acids, their salts and analogues' and 2(b) 'sensory additives'/'flavouring compounds' according to Annex I of Regulation (EC) No 1831/2003. Specifically, the authorisation is sought for all animal species. According to the Applicant, L-leucine has a minimum purity (mass fraction) of 90 %. The feed additive is intended to be mixed either into premixtures, incorporated through complementary feed or added directly to compound feed or water. The Applicant did not propose any minimum or maximum content of L-leucine in compound feed or water when used as nutritional additive, however a maximum recommended inclusion level of 25 mg/kg compound feed was proposed when used as sensory additive. For the determination of leucine in the feed additive, premixtures, compound feed and water the Applicant proposed the ring-trial validated European Union (EU) method (Commission Regulation (EC) No 152/2009). The method is based on ion-exchange chromatography coupled to optical detection (IEC-VIS) and is dedicated for the analysis of free and total amino acids in premixtures and compound feed. The following combined performance characteristics were reported for the determination of total leucine in animal feedingstuffs: a relative standard deviation for repeatability (RSD_r) ranging from 1.9 to 2.7 % and a relative standard deviation for reproducibility (RSD_R) ranging from 6.3 to 7.6 %. Furthermore, the Applicant, in the frame of the batch-to-batch variation studies, provided experimental data demonstrating the applicability of the European Union (EU) method for the analysis of leucine in the feed additive. Moreover, the EURL is aware on the suitability of the abovementioned method for the determination of leucine in water. Based on the performance characteristics available, the EURL recommends for official control the ring-trial validated European Union (EU) method, based on IEC-VIS to determine leucine in the feed additive, premixtures, compound feed and water. However, when the feed additive is intended to be used as 'sensory additives'/'flavouring compounds', the maximum recommended inclusion level of leucine in compound feed is lower than the typical limit of quantification reported for the EU method, thus the EURL is unable to recommend the EU method for the official control. Moreover, for the determination of leucine in the feed additive and premixtures, the EURL is also aware on the suitability of the ring-trial validated method EN ISO 17180 (listed in Commission Regulation (EC) No 152/2009), dedicated for the determination of free lysine, methionine and threonine in commercial amino acid products and premixtures. The method is based on IEC with optical (visible - VIS or fluorescence - FLD) detection. It does not distinguish between the salts of amino acids and cannot differentiate between enantiomers. It is applicable for products containing more than 10 % of the amino acid. Based on the performance characteristics available, the EURL recommends for official control the EN ISO 17180 method for the determination of free leucine in the feed additive and premixtures (containing more than 10 % of the amino acid). 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.