- Publication date
- 20 May 2022 (Last updated on: 8 July 2022)
- Joint Research Centre
In the current application an authorisation is sought under Article 4 for lemon extract (Citrus limon Burm) under the category / functional group (4 d) "zootechnical additives"/"other zootechnical additives", according to the classification system of Annex I of Regulation (EC) No 1831/2003. Specifically, the authorisation is sought for the feed additive to be used for weaned piglets and all growing poultry species.
The Applicant proposed as phytochemical markers for the characterisation of the feed additive (i) total polyphenols and (ii) eriocitrin. The product contains respectively a minimum content of 1 % of total polyphenols and 4000 mg/kg product of eriocitrin. The feed additive (Citrozest®) is a liquid extract from the peel and the seeds of Citrus limon (L.) Burm. fruit. The product is to be used through premixtures or directly into feedingstuffs at proposed inclusion levels ranging from a minimum of 120 to a maximum 1000 mg/kg feedingstuffs.
For the determination of the total polyphenols in the feed additive the Applicant proposed a colourimetric method described in the European Pharmacopoeia monograph 2.8.14. Upon request of the EURL, in the frame of the similar “FAD-2010-0130 – Lemon extract” dossier, the Applicant provided experimental data for the analysis of the total polyphenols content in two different batches of lemon extract. Based on the acceptable performance characteristics presented the EURL recommends for official control the colourimetric method described in the European Pharmacopoeia 2.8.14 monograph for the determination of the total polyphenols content in the feed additive.
For the determination of the eriocitrin content in the feed additive the Applicant presented an extension of scope of the ring-trial validated method from the “International Fruit and Vegetable Association” (IFU 580) dedicated to the determination of hesperidin and naringin in juices. In the frame of the similar “FAD-2010-0130 – Lemon extract” dossier, the Applicant provided experimental data for the analysis of the eriocitrin content in two different batches of lemon extract. According to the results provided, the EURL calculated a relative standard deviation for repeatability (RSDr) of 0.1 % and 1.8 %, and a relative standard deviation for intermediate precision (RSDip) of 8.1 % and 8.5 %. Based on the experimental evidences provided, the EURL considers the above mentioned HPLC-UV method fit-for-purpose for the determination of the eriocitrin content in the feed additive.
For the determination of the eriocitrin content in premixtures the Applicant submitted a single-laboratory validated and further verified method based on HPLC-UV. For the determination of the eriocitrin content in feedingstuffs the Applicant submitted a single-laboratory validated and further verified method based on ultra-high performance liquid chromatography electrospray ionization tandem mass spectrometry (UPLC-ESI-MS/MS). Based on the experimental evidences provided, the EURL considers the proposed HPLC-UV and UPLC-ESI-MS/MS methods fit-for-purpose for the potential official control of eriocitrin content respectively in premixtures and feedingstuffs.
Furthermore, the Applicant did not provide experimental data or analytical methods for the determination of lemon extract in premixtures and feedingstuffs, as the unambiguous determination of the feed additive added to the matrices is not achievable experimentally. Therefore, the EURL cannot evaluate nor recommend any method for official control for the determination of lemon extract 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.