Menadione sodium bisulphite or ‘Vitamin K3 (3a710); Menadione nicotinamide bisulphite or Vitamin K3 (3a711)

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2015/2307

For the determination of menadione sodium bisulphite in the feed additive and for the determination menadione nicotinamide bisulphite in the feed additive:

- a spectrophotometric method using a visible detector at 635 nm (VDLUFA -Bd. III 13.7.1)

For the determination of menadione sodium bisulphite in premixtures and feedingstuffs and for the determination of menadione nicotinamide bisulphite in premixtures and feedingstuffs:

- Normal Phase High-Performance Liquid Chromatograph coupled to UV detector-decree 29/04/2010, Official Italian Journal No 120 25/5/2010

Executive Summary

In the current application authorisation is sought for Vitamin K3 (Menadione) under the category/functional group 3(a) ‘nutritional additives’/‘vitamins, pro-vitamins and chemically well-defined substances having similar effect’ according to Annex I of Regulation (EC) No 1831/2003. Specifically, authorisation is sought to use the active substances, Menadione Sodium Bisulphite (MSB) and Menadione Nicotinamide Bisulphite (MNB) for all animal species and categories, with a minimum content of 50 % and 44 % of Menadione, respectively. The feed additive is intended to be incorporated in feedingstuffs through premixtures or directly in water (only for MSB). However, the Applicant did not specify any maximum or minimum concentration of MSB and MNB in feedingstuffs or water.

For the determination of Vitamin K3 (Menadione) and its commercial derivatives Menadione Sodium Bisulphite (MSB) and Menadione Nicotinamide Bisulphite (MNB) in the feed additive the Applicant proposed a Method from the Association of German Agricultural Analytical Research Institutes (VDLUFA), validated for premixtures and feedingstuffs. The Applicant applied this method to the feed additive and reported a relative standard deviation of repeatability (RSDr) of 1.7 % for MSB and MNB containing 50 % and 44 % of Menadione, respectively. Moreover, a recovery rate (RRec) ranging from 101 % to 104 % was calculated by EURL, based on the experimental data provided by Applicant. Based on the performance characteristic presented, the EURL recommends for official control the validated method from VDLUFA (Bd.III 13.7.1) to determine Vitamin K3 in the feed additive.

For the determination of Vitamin K3 in premixtures and feedingstuffs the Applicant proposed the above mentioned VDLUFA method - validated for premixtures and feedingstuffs, for which an RSDr ranging from 10 % to 20 % is reported. An alternative ring-trial validated method for premixtures and feedingstuffs was published in the Italian Official Journal, based on Normal Phase High-Performance Liquid Chromatography (NP-HPLC) coupled to Ultraviolet (UV) detector, using a wavelength of 251 nm. The following performance characteristics were reported for premixtures, complementary feedingstuffs and complete feedingstuffs samples with a Vitamin K3 (Menadione) content ranging from 5.57 to 543 mg/kg: - RSDr ranging from of 2.9 to 6.8 %; - a relative standard deviation of reproducibility (RSDR) ranging from 7.0 to 11.7%; - RRec ranging from 89.2 to 111.4 %, and - a limit of detection (LOD) and quantification (LOQ) of 1.2 and 3.8 mg/kg, respectively.

Based on the performance characteristics presented, the EURL recommends for official control the ring-trial validated method, published in the Italian Official Journal, based on Normal Phase High-Performance Liquid Chromatography (NP-HPLC) coupled to UV detector to determine Vitamin K3 in premixtures and feedingstuffs, within the concentration range covered.

For the determination of the Menadione Sodium Bisulphite (MSB) in water, the Applicant applied the VDLUFA method. The following performances characteristics were reported for a concentration of 0.5 g Menadione /L: - RSDr ranging from 1.8 to 4.4 %, and - RRec ranging from 97.9 to 99.2 %.

Based on the performance characteristics presented, the EURL recommends for official control the validated method from VDLUFA (Bd.III 13.7.1) to determine Vitamin K3 in 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) is not considered necessary.