Nicotinic acid; Niacinamide

In the current application authorisation is sought under article 4(1)^1,2 (new use in water) and 10(2)^1-5 for Niacin (Nicotinic acid) and Niacinamide (Nicotinamide) under the category/functional group 3(a) "nutritional additives"/"vitamins, pro-vitamins and chemically well-defined substances having similar effect", according to the classification system of Annex I of Regulation (EC) No 1831/2003. Authorisation is sought for the use of the feed additive for all animal species and categories.

According to the Applicants the products are white granular crystals or crystalline powder, containing a minimum of 99³ % Niacin and a minimum of 98.5³ % Niacinamide. The feed additive is intended to be processed in premixtures or added directly into feedingstuffs or in water. However, the Applicants did not specify any maximum or minimum concentration of Niacin and Niacinamide in feedingstuffs or water, as set in the previous regulations.

For the determination of Niacin and Niacinamide in the feed additive, the Applicants propose various methods including the internationally recognised European Pharmacopoeia methods (Ph. Eur. 6th Edition, monograph 0459 for the determination of Niacin and monograph 0047 for the determination of Niacinamide). Even though no performance characteristics are provided, the EURL recommends for official control the European Pharmacopoeia methods to determine Niacin and Niacinamide in the feed additive.

For the determination of Niacin and Niacinamide in premixtures, the Applicants propose two methods based on ion-pair Reversed Phase High Performance Liquid Chromatography coupled to UV detector (RP-HPLC-UV), namely (1) the method developed by VDLUFA (Association of German Agricultural Analytical and Research Institutes) and (2) a single-laboratory validated and further verified method submitted by one of the Applicants. The VDLUFA method was ring-trial validated with a Niacin and Niacinamide content ranging from 3665 to 15540 mg/kg premixtures. The following performance characteristics were reported for Niacin and Niacinamide: (i) a relative standard deviation for repeatability (RSD_r) ranging from 3.49 to 7.58 %, and (ii) a relative standard deviation for reproducibility (RSD_R) ranging from 4.02 to 15.4 %.

The above mentioned single-laboratory validated and further verified method is also suitable for the determination Niacin and Niacinamide in feedingstuffs. The performance characteristics of this method were recalculated by the EURL using the experimental data provided by the Applicant, to obtain:

* For premixtures samples with contents ranging from 0.4 to 40 g/kg, for Niacin and Niacinamide:

- RSD_r ranging from 0.5 to 0.8 %;

- RSD_ip (intermediate precision) ranging from 4.9 to 6.7 %;

- R_Rec (recovery rate) ranging from 96.9 to 106.1 %, and

* For feedingstuffs samples with contents ranging from 8 to 800 mg/kg:

- RSD_r ranging from 4.1 to 5.4 %;

- RSD_ip ranging from 4.1 to 11.1 %;

- R_Rec ranging from 95.7 to 107.0 %, and

- LOD and LOQ of 2 and 8 mg/kg, respectively.

For the determination of Niacin and Niacinamide in water the Applicant provided data, upon request of the EURL, proving that the method for premixtures and feedingstuffs is also applicable when analysing Niacin and Niacinamide in water, without any sample preparation except a simple dilution.

Based on the above considerations and the performance characteristics presented, the EURL recommends for official control the single-laboratory validated and further verified method submitted by the Applicant, based on ion-pair Reversed Phase High Performance Liquid Chromatography coupled to UV detector (RP-HPLC-UV), to determine Niacin and Niacinamide in premixtures, feedingstuffs and water, within the concentration range covered by the experimental data. Additionally, the EURL recommends for official control the VDLUFA (Method Bd. III, 13.9.1) using RP-HPLC-UV, to determine Niacin and Niacinamide in premixtures.

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.

¹ FAD-2010-0112; ² FAD-2010-0263; ³ FAD-2010-0198; ⁴ FAD-2010-0265; ⁵ FAD-2010-0397

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2024/1194

For the determination of nicotinic acid in the feed additive: 

• Titration with sodium hydroxide; European Pharmacopoeia monograph 0459

For the determination of nicotinic acid in premixtures, compound feed and water:

• Ion-pair Reversed Phase High Performance Liquid Chromatography coupled to UV detector (RP-HPLC-UV)

For the determination of niacinamide (nicotinamide) in the feed additive:

• Titration with perchloric acid; European Pharmacopoeia monograph 0047

For the determination of niacinamide (nicotinamide) in premixtures, compound feed and water:

• Ion-pair Reversed Phase High Performance Liquid Chromatography coupled to UV detector (RP-HPLC-UV)