Butylated hydroxyanisole - 1b320

In the current application authorisation is sought for Butylated Hydroxy Anisole, E320 (BHA) under Article 10, category/functional group 1(b) 'technological additives'/'antioxidants', according to the classification system of Annex I of Regulation (EC) No 1831/2003. BHA is already authorised as feed additive under Commission Directive 70/524/EEC. 

According to the Applicant BHA is a white or slightly yellow waxy solid with a minimum purity of 98.5% BHA, containing at least 85% of 3-tert-butyl-4-hydroxyanisole (3-BHA) and 2-tert-butyl-4-hydroxyanisole (2-BHA) and a maximum of 0.2 % of hydroquinone. Specifically, authorisation is sought for the use of the feed additive for all animal species and categories. The feed additive is intended to be mixed in premixtures or added directly in complete feedingstuffs. Furthermore, the Applicant proposed a maximum level in the daily ration of 150 mg/kg for BHA alone or for the sum of BHA with Butylhydroxytoluene (BHT, E321) and/or Ethoxyquin (E324).

For the determination of BHA in the feed additive, the Applicant submitted the Food Chemical Codex 7 (FCC) method based on Gas Chromatography coupled to Flame Ionization Detection (GC-FID). Standard deviations for repeatability (RSD_r) of 2.0%, for 3-BHA, and 6.0%, for 2-BHA, were reported.

Based on the performance characteristics presented, the EURL recommends for official control the FCC 7 method based on GC-FID to determine BHA in the feed additive.

For the determination of BHA in premixtures and feedingstuffs the Applicant submitted a single laboratory validated and further verified multi-analyte method, based on Reversed Phase High Performance Liquid Chromatography coupled with UltraViolet-Diode-Array Detection (RP-HPLC-UV-DAD). The following correspondent performance characteristics were reported for concentrations in premixtures ranging from 5 to 120 g/kg and concentrations in feedingstuffs ranging from 35 to 226 mg/kg:

- a RSD_r ranging from 0.6 to 3.9%;

- a standard deviation for intermediate precision (RSD_ip) ranging from 2.6 to 6.4%;

- a recovery rate (R_Rec) ranging from 90.5 to 114%; and

- a limit of quantification (LOQ) below 35 mg/kg.

Based on the performance characteristics presented, the EURL recommends for official control, the single laboratory validated and further verified RP-HPLC-UV-DAD method, submitted by the Applicant, to determine BHA in premixtures and feedingstuffs.

According to the Applicant the above mentioned multi-analyte technique, submitted for the determination of BHA in premixtures and feedingstuffs, allows the quantification of other synthetic antioxidants such as BHT and Ethoxyquin (in premixtures only). Furthermore, the EURL identified the ring trial validated method by the Association of Official Analytical Chemists (AOAC 996.13 – "Ethoxyquin in feeds") based on isocratic RP-HPLC system coupled with fluorescence detection (RP-HPLC-FD). The following precisions (repeatability and reproducibility) were reported: ranging from 1.4 to 4.6 for BHT in premixtures and feedingstuffs and ranging from 2.1 to 5.4 and 4.5 to 29% for Ethoxyquin in premixtures and feedingstuffs respectively.

Based on the performance characteristics presented, the EURL considers suitable for the quantification of BHT and Ethoxyquin:

- the single laboratory validated and further verified RP-HPLC-UV-DAD method, submitted by the Applicant, for BHT in premixtures and feedingstuffs and Ethoxyquin in premixture only, and

- the ring trial validated RP-HPLC-FD method characterised by the "Association of Official Analytical Chemists" (AOAC 996.13)  for Ethoxyquin in 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) is not considered necessary.

Recommended Method

as of:

- Commission Implementing Regulation (EU) 2020/1399

- Commission Implementing Regulation (EU) 2022/654

For the quantification of BHA in the feed additive: 

 Gas Chromatography coupled to Flame Ionization Detection (GC-FID) (FCC7 method) 

For the quantification of BHA in premixtures and feedingstuffs: 

- Reversed Phase High Performance Liquid Chromatography coupled to UltraViolet - Diode-Array Detection (RP-HPLC-UV- DAD, 285 nm)