Riboflavin from Bacillus subtilis CGMCC 7.449 in feed and water: in the additive Ph. Eur. monograph 0292 or HPLC-UV according to VDLUFA 13.9.1, in compound feed and water HPLC-FLD according to EN 14152
Commission Implementing Regulation (EU) 2025/1795, published on 10 September 2025, authorises until 29 September 2035 for all animal species two forms of vitamin B2 from fermentation with the genetically modified strain B. subtilis CGMCC 7.449: riboflavin of purity ≥ 98 % (3a825iii, water ≤ 1.5 %) and a preparation with ≥ 80 % riboflavin (3a825vi, water ≤ 3 %). Both forms may be used in water for drinking.
- Methods in the annex: 98 % additive — European Pharmacopoeia monograph 0292 or HPLC-UV (VDLUFA Bd. III, 13.9.1); premixtures and the 80 % preparation — HPLC-UV (VDLUFA); compound feed and water — HPLC with fluorescence detection (EN 14152), result as total vitamin B2.
- EFSA data for the 98 % form: in six batches 98.6–98.8 % riboflavin in dry matter (applicant’s specification 97–103 %, in line with Ph. Eur.), impurity A (lumiflavin) 0.0025–0.0084 % against a limit of 0.025 %, total impurities 0.2–0.4 % (limit 0.5 %), specific optical rotation −128…−131° (specification −115…−135°), absorbance ratios A373/A267 0.32–0.33 and A444/A267 0.38. The 80 % preparation (with maltodextrin): 83.8–84.2 % riboflavin.
- The production strain carries acquired antimicrobial resistance genes, but no viable cells or spores were detected in the products; strain DNA was tested only in the 98 % form — no such information for the 80 % preparation. In water (10 mg/L, 25 °C, 48 h) no significant losses; homogeneity in water — CV 8.5–8.7 %.
- Users: the additive is not a skin or eye irritant but is a skin sensitiser (OECD TG 442E and 442D), and is therefore also considered a respiratory sensitiser; dusting potential 451–507 mg/m³ for the 98 % form and 22–42 mg/m³ for the 80 % form. The act requires respiratory and skin protection if procedures do not eliminate the risks.