Determination of Vitamin B12 in Infant Formula and Adult Nutritionals by Liquid Chromatography/UV Detection with Immunoaffinity Extraction: First Action 2011.08

Abstract

Abstract At the “Standards Development and International Harmonization: AOAC INTERNATIONAL Mid-Year Meeting,” on June 29, 2011, an Expert Review Panel agreed that the method “Determination of Vitamin B12 in Infant Formulas and Adult Nutritionals by Liquid Chromatography/UV Detection with Immunoaffinity Extraction” be adopted AOAC Official First Action status. The method is applicable for the determination of vitamin B12, which includes added cyanocobalamin and natural forms, making it applicable to both fortified and nonfortified products. Vitamin B12 is extracted from the sample in sodium acetate buffer in the presence of sodium cyanide (100°C, 30 min). After purification and concentration with an immunoaffinity column, vitamin B12 is determined by LC with UV detection (361 nm). A single-laboratory validation study was conducted on a range of products, including milk- and soy-based infant formulas, cereals, cocoa beverages, health care products, and polyvitamin premixes. The method demonstrated linear response over a large range of concentrations, recovery rates of 100.8 ± 7.5% (average ± SD), repeatability RSD (RSDr) of 2.1%, and intermediate reproducibility (RSDiR) of 4.3%. LOD and LOQ values were 0.10 and 0.30 μg/100 g, respectively, and correlation with the reference microbiological assay was good (R2 = 0.9442). The results of the study were published in J. AOAC Int. 91, 786–793 (2008). The performance characteristics of the method met the standard method performance requirements set forth by the Stakeholder Panel on Infant Formula and Adult Nutritionals; thus, the method was determined to be appropriate for First Action status.