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AOAC First Action Method 2011.10, Vitamin B12 in Infant and Pediatric Formulas and Adult Nutritionals, was collaboratively studied. This method uses a pH 4.5 sodium acetate buffer and potassium cyanide at 105°C to extract and convert all biologically active forms of vitamin B12 present to cyanocobalamin; octylsilyl (C8) or C18 SPE cartridges to purify and concentrate cyanocobalamin; a combination of size-exclusion and RPLC to isolate cyanocobalamin; and visible absorbance at 550 nm to detect and quantitate cyanocobalamin in infant, pediatric, and adult nutritionals with vitamin B12 concentrations greater than 0.025 μg/100 g ready-to-feed (RTF) liquid. During this collaborative study, nine to 11 laboratories from eight different countries analyzed blind duplicates of 12 infant, pediatric, and adult nutritional formulas. Per the AOAC Expert Review Panel (ERP) on Stakeholder Panel on Infant Formula and Adult Nutritionals (SPIFAN) Nutrient Methods the method demonstrated acceptable repeatability and reproducibility and met SPIFAN Standard Method Performance Requirements (SMPRs®) for the majority of product matrixes analyzed. Vitamin B12 SPIFAN SMPRs for repeatability were ≤15% RSD at vitamin B12 concentrations of 0.01 μg/100 g RTF liquid and ≤7% RSD at vitamin B12 concentrations of 0.2-5.0 μg/100 g RTF liquid. Vitamin B12 SPIFAN SMPRs for reproducibility were ≤11% RSD in products with vitamin B12 concentrations ranging from 0.3 to 5.0 μg/100 g RTF liquid. During this collaborative study, the RSDr ranged from 2.98 to 9.77%, and the RSDR ranged from 3.54 to 19.5%. During previous single-laboratory validation studies, the method LOQ was estimated to be 0.025 μg/100 g RTF liquid. [ABSTRACT FROM AUTHOR]

The rate of vitamin B12 loss was 3 X greater in heated, chocolate-flavoured milk than in unflavoured milk. Studies of B12 stability in the presence of cocoa powder components were performed to identify the reactive agents. Cocoa polyphenols with a strong capacity both for reduction and for peroxide generation accelerated B12 destruction as much as 20-fold vs. polyphenols without both capacities. Polyphenols with both capacities include the cocoa polyphenol (+)-catechin and oligomers thereof, but also the structurally related polyphenols gallic acid, caffeic acid and (−)-epigallocatechin gallate. The demonstrated physical affinity of cocoa powder for B12 was a significant factor in the destructive process. B12 may exhibit decreased stability in heated, neutral p H, polyphenol-fortified foods. [ABSTRACT FROM AUTHOR]