A quantitative method for the determination of Vitamin B12 in meat products by RP-HPLC and UV detection
was developed and compared to the reference method (microbiological assay, MBA). Vitamin B12 was
extracted with 50 mM sodium acetate buffer in the presence of sodium cyanide. For the quantification of total
Vitamin B12, it was necessary to release protein-bound Vitamin B12 by pepsin treatment. Cyanocobalamin
was detected as total Vitamin B12 after purification and enrichment on an immunoaffinity column. The
calibration with five concentrations of Vitamin B12 was linear with a regression coefficient r2 N0.99. The
method was validated at three different concentration levels (5–15 ng/g) with salami showing good recovery
rates between 80 and 108% and low relative standard deviations between 1.50 and 7.26% (n=6). The
detection limit was found to be 2 ng/g. The Vitamin B12 levels of 50 meat products measured by the
developed procedure were similar or significantly lower than those determined by the MBA.