Agroscope Liebefeld-Posieux Research Station ALP, Schwarzenburgstrasse 161, CH-3003 Berne, Switzerland.
Meat Sci. 2012 Feb;90(2):279-83. doi: 10.1016/j.meatsci.2011.07.009. Epub 2011 Jul 30.
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 50mM 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>0.99. 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.
建立了一种通过反相高效液相色谱和紫外检测定量测定肉制品中维生素 B12 的方法，并与参考方法（微生物测定法，MBA）进行了比较。维生素 B12 用 50mM 乙酸钠缓冲液在氰化钠存在下提取。为了定量测定总维生素 B12，需要通过胃蛋白酶处理释放与蛋白质结合的维生素 B12。氰钴胺素在免疫亲和柱上进行纯化和富集后，被检测为总维生素 B12。用五种浓度的维生素 B12 进行校准，具有良好的线性关系，回归系数 r2>0.99。该方法在三个不同浓度水平（5-15ng/g）下进行了验证，香肠的回收率在 80-108%之间，相对标准偏差在 1.50-7.26%（n=6）之间较低。检测限为 2ng/g。用所建立的方法测定的 50 种肉类产品的维生素 B12 水平与 MBA 测定的水平相似或显著降低。
