HPLC Analysis of Vitamin B_6 Derivatives

为了测定天然产物中普遍存在的维生素B_6衍生物的含量，本文研究了本实验室新开发的等度反相高效液相色谱法（HPLC）对血、尿或某些哺乳动物器官样品和复杂食品样品的分析适用性。测定数据为维生素B_6的推荐膳食供给量（RDA）提供了标准。对于项目1），我们建立的高效液相色谱法为分析维生素B_6衍生物在某些器官以及血、尿中的浓度变化提供了有力的手段。特别是维生素B_6衍生物的主要成分5 '-磷酸吡哆醛（PLP），经KCN处理转化为5'-磷酸吡哆酸（PIC-P）后，可有效地进行分析，而高氯酸萃取后，可用于某些食品和食品原料中总维生素B_6含量的分析。估计值刚好低于通过使用酿酒酵母ATCC 9080的微生物测定以及通过来自HCl水解后的HPLC方法的数据获得的值。从复杂的食品基质中提取维生素B_6衍生物的方法必须重新进行精确的研究，以使该方法适用于食品分析。

In order to determine the contents of vitamin B_6 derivatives, which are ubiquitously found in natural products, isocratic reversed-phase HPLC method, newly developed in our laboratory, was examined whether or not the method was applicable to the analysis from 1) blood, urine or some mammalian organs and 2) complex food samples. The determined data offer the standard for the recommended dietary allowance (RDA) for vitamin B_6. For the project 1) , our HPLC method gave a powerful means to analyze the concentration changes of vitamin B_6 derivatives in some organs as well as in blood, urine. Especially, pyridoxal 5'-phosphate (PLP) , a main component of vitamin B_6 derivatives, was effectively analyzed after the conversion of it to pyridoxic acid 5'-phosphate (PIC-P) by means of KCN treatment.On the other hand, the analysis of total vitamin B_6 contents in some foods and food materials when analyzed after perchloric acid extraction, was estimated just lower than the values which were obtained by a microbiological assay using Saccharomyces cerevisiae ATCC 9080 as well as by data from HPLC method after HCl-hydrolysis.In summary, the extraction procedure of vitamin B_6 derivatives from complex food matrices have to be reexamined precisely to make the method applicable to the food analyzes.