Poon H C, Schmidt B M
Children's Psychiatric Research Institute, London, Ontario, Canada.
Clin Biochem. 1991 Apr;24(2):149-52. doi: 10.1016/0009-9120(91)90466-r.
An enzymatic fluorometric assay to quantitate plasma pyridoxal 5'-phosphate (PLP) is described. PLP is preincubated for 30 min with purified tyrosine decarboxylase apoenzyme (TDA) in acetate buffer and is then incubated with L-tyrosine for 60 min. The decarboxylated metabolite, tyramine, is extracted into ethyl acetate, air dried, dissolved in borate buffer and reacted with fluorescamine to form a fluorophor (395/475 nm). Recovery of exogenous PLP was 83%. After log-normal transformation of 129 plasma PLP levels obtained from 3 groups of subjects (in-patients, out-patients, staff), a normal distribution curve was obtained yielding a mean of 83 nmol/L. With 95% confidence level, the reference limits were 32-170 nmol/L. There were no age or sex differences in mean plasma PLP levels between any of the 3 groups of subjects studied.
本文描述了一种用于定量血浆磷酸吡哆醛（PLP）的酶荧光测定法。PLP在醋酸盐缓冲液中与纯化的酪氨酸脱羧酶脱辅酶（TDA）预孵育30分钟，然后与L-酪氨酸孵育60分钟。脱羧代谢产物酪胺被萃取到乙酸乙酯中，空气干燥后，溶解在硼酸盐缓冲液中，并与荧光胺反应形成荧光团（激发波长395nm/发射波长475nm）。外源性PLP的回收率为83%。对从三组受试者（住院患者、门诊患者、工作人员）获得的129份血浆PLP水平进行对数正态转换后，得到一条正态分布曲线，其平均值为83nmol/L。在95%置信水平下，参考范围为32 - 170nmol/L。在所研究的三组受试者中，任何两组之间的血浆PLP平均水平均无年龄或性别差异。
