Singkamani R, Worthington D J, Thurnham D I, Whitehead T P
Ann Clin Biochem. 1986 May;23 ( Pt 3):317-24. doi: 10.1177/000456328602300312.
The development of a simple enzymatic method for the determination of blood and plasma pyridoxal 5'-phosphate (PLP) using pig heart apo-aspartate transaminase (apo-AST) is described. The technique requires three steps: sample extraction using perchloric acid, a binding step in which PLP in the sample extract is attached to the apo-AST and the enzymatic assay of the reconstituted holo-AST. PLP extracts were analysed with and without a known concentration of added PLP to correct for variation in recovery between different specimens. Procedures are outlined for manual and automatic analysis of the PLP extracts. Using the KONE Clinical Analyser after the extraction step, it is possible to measure enzyme activity in 100 specimens (i.e. 400 tubes) in a 5 h period. Results are shown from 185 healthy women aged 20-45 years, in which plasma PLP concentrations ranged from 5 to 165 nmol/L, and 142 men and 56 women in government service aged 17-64 years, whose plasma PLP ranged from 8 to 169 nmol/L. Values less than 20 nmol/L are believed to indicate vitamin B6 deficiency and the method is able to measure 5 nmol/L.
本文描述了一种使用猪心脱辅基天冬氨酸转氨酶（apo-AST）测定血液和血浆中5'-磷酸吡哆醛（PLP）的简单酶法。该技术需要三个步骤：用高氯酸进行样品提取，样品提取物中的PLP与apo-AST结合的步骤，以及重组全酶AST的酶促测定。对添加已知浓度PLP和未添加PLP的PLP提取物进行分析，以校正不同标本间回收率的差异。概述了PLP提取物手动和自动分析的程序。在提取步骤后使用KONE临床分析仪，5小时内可测定100个标本（即400管）的酶活性。给出了185名年龄在20至45岁的健康女性的结果，其血浆PLP浓度范围为5至165 nmol/L，以及142名年龄在17至64岁的公职男性和56名公职女性的结果，其血浆PLP范围为8至169 nmol/L。低于20 nmol/L的值被认为表明维生素B6缺乏，该方法能够测定5 nmol/L。
