气相色谱-质谱法测定人血浆中稳定同位素取代的组氨酸

Determination of stable isotopically substituted histidine in human plasma by gas chromatography-mass spectrometry.

作者信息

Furuta T, Kasuya Y, Shibasaki H, Baba S

出版信息

J Chromatogr. 1987 Jan 23;413:1-7. doi: 10.1016/0378-4347(87)80207-4.

Abstract

A gas chromatographic-electron-impact mass spectrometric method for the determination of stable isotopically substituted histidine in human plasma has been developed. Histidine was derivatized to alpha N-trifluoroacetyl-imN-carbethoxyhistidine n-butyl ester (TCB derivative) by a three-step reaction: an initial esterification by 3 M n-butanolic hydrochloric acid, followed by trifluoroacetylation with trifluoroacetic anhydride and then ethoxycarbonylation with diethyl pyrocarbonate. Quantitation was carried out by selected-ion monitoring on the molecular ions (m/z 379, 383 and 385) of the respective TCB derivatives of histidine, [1-15N,5,beta,beta-2H3]histidine (histidine-[M + 4]) and [1,3-15N2,5,alpha,beta,beta-2H4]histidine (histidine-[M + 6]). The sensitivity, specificity, precision and accuracy of the method were demonstrated to be satisfactory for application to a pharmacokinetic study of histidine after administration of a trace amount of stable isotopically substituted histidine (histidine-[M + 4]) in humans.

摘要

已开发出一种气相色谱 - 电子轰击质谱法，用于测定人血浆中稳定同位素取代的组氨酸。组氨酸通过三步反应衍生化为α - N - 三氟乙酰基 - imN - 乙氧羰基组氨酸正丁酯（TCB衍生物）：首先用3M正丁醇盐酸进行酯化反应，接着用三氟乙酸酐进行三氟乙酰化反应，然后用焦碳酸二乙酯进行乙氧羰基化反应。通过对组氨酸、[1 - 15N,5,β,β - 2H3]组氨酸（组氨酸 - [M + 4]）和[1,3 - 15N2,5,α,β,β - 2H4]组氨酸（组氨酸 - [M + 6]）各自的TCB衍生物的分子离子（m/z 379、383和385）进行选择离子监测来进行定量分析。该方法的灵敏度、特异性、精密度和准确度已被证明对于在人体中施用痕量稳定同位素取代的组氨酸（组氨酸 - [M + 4]）后进行组氨酸的药代动力学研究而言是令人满意的。