乳酸脱氢酶反应器用于流动注射分析测定人体液中的L-乳酸

Flow Injection Analysis for L-Lactate in Human Fluid With Immobilized Dehydrogenase Reactor

本文用[3-(2-氨乙基)氨丙基]三乙氧基硅烷试剂胺化一定孔径大小的载体,用戊二醛作偶联剂,将乳酸脱氢酶固定于其上,装入柱内,联结于流动体系中。L-乳酸的浓度在0.1～4.4mmol/L呈线性,检测限为0.4nmol(s/N=4),相对标准偏差≤1.2％,回收率在96％～103％。每小时进样35次。测定了固定化LDH的表观米氏常数及人血清和尿中的L-乳酸。

The L-lactate dehydrogenase (L-LDH) is immobilized by reaction with glutaraldehyde onto the surface of [3-amino-2-(2-aminoethyl) propyl]-aminopropyl] triethoxysilane controlled porous glass beads and packed into a column in the flow system. The relative standard deviation is ≤1.2％ with recovery of 96％～103％ for the L-lactate concentration range of 0.1～4.4 mmol/L, detection limit of 0.4 nmol(S/N=4) and sampling frequency of 35 samples/h, The apparent Michaelis constant of immobilized LDH and L-lactate in human serum and urine are determined.