Development of Automatic Method for Determining Enzyme Activities in Serum Using Bioreactor System

血清中的酶活性已被广泛地确定在临床分析中使用自动分析仪。最近，对血清中非蛋白质有机化合物的自动分析通过生物反应器在列形式中不流动的wnzyme已被用于实际使用。为了应用生物反应器的目的，以确定酶活性在血清中的作用，并增加其使用范围，我们研究了如下内容，并获得了大量的结果。（1）对酶反应的孵化系统的研究。由于系统将反应线圈用作酶的反应，即流动注射分析（FIA）中的旁路，因此系统既简单又容易去除样本空白，我们选择系统。（2）使用生物反应器血清对酶活性的自动分析的发展。1.血清中的乳酸脱氢酶（LDH）活性。LDH反应是在反应线圈的基底上放置吡啶酯，作为旁路在FIA系统中产生的，而LDH反应是通过乳酸氧化酶生物反应器氧化化为吡啶酯和羟基过氧化物。氢过氧化物的生产是由化学物质法确定的。内源性乳酸是由乳酸氧化酶和催化剂预列清除的。该结果显示，日立模型726自动分析仪提供了出色的和相关的满意度。目前的方法并不需要任何空白的样本。血清中淀粉酶活性的决定。麦芽糖被用作淀粉酶反应的基础。由淀粉酶反应产生的麦芽糖被麦芽糖磷酸酶和幽门螺杆菌氧化酶生物反应器催化到羟基过氧化物。葡萄糖激酶柱被用来去除血清中的内源性葡萄糖。取得的结果很好。

Enzyme activities in serum have been determined widely using an automated analyzer in clinical analysis. Recently, the automated analysis of nonprotein organic compounds in serum by a bioreactor with immobilized wnzyme in column form has been put to practical use. For the purpose of applying the bioreactor to the determination of enzyme activities in serum and increasing the range of its use, we studied as follows, and obtained a lot of results.(1) Study of the incubation system for the reaction of enzyme. Since the system putting a reaction coil for the reaction of enzyme as a bypath in the flow injection analysis (FIA) system was simple and easier to remove sample blanks, we chose the system.(2) Development of the automatic analysis of enzyme activities in serum using a bioreactor. 1. Determination of lactate dehydrogenase (LDH) activities in serum. LDH reaction was taken place using pyruvate as the substrate in the reaction coil put as a bypath in the FIA system, and lactate produced by LDH reaction was oxidized to pyruvate and hydrogen peroxide by the lactate oxidase bioreactor. Hydrogen peroxide produced was determined by chemiluminescence method. Endogenous lactate was removed by lactate oxidase・catalase precolumn. The results obtained were excellent, and correlated satisfactorily with those by a Hitachi Model 726 automatic analyzer. The present method did not need any sample blank.2. Determination of amylase activity in serum. Maltopentaose was used as the substrate for amylase reaction. Maltose produced by amylase reaction was catalyzed to hydrogen peroxide by the maltose phosphorylase・pyranose oxidase bioreactor. Glucokinase column was used to remove endogenous glucose in serum. The results obtained was good.