1988 Fiscal Year Final Research Report Summary
Development of Automatic Method for Determining Enzyme Activities in Serum Using Bioreactor System
| Project/Area Number |
62870108
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| Research Category |
Grant-in-Aid for Developmental Scientific Research
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| Allocation Type | Single-year Grants |
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| Research Field |
Laboratory medicine
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| Research Institution | Kyoto University |
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Principal Investigator |
MURACHI Takashi Faculty of Medicine, Kyoto University, 医学部, 教授 (10089104)
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| Co-Investigator(Kenkyū-buntansha) |
TOTANI Masayuki Faculty of Medicine, Kyoto University, 医学部, 講師 (70163988)
TABATA Masayoshi College of Medical Technology, Kyoto University, 医療技術短期大学部, 助教授 (70115880)
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| Project Period (FY) |
1987 – 1988
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| Keywords | bioreactor / FIA / chemiluminometry / serum enzyme activity / lactate dehydrogenase (LDH) / 血清酵素活性 |
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| Research Abstract |
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.
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