Immobilization of Enzymes and their Applications in "Flow Injection Analysis" For the Determination of substrates of Diagnostic Importance

Abstract

This project was initiated on nov.1th, 1987 and has been concluded on October.31th, 1990. Five reports including 2 annual and 3 semi-annual reports have already been submitted in the past. This final report is the sort of collection from all previously submitted reports. The achievement made are summarized below: these include, a) Construction of a flow injection system equipped with an electrochemical as well as spectrophotometric detector. b) Standardization and calibration of the system with standard solution of hydrogen peroxide. c) Selection of suitable support for the immobilization of enzymes and the preparation of a new support, a phenolic resin and evaluation study. d) Application of the system to the analysis of glucose in blood. e) Determination of total and free cholesterol in blood using immobilized enzymes in the establishment flow injection system. f) Establishment of automated procedure for uric acid in blood using urate oxidase. g) Similar procedure were established for xanthine, hypoxanthine and xanthine oxidase. h) In connection with cholesterol estimated attempts are still in progress to establish a procedure for LDL-cholesterol estimation. i) Novel immobilized enzyme procedures were established for phospholipids which in fact makes the heart of this final report. j) Development of immobilized enzymes based procedure for lipid bound sialic acid in serum was attempted a couple of times but no success was made. The reason being the very heat sensitive nature of enzymes, aldolase and very N-acetyl neuraminidase which were received in the laboratory from the commercial firm twice in the denatured form. The only alternative now would be to isolate and purify these enzymes in the lab and then use them.