Techniques for detecting enzymes in high-performance liquid chromatography.
Techniques are described for the automated detection of a series of enzymes in a high-performance liquid chromatographic system. Detection was achieved by either a direct or a coupled enzyme assay using photometric detectors. In direct detection the immediate enzymatic product was monitored. Coupled enzyme assays required additional enzyme(s) to convert the product of the primary enzyme reaction into a more easily detectable form. The efficiency of both free and immobilized coupling enzyme(s) was evaluated. The detector sensitivity could be increased three-fold by increasing the reaction temperature. This system is particularly suitable for isoenzyme profiling in biological materials.
D000818:Animals; D001322:Autoanalysis; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003402:Creatine Kinase / Q000097:blood; D004798:Enzymes / Q000097:blood*; D004800:Enzymes, Immobilized; D005954:Glucosephosphate Dehydrogenase / Q000097:blood; D006593:Hexokinase / Q000097:blood; D006801:Humans; D007527:Isoenzymes / Q000097:blood; D007770:L-Lactate Dehydrogenase / Q000097:blood; D008099:Liver / Q000201:enzymology; D008297:Male; D051381:Rats; D013737:Testis / Q000201:enzymology
D004798:Enzymes; D004800:Enzymes, Immobilized; D007527:Isoenzymes; D007770:L-Lactate Dehydrogenase; D005954:Glucosephosphate Dehydrogenase; D006593:Hexokinase; D003402:Creatine Kinase