The use of a common visible indicator reaction in the development and application of several sensitive analytical chemistry methods.

摘要

A sensitive visible indicator system was used in the development of several assays for analytes of clinical interest. These assays employed a modified Trinder reaction in which 2-hydroxy-3,5-dichlorobenzenesulfonate was substituted for phenol, thereby increasing the sensitivity of the chromogen several-fold and thus facilitating the development of new and better quantitative methods for several clinical analytes.;Two kinetic, colorimetric assays for the quantitation of creatine kinase activity in serum were developed. The first assay employed a creatinase-sarcosine oxidase coupling mechanism, and thus, was free from interference by endogenous adenylate kinase. The second assay employed a glycerol kinase-glycerophosphate oxidase coupling mechanism. The latter assay was adapted to the measurement of creatine kinase isoenzymes following electrophoretic separation in agarose gel.;Colorimetric end-point assays were developed for the measurement of ATP and glycerol in millimole per liter concentrations using modifications of the glycerol kinase-glycerophosphate oxidase reaction sequence.;A kinetic, colorimetric method was developed for the measurement of serum magnesium ion concentration. This method utilizes the magnesium-dependent enzyme glycerol kinase, whose activity in the colorimetric reagent system is proportional to the serum magnesium ion concentration.;Finally, a novel reagent mechanism was developed which allows the coupling of dehydrogenase enzyme systems to the peroxidase colorimetric indicator reaction. The effectiveness of the coupling mechanism is illustrated by the development of quantitative end-point assays for serum glucose and magnesium ion.