Determination of Low Levels of GOT Activity by a Diazonium Salt Coupling and Ultraviolet Methods, using “System Olli 3000” Analyzer.

Abstract

In order to measure low levels of mitochondrial GOT (m-GOT) activity in eluates from DEAE-Sephadex A-50 minicolumn for the separate determination of serum GOT isoenzyme, a diazonium coupling procedure, modified from the method of Amador et al., and ultraviolet method for GOT determination were studied using System Olli 3000, a semiautomated computer-controled discrete type analyzer for end point and kinetic assay.
To increase the sensitivity of the Fast Violet B dye method, low volume of the concentrated buffered substrate relative to that of sample is used, the incubation time is prolonged to E0 min., and the coloured product is measured at a fixed time after the addition of colour reagent without use of a diluent. By these means, GOT activity as low as 0.2 Karmen units can be precisely determined with a linearity of up to 6 Karmen units by the instrument.
The photometer and the computational program developed for kinetic assay of the instrument exhibit several advantages for measuring GOT activity as low as 0.5 Karmen units by ultraviolet method with good precision.