Abstract
The great usefulness of the previously reported "stopped-flow time difference analysis" technique for improving enzymatic analyses based on dehydrogenase-catalyzed reactions is demonstrated by combining the technique with the conventional enzymatic method for pyruvate determination. Reagent economy was accomplished by using a micro-stopped-flow apparatus designed by one of the authors. The running speed and sensitivity were improved by about 100 and 7 times the figures for the conventional enzymatic method, respectively. Furthermore, as procedural blanks were unnecessary, a simpler method could be developed. The interference by several other keto acids was easily eliminated by analyzing the reaction curve, and the apparent first-order rate constant proved to be useful for qualitative analysis.
