Spectrophotometric and electrochemical determinations of L(+)-lactate in blood by use of lactate dehydrogenase from yeast.

Abstract

Lactate can be determined rapidly in blood by spectrophotometric and amperometric (enzyme electrode) procedures based on its oxidation by ferricyanide, the reaction being catalyzed with yeast L(+)-lactate dehydrogenase (cytochrome b2) (EC 1.1.2.3). In the photometric method lactate can be measured in a few minutes, but blood samples must first be deproteinized. In the amperometric procedure no treatment of blood is needed except ferricyanide addition. The enzyme electrode we used has a response time shorter than 1 min when its critical variables are optimized. Preliminary standardization is reduced to minimum operation, because electrode response is proportional to lactate concentration over a wide range (0.1 to 8.0 mol/liter) and many determinations can be done with little cost in enzyme. A simple electrical device ("two-electrode device") is described that is well suited for furture micro-cell construction. Lactate determinations on a series of normal blood samples show no deviation between results by these new methods and the usual ultraviolet spectrophotometric lactate tests.