A Micro Colorimetric Method for Determination of Blood Lactate Using Lactate Oxidase

概要

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An accurate enzymatic procedure to for the determination of blood lactate is explored. The hydrogen peroxide is produced from lactate which is oxidized by lactate oxidase extract from Pediococcus sp. It is oxidatively coupled with 4-aminoantipyrin and N-ethyl-N-(2-hydroxy-3-sulfopropyl)-m-toluidine sodium salt (TOOS) by peroxidase to yield a chromogen with an absorption maximum at 550 nm.<BR>As the procedure preparing protein-free supernatant, Somogyi method was employed, but with a exception that the order of adding the barium hydroxide soiution and zinc sulfate solution is reversed.<BR>The method requires only 25 μl of ear-lap blood or venous blood and a 15 min. incubation. The present method offers a measurement of high concentration of lactate up to 10 m mol/l blood.<BR>The results obtained by this method show good correlation with those obtained by the LDH-Hydrazin UV method and the LDH-GPT UV method (correlation coefficient is 0.99).<BR>The method has a highly specific that requires a smaller quantity of sample and a shorter time than those previously reported, and has excellent precision.
Japan Society of Clinical Chemistryの論文