Assay Optimization of Lactate Levels in Athletes Using Nanophotometry Methods

Abstract

Lactate levels are the main parameter used in sports as a marker of fatigue. In general, the measurement of lactate levels in research uses the rapid test diagnostic test method. Biochemically there will be a difference between rapid test measurements with enzymatic methods. But there are many factors to consider in enzymatic testing. The purpose of this study is to optimize the substrate volume and total reaction volume in enzymatic measurement of lactate levels using a nanophotometer. Lactate can be detected in biological samples such as serum, red blood cells, cell culture, and fermentation media. In this study serum was used as a sample obtained from a university student who carried out swimming activities as far as 200m. Then the serum is used to measure lactate levels using the Lactate Assay Kit (MAK064, Sigma Aldrich). Lactate levels were read calorimetric at a wavelength of 570nm with a nanophotometer. Optimization results show the optimum substrate volume is 1.5 ul with 11 ul buffer with the final reaction volume of 25 ul. Standard curves showed regression values (R2) 0,9909, and lactate levels in2,848nmol/µl samples.