The Effects of Storage Conditions and Homogenisation Buffers on the Measurement of SOD, CAT and ADA Enzyme Activities in Cattle Liver

Abstract

ABSTRACTBackgroundAccurate measurement of enzyme activities is very important in studies to evaluate enzymatic parameters. While performing these measurements, many factors can affect the results, including the method of obtaining the tissues, the conditions under which they are stored until analysis, and the methods of determining enzyme activity.ObjectivesThis study aimed to investigate the effect of different storage conditions (time and temperature) and different homogenisation buffers (PBS or KCl) on the enzymatic activity of superoxide dismutase (SOD), catalase (CAT), and adenosine deaminase (ADA) in homogenised cattle liver.MethodsFresh liver tissue samples were obtained from the slaughterhouse and homogenised in different homogenisation buffers. Supernatants from each sample were divided into three groups according to the experimental design of the study. SOD, CAT and ADA enzyme activities in homogenised tissues were evaluated.ResultsOur data revealed that SOD, CAT and ADA activities did not differ significantly between PBS and KCl groups or between ‐20°C and ‐80°C freezing temperatures. However, our results showed that SOD levels decreased over time in both groups, CAT levels demonstrated a significant decrease from Month 0 to Month 3 and ADA levels decreased significantly over time.ConclusionsThe results indicated that storage time had a significant effect on enzyme activity changes, but the effect of storage temperature and homogenisation buffer was generally limited. These results may support the measurement of enzymatic activity in liver homogenate immediately and, if necessary, after freezing for the shortest possible time.