High-performance liquid chromatography determination of enzyme activities in the presence of small amounts of product

摘要: Enzymes can be assayed by HPLC calculating the amount of substrate(s) left over, or product formed, through peak area ratios with a suitable internal standard. However, sometimes substrates used are contaminated small amounts products and this lead to errors in determination enzyme activity. A method for test such enzymes, which prevents eventual errors, uses ratio substrate/product at time zero as standard kinetics followed aid simple mathematical equation. This approach was applied activities papain, urokinase, NAD glycohydrolase, pyruvate kinase samples it compared data obtained method, giving reproducible results all cases.