Stability of NADPH: Effect of various factors on the kinetics of degradation

Abstract

Seeking to minimize degradation of NADPH during storage, reagent preparation, and assays, we investigated the effects of pH, temperature, and ionic strength as well as the effects of phosphate and acetate. NADH was also included for comparison. Our results indicate that the rate of degradation of NADPH is proportional most importantly to temperature and concentrations of hydronium ion, but also to concentrations of phosphate and acetate. The degradation rate decreased with increasing ionic strength at neutral pH, but increased slightly at lower pH. NADPH generally is less stable than NADH under the same conditions. The reaction orders with respect to hydronium ion and anions were near 1 for NADH degradation reactions, about 0.5 for NADPH. Rate constants for NADH and NADPH differed more at higher pH and lower phosphate and acetate concentrations.