Determination of phosphodiesterase activity in the presence of phosphomonoesterase using bis-p-nitrophenyl phosphate.

In the presence of phosphomonoesterase contaminations the use of bis-p-nitrophenyl phosphate to measure phosphodiesterase activity gives inconclusive values because one of the products of the phosphodiesterase or nuclease reaction becomes a substrate of the contaminating enzyme. A direct determination of the hydrolyzed phosphodiesterase substrate in the UV range is possible at the isosbestic points of the transformation of the phosphomonoesterase substrate.