Verfahren zur Bestimmung der alkalischen Phosphatase-Aktivität

Abstract

The activity of alk. phosphatase is detd. by incubation of a sample with NADP (I) in a buffer medium with subsequent spectrophotometric recording of the hydrolysis product in an enzymic regeneration system of a cofactor-contg. alk. dehydrogenase and two substrates, one of which is EtOH. The sensitivity of the detn. is increased and a rapid and simplified detn. is ensured by incubation of the sample and recording of the hydrolysis product catalyzed by alk. phosphatase in a pH 8,5 - 9,1 tris-HCl buffer; By using I in reduced form at 10-7 - 10-4 M; by using horse liver alk. dehydrogenase at 5 x 10-7 - 10-5 M the cofactor regeneration system; by using p-nitroso-N,N'-dimethylamine at 5 x 10-6 - 3 x 10-4 M as th 2nd substrate; and by adding salicyclic acid amide to 2 x 10-4 - 5 x 10-7 M and Na orthophosphate to 0,05 M to the system.