Analytical Sciences

Abstract − Analytical Sciences, 23(3), 357 (2007).

Application of Alkaline Phosphatases from Different Sources in Pharmaceutical and Clinical Analysis for the Determination of Their Cofactors; Zinc and Magnesium Ions
Svetlana V. MUGINOVA, Anna M. ZHAVORONKOVA, Alexei E. POLYAKOV, and Tatyana N. SHEKHOVTSOVA
Chemistry Department, M. V. Lomonosov Moscow State University, Leninskie Gory, Moscow 119992 GSP-2, Russia
Prospects of using different alkaline phosphatases bearing zinc and magnesium ions in their catalytic and allosteric sites, respectively, in pharmaceutical and clinical analysis were demonstrated. Also their application for the determination of zinc in insulin to control injection quality and magnesium in human urine for the diagnosis and treatment of magnesium deficiency was shown. The reaction of p-nitrophenyl phosphate hydrolysis was chosen as an indicator. The choice of appropriate alkaline phosphatase was substantiated, the influence of the nature of buffer solutions on the behavior of the enzyme-metal systems was studied, and the conditions of the indicator reaction proceeding in the presence of sample matrixes were optimized. Simple, rapid, sensitive, and selective enzymatic procedures for determining zinc and magnesium based on their inhibiting and activating effects on the catalytic activity of alkaline phosphatases from seal and chicken intestine, respectively, were developed.