Novel phosphoric ester and a method for quantitative analysis of an alkaline phosphatase using the same as a substrate

摘要

PURPOSE:To determine the titled substance with a substrate having the improved detecting sensitivity and operating ratio without problems in synthesis and purification, by using a novel mono-4-arylazoarylphosphoric ester as a substrate. CONSTITUTION:An aromatic amine is diazotized and coupled with a phenolic derivative to synthesize an azo dye, which is then reacted with a large excess of phosphorus oxychloride in the presence of pyridine to afford a phosphorodichloridate. The resultant phosphorodichloridate is then reacted with water to give a phosphoric monoester. Sodium hydroxide, etc. is added to the phosphoric monoester, and crystals are deposited with ethanol to afford a mono-4-arylazoarylphosphoric ester of the formula (M is H or alkali; A is phenyl, naphthyl, etc.; B is p-phenylene, etc.). The resultant compound is then dissolved in a buffer solution, and a substance to be tested is added to the solution as a substrate solution to measure the formed azo dye concentration with a spectrophotometer.