Determination of Organic Phosphates in Rat Heart Muscle with an Organic Phosphates Analyzer

Abstract

The organic phosphates analyzer, originally developed by us based on the anion-exchange separation on an Asahipak NH2P-50 column and the postcolumn phosphomolybdic acid reaction, was improved for the analysis of biological materials. The anion-exchange separation was optimized to resolve several model phosphates by using a LiCI linear gradient elution. An acetonitrile pretreatment method was devised for simple and effective deproteinization and the extraction of organic phosphates in rat heart muscle. The analyzer was successfully applied to the determination of organic phosphates in rat heart muscle.

Fluorometric Determination of Phospholipids by High-Performance Liquid Chromatography with Postcolumn On-Line Hydrolysis under UV-Irradiation, Phosphomolybdic Acid and Thiamine–Thiochrome Reactions

“…There is, however, no general method for the HPLC determination of phospholipids, except for liquid chromatography with mass spectrometry (MS) 16 , MS coupled with ELSD 17,18 and flame photometric detection. 19 We also developed a postcolumn detector for organic phosphates and reported on its applications to watersoluble organic phosphates [20][21][22][23] , and on a selective separation of nucleosides 24 and organic phosphates 25 on a titania column. We previously described that some hydrophobic organic phosphates, like phospholipids, could be detected by using on-line hydrolysis under UV-irradiation on the basis of conversion into orthophosphate in a flow-injection system (FIA), in which a phosphomolybdic acid reaction and then a thiamine-thiochrome fluorescent reaction occurred.…”

A Novel Method for the On-Line Room-Temperature Hydrolysis of Hydrophobic Organic Phosphates Using a Cooperative Action of Sodium Dodecyl Sulfate, Acid and UV-Irradiation

“…Some methods have been reported for the on-line decomposition of organic phosphates using cobalt(III) oxidation 1 , peroxodisulfate digestion 2,3 , enzymatic dephosphorylation 4 , and oxidative decomposition with light 5,6 and microwaves. 7,8 We also reported on the on-line hydrolysis of organic phosphates by sulfuric acid 9 and its application [10][11][12] to their determination. The organic phosphates examined so far were all water-soluble.…”