Separation of Thiamin, Thiamin Antagonists and Their Phosphate Esters by High-Performance Liquid Chromatography

Abstract
Methods are presented for the separation of the phosphate esters of thiamine, oxythiamine or pyrithiamine and the separation of thiamine and oxythiamine by high-performance liquid chromatography. Detection by means of a UV monitor and fluorometer with a post-column oxidation system is described which allows for selective detection of thiochrome, pyrichrome and their respective phosphate esters. Anion exchange and ion-pair reverse phase chromatography are the chromatographic modes of separation.