Highly Sensitive and Simple Method for Determination of Free 3-Methoxy-4-Hydroxyphenylglycol in Plasma by High-Performance Liquid Chromatography Using a Sep-Pak Alumina B Cartridge

Abstract

A novel sample clean-up procedure for the determination of free 3-methoxy-4-hydroxyphenylglycol (MHPG) in plasma in described. MHPG was purified with Sep-Pak alumina B cartridge followed by ethyl acetate extraction from the cartridge. High-performance liquid chromatography with amperometric detection was used for separation and detection of MHPG and the internal standard 3-ethoxy-4-hydroxyphenylglycol (EHPG). This method provided good, clean chromatograms with base-line separation of the appropriate peaks. This technique is sensitive, reliable and less time-consuming than other methods. Thus, only 0.5 ml of plasma is needed and the within-analysis and between-analysis coefficients of variation were 5.2% and 13% respectively. The plasma MHPG values in normal control were in good agreement with those using more complex methods.