The Assay of 4-Hydroxy-3-Methoxymandelic Acid in Urine by HPLC with Electrochemical Detection Using Bonded-Phase Silica Sorbents for Rapid, Simple and Selective Extraction

Abstract

A method is reported for the determination of 4-hydroxy-3-methoxymandelic acid (HMMA) by HPLC with electrochemical detection. The preparation of sample prior to HPLC has been studied and an efficient and selective extraction procedure described. Bonded-phase silica extraction columns and a vacuum manifold were used for the simple and rapid processing of batches of urine samples. Combining a reverse-phase C₁₈ and an anion exchange column ensures selective isolation of HMMA. This simplified greatly the subsequent chromatography. The method was combined into a simple scheme for the routine analysis of urine HMMA, catecholamines and 5-hydroxyindole-3-acetic acid. The HPLC was standardised such that a single mobile phase was used with minor modification for each of the assays.