Liquid chromatography at 3-methoxy-4-hydroxyphenylethylene glycol in urine with fluorescence detection

We describe a liquid-chromatographic procedure for determining low concentrations of 3-methoxy-4-hydroxyphenylethylene glycol in urine. After liquid-liquid extraction and cleanup of the sample, an aliquot of the extract is injected onto a reversed-phase chromatographic column, with isocratic elution with an acetonitrile/acetate buffer mobile phase. An on-line fluorescence detector is used for measuring the natural fluorescence of the compound. The method gives a relatively clean extract, which is suitable for measuring the less-than-normal concentrations associated with mentally depressed patients. Although a liquid-chromatographic technique has been recently described for measuring this compound in urine, the method involves electrochemical detection, which may not be available in many clinical laboratories, and, because of its nonspecificity, the method is only suitable for detecting the increased amounts of 3-methoxy-4-hydroxyphenylethylene glycol excreted by patients with pheochromocytoma.