LIQUID-CHROMATOGRAPHIC ASSAY OF URINARY VANILLYLMANDELIC ACID AND HOMOVANILLIC-ACID, WITH CLEANUP BY ON-COLUMN INJECTION OF ACETONITRILE OR METHANOL

To separately measure vanillylmandelic acid (VMA) and homovanillic acid (HVA) in urine, the sample is diluted 10-fold with 50 mmol/L phosphoric acid, then exactly 5 .mu.L is injected directly onto a 50 .times. 4.6 mm column of Nucleosil 3C18. Samples can be injected at 5-min intervals because all peaks after these compounds of interest are washed away. VMA is eluted from the column after 2.8 min with 50 mmol/L phosphate buffer, pH 2.2, and measured by electrochemical detection. All peaks eluting after VMA are washed away together, by injection of 50 .mu.L of acetonitrile onto the column. HVA is eluted from the column after 3.3 min with a 100/10 (by vol) mixture of the phosphate buffer and acetonitrile. All later-eluting peaks are washed away together, by injection of 50 .mu.L of methanol onto the column. Analytical recoveries of VMA and HVA were 98.5% and 100.6%, respectively; the CVs for various concentrations of either in urine were about 3%.