SIMULTANEOUS LIQUID-CHROMATOGRAPHIC ANALYSIS FOR 4-HYDROXY-3-METHOXYMANDELIC ACID AND 4-HYDROXY-3-METHOXYPHENYLACETIC ACID IN URINE

  • 1 January 1980
    • journal article
    • research article
    • Vol. 26  (2) , 291-294
Abstract
A rapid, reliable high-performance liquid chromatographic method of simultaneously analyzing for 4-hydroxy-3-methoxymandelic acid (I) and 4-hydroxy-3-methoxyphenylacetic acid (II) in urine is presented. Paired-ion chromatography and amperometric detection are used. A 5-ml aliquot of [human] urine is adjusted to pH 6.1 and subjected to anion-exchange chromatography (Dowex AG 1-.times.4, 1.8 .times. 0.7 cm column). The 2 analytes are eluted from the column with 10 ml of 3 mol/l sodium chloride, and 2 .mu.l of the eluate, injected directly into the liquid chromatograph, is chromatographed at 40.degree. C. This column contains .mu.Bondapak C18, and the mobile phase is 100 mmol/l potassium phosphate buffer, pH 6.7, containing 0.5 mmol of tetrabutylammonium phosphate/l. Detector sensitivity is 50 nA full scale. Analytical recoveries of I and II are 80-84% and 48-49%, respectively; the coefficients of variation for various concentrations in urine are .apprx. 10%. [This procedure is applicable to biochemical diagnoses of neuroblastoma and pheochromocytoma.].