Evaluation and improvement of homovanillic acid methodology, and a critique of liquid chromatography with electrochemical detection.
Resolution of several problems encountered with some organic solvent extraction-fluorometric procedures commonly used to measure homovanillic acid (HVA) in urine resulted in a more specific and sensitive methodology, applicable to diverse biological material. Purification on an anion-exchange resin followed by extraction with organic solvent was the most reliable and specific for HVA. Eluting the resin with a near-boiling solution of NaCl concentrated the HVA in a smaller volume with no increase in the blank reading. In none of the other methods was enough oxidizing agent used to completely oxidize the excessive amounts of HVA in urines of patients with neuroblastoma. 5-Hydroxyindoleacetic acid interferes with the HVA fluorophor. Several of the advantages and problems with "high-pressure" liquid chromatography with electrochemical detection are identified, particularly the unpredictable nature of the working electrode.
D002853:Chromatography, Liquid / Q000379:methods; D004563:Electrochemistry; D005456:Fluorescent Dyes; D005470:Fluorometry; D006719:Homovanillic Acid / Q000652:urine*; D006801:Humans; D006897:Hydroxyindoleacetic Acid / Q000652:urine
D005456:Fluorescent Dyes; D006897:Hydroxyindoleacetic Acid; D006719:Homovanillic Acid