Homovanillic acid (HVA) was first identified in 1957 by Armstrong et al. (1) as a normal urinary constituent and a major metabolite of dopa and dopamine (2,3). Abnormal excretion of HVA in association with a variety of diseases including dopamine-secreting tumors (4-7)) familial dysautonomia (8), acute myocardial infarction (9), and certain nervous system disorders (10) has been described.
Although a few investigators have achieved success with high-voltage electrophoresis (11) and thin-layer chromatography ( 12)) most t,echniques for the measurement of HVA have involved paper chromatography (2,13-15)) calorimetry ( 16)) spectrophotometry ( 17)) fluorometry (8,1%20), and a combination of paper chromatography and fluorometry (21). Early attempts to apply gas-liquid chromatography to the detection of HVA were insensitive or time-consuming (22,23). The experience of this laboratory with gas-liquid chromatography and electron capture detection (24,25) led to the development of a procedure in which subnanogram amounts of an hexafluoroisopropyl (HFIP) derivative of trifluoroacetylated homovanillic acid (HVA-TFA) can be readily and reliably measured in human urine. HVA excretions by normal subjects are presented and compared. METHODS 1 ml of urine was placed in a 15 ml ground-glass stoppered centrifuge tube, acidified with 0.3 ml 6 iv hydrochloric acid (pH < 1)) diluted to 3 ml with distilled water, saturated with sodium chloride (1 gm), and 1 Part of this work is taken from the thesis of Stanley W. Dziedzic submitted to the Graduate School of Fordham University in partial fulfillment of the requirements for the Ph.D. degree in chemistry, 1972.