Mass fragmentographic assay of homovanillic acid in brain tissue

Abstract— Available methods for the determination of homovanillic acid (HVA) are based on fluorimetric measurements. The present method uses gas chromatography-mass spectrometry with the aid of deuterium labelled HVA methyl ester as a carrier and internal standard. Quantitation is achieved by comparing intensities of the molecular ion (= base peak) of the protium (H) and deuterium (D) forms of the methyl ester heptafluorobutyric derivatives of HVA. A standard curve is constructed by measuring mixtures of known amounts of protium and deuterium derivatives and plotting peak height ratios H/D on the ordinate and ratios of amounts H/D on the abscissa. Analysis of ten individual mouse brains gave a mean value of 1·36 nmole/g brain tissue with a standard deviation of ±9 per cent. Chlorpromazine elevates, whereas pargyline reduces the level of HVA in brain markedly. The described procedure offers advantages because of the greatly increased specificity and sensitivity permitting analyses in discrete brain areas.