Quntitative determination of oxypurines in biological samples by high-performance liquid chromatography

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A method for the quantitative determination of oxypurines in biological samples has been developed by reversed - phase high - performance liquid chromatography. Eluate from a column was amperometrically and spectrophotometrically monitored by aid of an electrochemical detector and an ultraviolet absorption detector, respectively. The electrochemical detection method has been found to be highly selective and sensitive compared with the ultraviolet absorption detection method. However, the amperometric detection method allows to determine not hypoxanthine, but xanthine and uric acid in biological samples because hypoxanthine cannot be effectively oxidized under the present chromatographic conditions. On the other hand, the spectrophotometric detection method allows to determine hypoxanthine, xanthine and uric acid in biological samples by one time chromatography showing its low detectability and selectivity. In addition, we have found that allopurinol cannot be oxidized under the present chromatographic conditions, whereas its metabolite (oxipurinol) can be oxidized. We have emphasized that those chromatographic methods developed in this study are the powerful tools for a better understanding of disorder (s) of purine nucleotides metabolism in relation with some diseases such as cancer and tissue infarction. Effectiveness of the liquid chromatographic methods has been also demonstrated showing some reversed - phase liquid chromatographic results with some other related results already published by us. For examples, the liquid chromatographic examinations have revealed that urate level is lower in the hepatomous tissue than its surrounding normal area, whereas xanthine level is higher in the former than in the latter. Our finding is consistent with the earlier finding by Prajda et al. that xanthine oxidase was decreased in all hepatomas. The electrochemical detection method allows to determine not only oxypurines but also any other electroactive biological compounds such as glutathione, tyrosine and methionine.
Japanese Society of Gout and Nucleic Acid Metabolismの論文

Quntitative determination of oxypurines in biological samples by high-performance liquid chromatography

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