Abstract
Uric acid is generated by the oxidation of purines, but no enzyme is present to oxidize it further. It is present in plasma at a concentration approaching 500 J.lIIlollL and has been proposed as an important antioxidant. 1 In agreement with this proposal experiments in vitro have shown the protective effect of uric acid against oxidative damage. The first and major product of uric acid oxidation is allantoin, and it has been suggested that allantoin may be useful as a 'marker' of free radical reactions in vivo.2,3 However, methods currently used for the analysis of allantoin in body fluids are tedious and complicated, and since allantoin is very polar' it is difficult to obtain resolution from other serum constituents that have a similar polarity on a reverse-phase HPLC column. In the method of Grootveld and Halliwelluric acid is first analysed by HPLC, and allantoin is then measured in a separate procedure which involves initial separation and fraction collection by HPLC, hydrolysis and derivatization of allantoin, and finally separation of the derivatives by HPLC. We have developed a unique chromatographic assay to determine uric acid and allantoin simultaneously in plasma ultrafiltrate which is a significant improvement on previous HPLC methods.
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