Abstract
A quantitative chemical method for the determination of urinary d-glucaric acid is described. Following the separation of d-glucaric acid by ion-exchange column chromatography using Dowex 1 X-8 borate, a portion of the sample is treated under neutral conditions (pH ca. 6.8) with periodic acid; the glyoxylic acid obtained is condensed with phenylhydrazine. The glyoxylic acid phenylhydrazone thus formed undergoes mild oxidation with potassium ferricyanide in the presence of excess phenylhydrazine, resulting in the intensely colored 1,5-diphenylformazan, the absorbancy of which is determined at 520 mμ. This method has been applied to determine d-glucaric acid excretion in human, rat, and guinea-pig urine; the results are compared with those obtained by the enzymic method of Marsh.
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