Liquid chromatography with multichannel ultraviolet detection used for studying disorders of purine metabolism

We used a reversed-phase "high-performance" liquid-chromatographic system equipped with a multichannel ultraviolet spectrometric detector and a micro-computer for analyzing urine samples from patients with disorders of purine metabolism. This system recorded a series of absorption-spectrum data from a single chromatographic run and stored them for subsequent analysis. Because the retention times and ultraviolet absorption spectra of the eluates were recorded simultaneously, identification of peaks was easy and quite accurate for simultaneous quantification of orotidine, adenine, hypoxanthine, uric acid, xanthine, allopurinol (4-hydroxypyrazolo[3,4-d]pyrimidine), oxypurinol (4,6-dihydroxypyrazolo[3,4-d]pyrimidine), inosine, and 2,8-dihydroxyadenine--compounds extremely difficult or even impossible to quantify simultaneously with a conventional single-wavelength spectrometer. We used this method to investigate purine metabolites in urines from a patient with hereditary xanthinuria, three patients with 2,8-dihydroxyadenine urolithiasis, and a gouty subject taking allopurinol.