Analysis of Human Urine by High Performance Liquid Chromatography with Ultraviolet Detection Using Octadecylsilica Column

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Ultraviolet absorbing compounds in human urine were analyzed by means of high-performance liquid chromatography using the column of octadecylsilica(TSK-gel ODS-120A, Toyo Soda). The sample was eluted with a linear gradient of 0.1 M potassium dihydrogen phosphate(pH, 2.12)to 0.1 M potassium dihydrogen phosphate-acetonitrile(1:1)at O-60 min and O.1 M potassium dihydrogen phosphate-acetonitrile(1:1)at 60-80 min and monitored with the absorption at 254 nm. Reproducible chromatograms were obtained and 55 well-resoluted peaks were numbered. A number of urine samples collected from healthy females with and without the administration of diuretics were analyzed, The correlation coefficients between every pair of peaks among the numbered peaks were calculated to examine the daily variations, the individual difference and the effects of diuretics on the UV-absorbing components. From the cross correlation matrix thus obtained, a few peaks which correlated closely with the peak of creatinine were identified. The results showed the feasibility of the HPLC analysis of urine concerning the diagnostic purposes.
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