Determination of indoxyl sulfate in human serum by HPLC-FLU and Its application in hemodialysis patients

Abstract

Abstract: AIM: To establish a high-performance liquid chromatography method with fluorescence detector for the determination of indoxyl sulfate (IS) in human serum, and apply to study the clearance effect of IS by hemodialysis and hemodiafiltration.METHODS: The serum was deproteinated by acetonitrile. The chromatographic separation was achieved on a Hypersil BDS C₁₈ column (250 mm×4.6 mm, 5 μm) with the mobile phase consisted of acetonitrile-water-trifluoroacetic (15∶85∶0.2, V/V/V) at a flow rate of 1 mL/min. The fluorescence detector was operated at an excitation and emission wavelengths 280 nm and 390 nm, respectively. The serum level of IS was deteated before and after dialysis in uremic patients.RESULTS: A good linearity was obtained from 0.5 to 80.0 μg/mL with r=0.9986, the linear regression equation was Y=2.438×10⁴X+6.964×10³. The intra-day RSD and inter-day RSD were both less than 15%. The method recoveries were 92.8%-110.4% and the extraction recovery was 90.9%. The reduction ratios of IS after hemodialysis and hemodiafiltration were (19.6±7.3)% and (25.3±7.0)%, respectively. CONCLUSION: The method is simple, sensitive and accurate for the determination of indoxyl sulfate in serum.

Key words: Indoxyl sulfate, HPLC, Fluorescence detection, Hemodialysis

CLC Number:

R969. 1

WU Chun, SHI Cheng-gang, LI Mei, PAN Bi-yan, XING Xiao-min, FENG Zhi-qiang, XIE Zhi-yong. Determination of indoxyl sulfate in human serum by HPLC-FLU and Its application in hemodialysis patients[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2010, 15(8): 906-910.

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