【摘要】 目的:建立测定人血清中硫酸吲哚酚浓度的高效液相色谱-荧光检测法,了解血液透析和血液透析滤过对硫酸吲哚酚的清除效果。方法:血清样品经乙腈沉淀蛋白;采用Hypersil BDS C18色谱柱(250mm×4.6mm,5μm);流动相为乙腈-水-三氟乙酸(15∶85∶0.2,V/V/V),流速为1mL/min;入射波长为280nm,发射波长为390nm。分别测定尿毒症患者透析前后血清中硫酸吲哚酚浓度。结果:线性范围为0.5~80.0μg/mL,线性方程为Y=2.438×104X+6.964×103,r=0.9986;日内、日间精密度均小于15%,方法回收率为92.8%~111.4%,提取回收率为90.9%。血液透析和血液透析滤过后患者血清中硫酸吲哚酚的下降率分别为(19.6±7.3)%和(25.3±7.0)%。结论:本法简便、快速、灵敏、准确,可用于测定人血清中硫酸吲哚酚浓度。更多还原

【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.更多还原