【摘要】 目的:建立测定人胆汁中结合型胆汁酸的HPLC-紫外检测法。方法:胆汁样品经固相萃取,采用WatersNovaC18色谱柱,柱温为25℃,流动相为甲醇∶磷酸盐缓冲液(60mmol/L,pH3.3)(66∶34,v/v),流速0.8ml/min,检测波长197nm。结果:9种结合型胆汁酸在20min内分离完全,各胆汁酸在浓度为8~1000μmol/L范围内线性良好,r为0.9991~0.9999;最低检测限为1.25~3.13μmol/L;该方法日内变异在3.77%~8.20%之间,日间变异在6.73%~17.11%之间,各胆汁酸的回收率在88.23%~109.75%之间。结论:本方法准确、灵敏、快速,适用于胆汁中结合型胆汁酸水平的实验室研究和临床常规检测。更多还原

【Abstract】 Objective:To develop a HPLC method with ultraviolet detection to determine nine types of conjugated bile acids in human bile.Methods:Bile acids in bile juice were extracted using Sep-Pak C18 cartridges.The analytes were eluted using methanol/phosphate buffer(60 μmol/L,pH3.3)(66:34,v/v)as mobile phase on a Waters Nova C18 column.The flow rate was 0.8 ml/min.The column was kept at 25℃.The eluate was monitored by a ultraviolet detection at 197nm.Results:The nine types of conjugated bile acids were completely analyzed in twenty minitus by this method.The assay was linear in the range 8~100 0 μmol/L,with the correlation coeffients for linear regression from 0.9991 to 0.999 9.The detection limits ranged from 1.25 to 3.12 μmol/L.The within-day precision was below 8.20%,and the between-day precision was below 17.11%.The recoveries were 88.23%~109.75%.Conclusion:This method is accurate,sensitive,rapid and suitable for the rountine clinical practice and research practice.更多还原