HPLC-ELSD-UV法同时测定人体胆结石中胆固醇、胆红素及各种胆酸的含量

Study on Simultaneous Quantitative Determination of Cholesterol,Bilirubin and Various Cholic Acids in Human Gallstones by HPLC -ELSD -UV

目的:建立了人体胆结石中胆固醇、胆红素及各种胆酸的HPLC-ELSD-UV含量测定方法。方法:采用ZORBAX SB-C18(4.6mm×150 mm,5μg)色谱柱,0.1%醋酸溶液(A相)和甲醇(B相)为流动相梯度洗脱,流速:1.6 mL·min-1,紫外检测器(检测波长450 nm)和蒸发光散射检测器联用检测,柱温为室温。结果:测定了16个人体胆结石中胆固醇、胆红素及各种胆酸的含量。胆固醇在0.2204-1.653 mg·mL-1、胆红素在0.536-21.44μg·mL-1、各种胆酸在0.2-1.5 mg·mL-1范围内呈良好的线性关系;平均回收率分别为99.8%,101.5%,96.5%-101.2%,RSD分别为1.86%、1.71%、0.53%-2.14%(n=9);人体胆结石中的胆固醇、胆红素含量大多在776.3-972.5μg·mg-1和0.639-2.981μg·mg-1,各种胆酸未检出。结论:本法操作简便、快速、准确、适用范围广,可用于对人体胆结石中胆固醇、胆红素及各种胆酸的同时测定。

Objective:To establish an HPLC-ELSD - UV method to quantitative determine cholesterol,bilirubin and various cholic acids in human gallstones simultaneously. Method: The HPLC method was conducted on a ZOR BAX SB - C18 column(4. 6 mm×150 mm,5μm and coupled with ELSD and UV detector at 450 nm. The mobile phase was consisted of solvent A-0.1% acetic acid solution and solvent B-methanol with gradient elution program. The flow rate was 1. 6 mL·min-1 and the column temperature was room temperature. Result: Cholesterol, bilirubin and various cholic acids in human gallstones were quantitative determined simultaneously by the HPLC method. The linear range of cholesterol,bilirubin and various cholic acids were 0. 2204 - 1. 653 mg ·mL-1,0. 536 -21.44 μg·mL-1,0. 2 - 1. 5 mg·mL-1 respectively. The average recovery of cholesterol,bilirubin and various cholic acids were 99. 8% ,101. 5% ,96. 5% - 101. 2% respectively and their RSD were 1. 86% ,1.71% ,0. 53% -2. 14% ( n = 9 ) respectively. The assay results of cholesterol, bilirubin in human gallstones were in the range of 776. 3 -972. 5 μg·mg-1 and 0.639 -2.981μg·mg-1 ,and various cholic acids couldn' t be detected. Conclu tion:An accurate,fast,simple HPLC method to quantitative determine cholesterol,bilirubin and various cholic acids in human gallstones simultaneously was established.