反相高效液相色谱法测定结合胆汁酸及肝硬化患者血清结合型胆汁酸分析

Determining Conjugated Bile Acids by Reverse HPLC and Analyzing the Conjugated Bile Acids in Sera of Patients with Hepatocirrhosis.

目的 通过反相高效液相色谱法 (HPL C)测定血清中结合型胆汁酸的亚组分并探讨肝硬化患者结合型胆汁酸亚组分的变化。方法 用反相 HPL C测定结合型胆汁酸的亚组分 ,并评价该方法的精密度、准确度、灵敏度及线性范围。测定肝硬化患者血清中的 6种结合型胆汁酸并与对照组进行比较。结果 反相 HPL C法测定胆汁酸各组分的最低检测限为 3.12 5μm ol/ L ;在浓度为 3.5～ 10 0μmol/ L内 ,其日内变异小于 10 % ,日间变异小于15 % (除 TL CA外 ) ,各胆汁酸的回收率在 93.3%～ 10 6 .1%之间 (除牛磺石胆酸外 )。肝硬化患者血清中的胆汁酸水平明显高于正常对照 (P<0 .0 5 )。结论 反相 HPL C法适用于临床血清胆汁酸的分析。肝硬化患者血中胆汁酸浓度可为临床诊断和治疗提供帮助。

Objective To investigate the conjugated bile acids in sera of patients with hepatocirrhosis by the determination of conjugated bile acids using reverse HPLC(high performance liquid chromatography). Methods Reverse HPLC was used to determine the conjugated bile acids, to evaluate the precision, accuracy, sensitivity and linear limitation of the method, and to separate and determine six conjugated bile acids in the sera of hepatocirrhotic patients. A comparision with controls was performed. Results The limitation of determination was 3.125 μmol/L. The coefficients of variation were less than 10% within day and less than 15% day to day for the concentration (3.5 100 μmol/L) of bile acids, and the recoveries were 93.3% 106.1% except TLCA. The concentrations of conjugated bile acids in sera of patients with hepatocirrhosis were significantly higher than those of the control group (P<0.05). Conclusion The method can meet the need of the determination of conjugated bile acids in serum. The concentrations of conjugated bile acids in sera of patients with hepatocirrhosis are of use for clinical diagnosis and treatment.