Ye Lian, Liu Suya, Wang Meng, Shao Yong, Ding Min
Key Laboratory of Clinical Laboratory Diagnostics, Ministry of Education, Chongqing Medical University, Chongqing 400016, China.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Dec 1;860(1):10-7. doi: 10.1016/j.jchromb.2007.09.031. Epub 2007 Oct 2.
A simple, sensitive, and specific high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS) method for the analysis of the bile acid profile has been developed. Fifteen bile acids, including free and conjugated bile acids, were separated and detected by HPLC-MS/MS. The MS detection was performed by electrospray ionization (ESI) in negative ion mode. Quantification was achieved in multiple reaction monitoring (MRM) mode with external standard curve methods. Total analysis time was 15 min for one sample including re-equilibration time of the column. The assay was linear in the range 0.02-100.0 micromol/L with correlation coefficients of standard curves for all bile acids better than 0.999. The detection limits ranged from 0.001 to 0.006 micromol/L for different bile acids. The precisions for each bile acid were CVs<3.8% for within-day and CVs<6.1% for between-day. The average recoveries for all bile acids studied were in the range of 86-110.0%. The developed method was applied to the analysis of clinic samples consisting of 53 women with healthy pregnancies and 43 women with intrahepatic cholestasis of pregnancy (ICP). The results revealed that the bile acid profile was markedly different between women with ICP and women with healthy pregnancies.
已开发出一种用于分析胆汁酸谱的简单、灵敏且特异的高效液相色谱-串联质谱法（HPLC-MS/MS）。通过HPLC-MS/MS分离并检测了15种胆汁酸，包括游离胆汁酸和结合胆汁酸。质谱检测采用电喷雾电离（ESI）负离子模式进行。采用外标曲线法在多反应监测（MRM）模式下进行定量分析。包括色谱柱重新平衡时间在内，一个样品的总分析时间为15分钟。该测定法在0.02 - 100.0 μmol/L范围内呈线性，所有胆汁酸标准曲线的相关系数均优于0.999。不同胆汁酸的检测限在0.001至0.006 μmol/L之间。每种胆汁酸的日内精密度变异系数（CV）<3.8%，日间精密度变异系数（CV）<6.1%。所研究的所有胆汁酸的平均回收率在86 - 110.0%范围内。所开发的方法应用于对53例正常妊娠女性和43例妊娠肝内胆汁淤积症（ICP）女性的临床样本分析。结果显示，ICP女性和正常妊娠女性的胆汁酸谱存在显著差异。
