PDF(Pubmed)
Abstract：
Glycochenodeoxycholate-3-sulfate (GCDCA-S) and chenodeoxycholate-24-glucuronide (CDCA-24G) are bile acid metabolites that potentially serve as endogenous biomarkers for drug-drug interactions mediated by the hepatic uptake transporters OATP1B1 and OATP1B3. We developed and validated a novel UHPLC-MS/MS method for the quantitative determination of GCDCA-S and CDCA-24G in mouse and human plasma with a lower limit of quantitation of 0.5 ng/mL. Chromatographic separation was achieved on an Accucore aQ column (50 mm × 2.1 mm, dp = 2.6 μm) maintained at 20 °C and a gradient mobile phase comprising 2 mM ammonium acetate in water and methanol. The extraction recoveries of GCDCA-S and CDCA-24G were >80 %, and linear (r2 > 0.99) calibration curves ranged 0.5-100 ng/mL (CDCA-24G and GCDCA-S in mouse plasma) or 0.5-1000 ng/mL (GCDCA-S in mouse plasma). Values for precision (CV < 11.6 %) and accuracy bias (10.9 %) of analyte-spiked quality control samples verified that water was an acceptable matrix to prepare calibrators. This method was successfully applied to establish baseline activity of OATP1B1/OATP1B3 in humans and mice and establish the in vivo effects of OATP1B1/OATP1B3 inhibitors rifampin and micafungin.
摘要：
硫酸甘氨酸脱氧胆酸盐-3-硫酸盐（GCDCA-S）和鹅脱氧胆酸盐-24-葡糖苷酸（CDCA-24G）是胆汁酸代谢物，可能作为内源性生物标志物，用于由肝脏摄取转运蛋白OATP1B1和OATP1B3介导的药物-药物相互作用。我们开发并验证了一种新型的UHPLC-MS/MS方法，用于定量测定小鼠和人血浆中的GCDCA-S和CDCA-24G，定量下限为0.5ng/mL。在AccucoreaQ色谱柱上实现色谱分离（50mm×2.1mm，dp=2.6μm）保持在20°C和包含2mM乙酸铵在水和甲醇中的梯度流动相。GCDCA-S和CDCA-24G的提取回收率均>80%,和线性（r2>0.99）校准曲线范围为0.5-100ng/mL（小鼠血浆中的CDCA-24G和GCDCA-S）或0.5-1000ng/mL（小鼠血浆中的GCDCA-S）。掺入分析物的质量对照样品的精密度(CV<11.6%)和准确度偏差(10.9%)的值证实水是制备校准物的可接受的基质。该方法已成功应用于建立OATP1B1/OATP1B3在人和小鼠中的基线活性，并建立了OATP1B1/OATP1B3抑制剂利福平和米卡芬净的体内作用。
