Division of Pharmaceutics and Pharmacology, College of Pharmacy, The Ohio State University, Columbus, OH, USA.
Clinical Pharmacology Program, Office of the Clinical Director, National Cancer Institute, Bethesda, MD, USA.
J Chromatogr B Analyt Technol Biomed Life Sci. 2022 Nov 1;1210:123437. doi: 10.1016/j.jchromb.2022.123437. Epub 2022 Aug 27.
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 (r > 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.
甘氨胆酸-3-硫酸盐 (GCDCA-S) 和鹅去氧胆酸-24-葡萄糖醛酸 (CDCA-24G) 是可能作为肝摄取转运蛋白 OATP1B1 和 OATP1B3 介导的药物相互作用的内源性生物标志物的胆汁酸代谢物。我们开发并验证了一种用于定量测定小鼠和人血浆中 GCDCA-S 和 CDCA-24G 的新型 UHPLC-MS/MS 方法，定量下限为 0.5 ng/mL。色谱分离在 Accucore aQ 柱（50mm×2.1mm，dp=2.6μm）上进行，柱温保持在 20°C，梯度流动相由 2mM 氨乙酸在水中和甲醇组成。GCDCA-S 和 CDCA-24G 的提取回收率均>80%，线性（r>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）。分析物加标质控样品的精密度（CV<11.6%）和准确度偏差（10.9%）值证明水是制备校准品的可接受基质。该方法成功应用于建立人类和小鼠 OATP1B1/OATP1B3 的基线活性，并建立 OATP1B1/OATP1B3 抑制剂利福平米卡芬净的体内作用。
