Clinical Pharmacology Research Laboratory, The First Affiliated Hospital of Soochow University, Suzhou, China.
Department of Pharmaceutics, College of Pharmaceutical Science, Soochow University, Suzhou, China.
J Pharm Biomed Anal. 2018 Feb 20;150:287-293. doi: 10.1016/j.jpba.2017.12.010. Epub 2017 Dec 13.
A rapid and highly sensitive liquid chromatography-tandem mass spectrometry (LC-MS/MS) method has been developed and validated for the determination of a novel anti-HBV compound Y101 and its metabolites M8 and M9 in human plasma. The plasma samples were deproteinated with acetonitrile after addition of Peramivir (internal standard, IS) and separated on a 40 °C ACQUITY UPLC BEH C column (2.1 mm × 50 mm, 1.7 μm). The mobile phase consisted of water (containing 5 mM ammonium acetate and 0.1% formic acid) and acetonitrile (74:26, v/v) at a flow rate of 0.3 mL/min. The detection was performed on a Triple Quad 5500 tandem mass spectrometer coupled with electrospray ionization (ESI) source in positive mode. Quantification was conducted by multiple reaction monitoring (MRM) of the transitions of m/z 490.1 → 339.0 for Y101, m/z 357.2 → 105.2 for metabolite M8, m/z 373.1 → 105.1 for metabolite M9 and m/z 329.1 → 270.2 for IS, respectively. The method was validated over the calibration curve range of 1.000-1000 ng/mL for Y101, 2.000-2000 ng/mL for metabolite M8 and 0.3000-300.0 ng/mL for metabolite M9, using linear regression and 1/x weighting. No matrix effect and carryover effect was observed. The intra- and inter-batch precision and accuracy of Y101, metabolite M8 and M9 were all within the acceptable criteria. This method allows a rapid and simple determination of Y101 and its metabolites M8 and M9 in human plasma. It was successfully applied in a pharmacokinetic study in human for the first time.
建立并验证了一种用于测定人血浆中新型抗乙肝化合物 Y101 及其代谢物 M8 和 M9 的快速、高灵敏度的液相色谱-串联质谱（LC-MS/MS）方法。在加入金刚烷胺（内标，IS）后，用乙腈对血浆样品进行蛋白沉淀处理，在 40°C ACQUITY UPLC BEH C 柱（2.1mm×50mm，1.7μm）上进行分离。流动相由水（含 5mM 乙酸铵和 0.1%甲酸）和乙腈（74:26，v/v）组成，流速为 0.3mL/min。检测采用电喷雾电离（ESI）源正模式下的三重四极杆 5500 串联质谱仪进行。通过 m/z 490.1→339.0 的 Y101、m/z 357.2→105.2 的代谢物 M8、m/z 373.1→105.1 的代谢物 M9 和 m/z 329.1→270.2 的 IS 的多重反应监测（MRM）进行定量。该方法在 Y101 的校准曲线范围为 1.000-1000ng/mL、代谢物 M8 为 2.000-2000ng/mL、代谢物 M9 为 0.3000-300.0ng/mL 时进行了验证，采用线性回归和 1/x 加权。未观察到基质效应和交叉污染效应。Y101、代谢物 M8 和 M9 的批内和批间精密度和准确度均在可接受范围内。该方法可快速简便地测定人血浆中的 Y101 及其代谢物 M8 和 M9。该方法首次成功应用于人体药代动力学研究。
