In this study, a simple, fast and sensitive LC/MS/MS method was developed and validated for the determination of GW9508 in rat plasma. The sample was precipitated with acetonitrile and subsequently separated on ZORBAX Eclipse XDB C₁₈ column (50 mm × 2.1 mm, 5 μm). Mobile phase was composed of 0.1% formic acid in water and acetonitrile with gradient elution, at a flow rate of 0.4 mL/min. The analyte and internal standard were quantitatively monitored with precursor-to-product transitions of m/z 348.2→183.1 and m/z 397.2→260.2, respectively. The linearity of the assay was evident in the range of 1–1000 ng/mL with correlation coefficient more than 0.998. The validation parameters were all within the acceptable limits. The validated method has been successfully applied to the pharmacokinetics study of GW9508 in rat plasma, and our results demonstrated that GW9508 showed low clearance, moderate half-life and ideal bioavailability (54.88%). Furthermore, metabolites stemmed from rat plasma, rat hepatocytes and human hepatocytes were analyzed by an LC-Q-Exactive-Orbitrap-MS assay, resulting in the identification of seven metabolites based on the accurate mass and fragment ions. Acylglucuronide conjugate (M6) was found as the most abundant metabolite in all tested matrices. The metabolic pathways were proposed as hydroxylation and glucuronidation. This study provided an overview of disposition of GW9508, which is highly instructive for better understanding the effectiveness and toxicity of this drug.

在这项研究中，开发了一种简单，快速，灵敏的LC / MS / MS方法，并验证了其在大鼠血浆中测定GW9508的有效性。样品用乙腈沉淀，然后在ZORBAX Eclipse XDB C ₁₈色谱柱（50 mm×2.1 mm，5μm）上分离。流动相由水和乙腈中的0.1％甲酸组成，梯度洗脱，流速为0.4 mL / min。使用m / z 348.2→183.1和m / z的前体到产物的转变对分析物和内标进行定量监测分别从397.2→260.2。测定的线性在1–1000 ng / mL范围内明显，相关系数大于0.998。验证参数均在可接受的范围内。验证的方法已成功应用于大鼠血浆中GW9508的药代动力学研究，我们的结果表明GW9508具有低清除率，中等半衰期和理想的生物利用度（54.88％）。此外，通过LC-Q-Exactive-Orbitrap-MS分析法分析了源自大鼠血浆，大鼠肝细胞和人肝细胞的代谢物，从而基于准确的质量数和碎片离子鉴定出了七种代谢物。在所有测试的基质中，发现酰基葡萄糖醛酸苷共轭物（M6）是最丰富的代谢产物。代谢途径被提议为羟基化和葡萄糖醛酸化。