A rapid and sensitive UPLC-MS/MS method was developed and fully validated for the quantification of hyperoside in rat plasma after intragastric, intraperitoneal and intravenous administration. Geniposide was used as an internal standard, and simple liquid–liquid extraction by ethyl acetate was utilized for to extracting the analytes from the rat plasma samples. Chromatographic separation was carried out on an InfinityLab Poroshell 120EC-C18column (2.1 mm × 50 mm, 1.9-Micro, Agilent technologies, USA). The mobile phase consisted of methanol (A) and water (B) (containing 0.1% acetic acid) at a flow rate of 0.4 mL/min. A run time of 3 min for each sample made it possible to analyze more than 300 plasma samples per day. The validated linear ranges of hyperoside were 2–1000 ng/mL in rat plasma. The intra-day and inter-day precision were within 2.6–9.3%, and accuracy were ± 8.6%. And the results of recovery and matrix interference studies were well within the accepted variability limits. Finally, this method was fully validated and successfully applied to the pharmacokinetic studies of hyperoside via different administration routes in rats.

中文翻译：

---

UPLC-MS/MS法测定金丝桃苷及其在大鼠不同给药途径后药代动力学研究中的应用

开发并充分验证了一种快速灵敏的 UPLC-MS/MS 方法，用于定量大鼠灌胃、腹膜内和静脉内给药后血浆中金丝桃苷的含量。以栀子苷为内标，利用乙酸乙酯进行简单的液-液萃取，从大鼠血浆样品中提取分析物。在 InfinityLab Poroshell 120EC-C18 色谱柱（2.1 mm × 50 mm，1.9-Micro，Agilent Technologies，USA）上进行色谱分离。流动相由甲醇 (A) 和水 (B)（含有 0.1% 乙酸）组成，流速为 0.4 mL/min。每个样品的运行时间为 3 分钟，因此每天可以分析 300 多个血浆样品。大鼠血浆中金丝桃苷的验证线性范围为 2-1000 ng/mL。日内和日间精度在2.6-9.3%之间，准确度为±8.6%。回收率和基质干扰研究的结果完全在可接受的变异范围内。最后，该方法通过不同给药途径在大鼠体内得到充分验证并成功应用于金丝桃苷的药代动力学研究。