Saponin PH, akemisaponins E, saponin PJ1 and scheffoleoside A, the main bioactive triterpene saponins of Chinese traditional medicine Akebia trifoliata, contribute to its diuretic pharmacological activity. Because of interactions of the multiple ingredients in vivo, pharmacokinetic studies of multiple triterpenes after administration of A. trifoliata extract are essential to clarify their pharmacological effects. The purpose of this study was to develop an efficient and sensitive UHPLC-MS/MS method for simultaneous determination of these four triterpene saponins in rat plasma. The biosamples were prepared by liquid-liquid extraction with n-butanol. The chromatographic separation was performed on a Phenomenex Luna® C18 (150 × 2 mm, 3 μm) with a mobile phase consisting of acetonitrile and water at a flow rate of 0.5 mL/min. The MS/MS system was operated in a negative multiple reaction monitoring mode, and the precursor-product ion transitions were optimized as m/z 941.6 → 471.1 for saponin PH, 941.7 → 471.2 for akemisaponins E, 1089.7 → 601.1 for saponin PJ1 , 957.6 → 487.4 for scheffoleoside A and 799.5 → 637.3 for ginsenoside Rg1 (Rg1 , internal standard). Method validation parameters (calibration curve linearity, lower limit of detection, recovery, matrix effect, intra- and inter-day precision) were within the acceptable ranges. This is the first reported on the UHPLC-MS/MS detection of saponin PH, akemisaponins E, saponin PJ1 and scheffoleoside A, and applied to a preclinical pharmacokinetic study after oral administration of A. trifoliata extract in rats. This study provides a basis for clinical application and further development of A. trifoliata extract.

中文翻译：

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经验证的UHPLC-MS / MS方法可同时测定大鼠血浆中三叶木通提取物中的四种三萜皂苷及其在药代动力学研究中的应用。

皂苷PH，akemisaponins E，皂苷PJ1和鹅掌苷A是中草药三叶草的主要生物活性三萜皂苷，有助于利尿药理活性。由于多种成分在体内的相互作用，三叶草提取物给药后多种三萜的药代动力学研究对于阐明其药理作用至关重要。这项研究的目的是开发一种高效，灵敏的UHPLC-MS / MS方法，用于同时测定大鼠血浆中的这四种三萜皂苷。通过用正丁醇液-液萃取制备生物样品。色谱分离在Phenomenex C18（150×2 mm，3μm）上进行，流动相由乙腈和水组成，流速为0.5 mL / min。MS / MS系统在负多反应监测模式下运行，并且对前体产物离子的跃迁进行了优化，皂苷PH的m / z为941.6→471.1，那米皂苷E为941.7→471.2，皂苷PJ1为1089.7→601.1，957.6 →鹅掌ole苷A为487.4，人参皂苷Rg1为799.5→637.3（内标Rg1）。方法验证参数（校准曲线线性，检测下限，回收率，基质效应，日内和日间精度）在可接受的范围内。这是首次报道用UHPLC-MS / MS检测皂苷PH，akemisaponins E，皂苷PJ1和鹅掌苷A，并在大鼠口服三叶草提取物后应用于临床前药代动力学研究。该研究为三叶草提取物的临床应用和进一步开发提供了依据。