Qi Dongli, Yang Xiaolin, Chen Jing, Li Fei, Shi Xiupu, Zhang Chunfeng, Yang Zhonglin
Key Laboratory of Modern Chinese Medicines, China Pharmaceutical University, Ministry of Education, No. 24 Tongjiaxiang, Nanjing, 210009, China.
Biomed Chromatogr. 2013 Nov;27(11):1568-73. doi: 10.1002/bmc.2961. Epub 2013 Jul 5.
A sensitive liquid chromatography-electrospray ionization-mass spectrometry method has been developed and validated for determination of two major bioactive saponins in rat plasma after oral administration of saponins extracted from Rhizoma Panacis Japonici, including chikusetsusaponin V and chikusetsusaponin IV for the first time. Akebia saponin D was used as the internal standard (IS). Plasma samples were prepared by protein precipitation with methanol. A Phenomenex C18 column (150 × 4.6 mm, 4 µm) was used as the analytical column with a mobile phase of acetonitrile and 0.05% aqueous formic acid. Mass spectrometric detection was achieved by single quadrupole mass spectrometer equipped with an electrospray ionization interface operating in negative ionization mode. Calibration curves showed good linearity over the concentration range of 5-500 ng/mL for the two analytes in rat plasma. The lower limit of quantification was 5 ng/mL. The intra- and inter-batch precisions were within 10.3% and accuracy ranged from -3.9 to 5.4%. The method was validated and successfully applied to the preliminary pharmacokinetic study of chikusetsusaponin V and chikusetsusaponin IV in rat plasma after oral administration of saponins extracted from Rhizoma Panacis Japonici.
已开发并验证了一种灵敏的液相色谱 - 电喷雾电离 - 质谱法，用于首次测定大鼠口服从竹节参中提取的皂苷后血浆中的两种主要生物活性皂苷，包括竹节人参皂苷V和竹节人参皂苷IV。以木通皂苷D作为内标（IS）。血浆样品通过用甲醇进行蛋白沉淀来制备。使用Phenomenex C18柱（150×4.6 mm，4 µm）作为分析柱，流动相为乙腈和0.05%甲酸水溶液。通过配备电喷雾电离接口并在负离子模式下运行的单四极杆质谱仪进行质谱检测。校准曲线表明，大鼠血浆中两种分析物在5 - 500 ng/mL浓度范围内具有良好的线性。定量下限为5 ng/mL。批内和批间精密度均在10.3%以内，准确度范围为 - 3.9%至5.4%。该方法经过验证，并成功应用于大鼠口服从竹节参中提取的皂苷后血浆中竹节人参皂苷V和竹节人参皂苷IV的初步药代动力学研究。
