A sensitive and robust LC-MS/MS method has been developed and validated to simultaneous determine the concentrations of tumulosic acid, dehydrotumulosic acid, polyporenic acid C, cinnamic acid, Atractylenolide I, Atractylenolide II, Atractylenolide III, glycyrrhizic acid, glycyrrhetinic acid, liquiritigenin and isoliquiritin in plasma from rats who received Ling-Gui-Zhu-Gan Decoction extract oral administration. The samples were prepared by a liquid-liquid extraction procedure using ethyl ether as the extraction solvent and schisandrin as the internal standard. Chromatographic separation was achieved using a Thermo Hypersil GOLD C₁₈ column (2.1 mm × 100 mm, 1.9 µm) and a gradient mobile phase consisting of acetonitrile-water with 0.1% formic acid. All of the analytes were quantified using negative and positive multiple reaction monitoring mode. The method was validated for selectivity, linearity, accuracy, precision, recovery, matrix effect and sample stability under various storage conditions, whose values are all fell in the acceptable limits. We report the lowest limit of quantification for tumulosic acid, dehydrotumulosic acid and polyporenic acid C as 2 ng/mL. This is the first study for simultaneous determination of so many analytes in rat plasma after oral administration of Ling-Gui-Zhu-Gan Decoction. This validated method was successfully used to study the pharmacokinetics of multiple compounds in rat plasma after oral administration of Ling-Gui-Zhu-Gan Decoction.

已开发并验证了灵敏且稳健的LC-MS / MS方法，可同时测定亚mul酸，脱氢肿瘤酸，多孔酸C，肉桂酸，白术内酯I，白术内酯II，白术内酯III，甘草酸，甘草次酸，脂白蛋白原灵桂竹甘汤口服液大鼠血浆中的异黄酮和异黄酮含量。通过液-液萃取程序，以乙醚为萃取溶剂，以五味子苷为内标，制备样品。使用Thermo Hypersil GOLD C ₁₈进行色谱分离色谱柱（2.1 mm×100 mm，1.9 µm）和由乙腈-水与0.1％甲酸组成的梯度流动相。使用阴性和阳性多重反应监测模式对所有分析物进行定量。验证了该方法在各种存储条件下的选择性，线性，准确性，精密度，回收率，基质效应和样品稳定性，其值均在可接受的范围内。我们报告了最低分子量为2 ng / mL的最小分子量为tumulosic acid，dehydrotumulicicic acid和polyporenic acidC。这是首次在口服灵桂竹肝汤后同时测定大鼠血浆中多种分析物的研究。该经验证的方法已成功用于研究口服口服灵桂竹肝汤后多种化合物在大鼠血浆中的药代动力学。