Simultaneous determination of 35 constituents and elucidation of effective constituents in a multi-herb Chinese medicine formula Xiaoer-Feire-Kechuan

Xiaoer-Feire-Kechuan(XFK) is an 11-herb Chinese medicine formula to treat cough and pulmonary inflammation.The complicated composition rendered its chemical analysis and effective-component elucidation.In this study,we combined quantitative analysis and bioactivity test to reveal the antiinflammatory constituents of XFK.First,UPLC-DAD and UHPLC/Q-Orbitrap-MS methods were established and validated to quantify 35 analytes(covering 9 out of 11 herbs) in different XFK formulations.Parallel reaction monitoring mode built in Q-Orbitrap-MS was used to improve the sensitivity and selectivity.Then,anti-inflammatory activities of the 35 analytes were analyzed using in vitro COX-2 inhibition assay.Finally,major analytes forsythosides H,I,A(8-10),and baicalin(15)(total contents varied from 21.79 to 91.20 mg/dose in different formulations) with significant activities(inhibitory rate ≥ 80%) were proposed as the anti-inflammatory constituents of XFK.The present study provided an effective strategy to discover effective constituents of multi-herb formulas.

正向×反相离线二维液相色谱-轨道质谱联用检测通脉颗粒中的微量成分

通脉颗粒是由丹参、川芎、葛根三味中药的水提物制成的用于治疗心血管疾病的复方制剂,主要成分为异黄酮苷、异黄酮及酚酸类。低极性化合物如丹参酮和酚酞类也可能对活性有贡献,但很难检测到。因此,本研究建立了正向×反相离线二维液相色谱方法对通脉颗粒中的有效成分进行富集和分离。采用正向SEP-PAK硅胶固相萃取柱分离通脉颗粒的乙酸乙酯萃取物,分别以石油醚、乙酸乙酯–甲醇(6:1,v/v)、甲醇洗脱获得三个流分,然后采用超高效液相反向色谱串联四级杆-静电场轨道阱质谱对其进行检测。葛根中黄酮类成分,丹参中酚酸类成分及川芎中酚酞类成分分布于不同流分中。从通脉颗粒中初步鉴定了131个化合物,其中葛根59个,丹参26个,川芎43个,丹参和川芎的共有成分3个。26个化合物经过对照品比对鉴定。本研究为中药复方成分的全面分析提供了有效方法。