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.

Detection of minor constituents in TongMai Granules using NP×RP off-line twodimensional liquid chromatography coupled with orbitrap mass spectrometry

TongMai Granules is a Chinese medicine patent drug to treat cardiovascular diseases.It is prepared from the water extract of Dan-shen,Chuan-xiong and Ge-gen.The main compounds of TongMai Granules are isoflavone glycosides,isoflavones,and phenolic acids.However,it is difficult to detect low-polar compounds such as tanshinones and phthalides though they may also contribute to the therapeutic effect.In this work,an off-line two-dimensional chromatography method was developed to enrich the minor constituents in TongMai Granules.The extract was subjected to an SEP-PAK silica SPE column and eluted with petroleum ether,ethyl acetate–methanol(6:1,v/v),and methanol,successively.The three fractions were then analyzed using ultra-high performance liquid chromatography coupled with orbitrap mass spectrometry(UHPLC/orbitrap-MS).The isoflavones from Ge-gen,phenolic acids from Dan-shen and phthalides from Chuan-xiong were distributed in different fractions.A total of 131 compounds,including 59 from Ge-gen,26 from Dan-shen,43 from Chuan-xiong,and 3 common ones from Dan-shen and Chuan-xiong were tentatively identified.Among them,26 compounds were identified by comparing with reference standards.This study set a good example for comprehensive analysis of the constituents in traditional Chinese medicine formula.