基于UPLC-Q-TOF/MS;分析不同炮制时间炒车前子的化学成分变化规律

Analysis of Chemical Composition Variations in Plantaginis Semen at Different Frying Time Based on UPLC-Q-TOF/MS

目的:为探究不同炮制时间炒车前子化学成分变化情况,采用超高效液相色谱-四极杆-飞行时间质谱法(UPLCQ-TOF/MSE)对不同炒制时间炒车前子中差异成分进行快速鉴定和半定量分析。方法:色谱条件采用Waters ACQUITY UPLC BEH C_(18)色谱柱(2.1 mm×100 mm,1.8μm),流动相0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~1 min,5%~10%B;1~2 min,10%~15%B;2~10 min,15%~20%B;10~12 min,20%~40%B;12~13 min,40%~100%B;13~14 min,100%~5%B;14~15 min,5%B),流速0.3 mL·min^(-1),柱温40℃,进样量3μL。质谱分析使用电喷雾离子源(ESI),正、负离子模式检测,扫描范围为m/z50~1500。采用MarkerLynx 4.1软件挖掘差异化合物,比较不同炒制时间样品各离子峰的离子响应强度,研究化学成分含量变化情况。结果:共找到具有潜在的显著性差异成分20个,其中17个成分已鉴定,3个成分未知,主要包括苯乙醇苷类、环烯醚萜苷类、生物碱类等。炮制后蔗糖、京尼平苷酸、毛蕊花糖苷、车前素A等7个成分峰强度降低;orobanchoside,dianthoside和车前草苷D峰强度在炒制过程中先升高后降低;去咖啡酰基毛蕊花糖苷,木通苯乙醇苷A,异毛蕊花糖苷等10个成分峰强度升高,其中9个为新生成成分。结论:炒制后,炒车前子化学成分的含量和组成均发生了显著变化,这可能与车前子炒制后通便作用减弱、止泻利尿活性增强有关。

Objective:In order to explore the changes of chemical constituents in Plantaginis Semen before and after stir-frying,ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF/MSE)was used to rapidly identify and semi-quantitatively analyze the differential components in Plantaginis Semen processed at different stir-frying time.Method:Waters ACQUITY UPLC BEH C_(18)column(2.1 mm×100 mm,1.8μm)was employed with the mobile phase of 0.1%formic acid aqueous solution(A)-acetonitrile(B)for gradient elution(0-1 min,5%-10%B;1-2 min,10%-15%B;2-10 min,15%-20%B;10-12 min,20%-40%B;12-13 min,40%-100%B;13-14 min,100%-5%B;14-15 min,5%B),the flow rate was 0.3 mL·min^(-1),the column temperature was 40℃,and the injection volume was 3μL.Electrospray ionization(ESI)was applied for mass spectrometric analysis under positive and negative ion modes,and the scanning range was m/z 50-1500.MarkerLynx 4.1 software was used to find the differential compounds,and the intensity of each ion peak in samples with different stir-frying time was compared to study the content variations of these compounds.Result:A total of 20 components with potential significant differences were found,among which 17 were identified and 3 were unknown,mainly including phenylethanoid glycosides,iridoid glycosides,alkaloids and others.After processing,the peak intensities of 7 compounds,such as sucrose,geniposidic acid,verbascoside and plantagoguanidinic acid A,in Plantaginis Semen decreased.The peak intensities of orobanchoside,dianthoside and plantain D increased first and then decreased during the stirfrying process.The peak intensities of 10 compounds(decaffeoylacteoside,calceolarioside A,isoacteoside,etc.)increased,and 9 of them were newly generated components.Conclusion:The content and composition of the chemical components in Plantaginis Semen changed significantly after stir-frying,which may be related to the reduction of laxative effect and the enhancement of antidiarrheal and diuretic activities of Plantaginis Semen after stir-frying.