基于UPLC-Q-TOF-MS技术的黄芪桂枝五物汤基准样品化学成分分析及表征

Chemical Composition Analysis and Characterization of Reference Sample of Huangqi Guizhi Wuwutang Based on UPLC-Q-TOF-MS

目的:对黄芪桂枝五物汤基准样品(冻干粉)进行化学成分分析,为该经典名方制剂的质量标准制定提供质量标志物。方法:采用超高效液相色谱-四级杆-飞行时间-串联质谱法(UPLC-Q-TOF-MS),色谱条件为Waters ACQUITY UPLC?HSS T3色谱柱(2.1 mm×100 mm,1.8μm),流动相甲醇(A)-0.1%甲酸水溶液(B)梯度洗脱(0~8 min,1%~20%A;8~10 min,20%~30%A;10~12 min,30%~35%A;12~14 min,35%~40%A,14~23 min,40%~55%A,23~27 min,55%~99%A;27~28 min,99%A;28~28.5 min,99%~1%A;28.5~30 min,1%A),柱温40℃,进样量2μL,流速0.3 mL·min^(-1)。在正、负离子模式下采集黄芪桂枝五物汤基准样品(冻干粉)的质谱数据,质谱条件为电喷雾离子源(ESI),扫描范围m/z 50~1200,碰撞能量10~30 eV。利用UNIFI 1.8和Progenesis QI 2.0软件解析,结合对照品和文献信息比对,对黄芪桂枝五物汤基准样品(冻干粉)的化学成分进行表征。结果:一共表征出123化学成分,其中黄酮类33个、苷类26个、有机酸类18个、萜类11个、苯丙素类7个、姜辣素类4个、生物碱类3个、氨基酸类3个、酰胺类2个、其他类16个。结论:建立的方法可以快速、准确地表征黄芪桂枝五物汤基准样品(冻干粉)中的化学成分,可为该经典名方的质量评价指标筛选提供依据,并为其制剂研究提供参考。

Objective:To analyze the chemical composition of the reference sample of Huangqi Guizhi Wuwutang(lyophilized powder),and to provide quality markers for the formulation of quality standards of this formula.Method:Ultra performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry(UPLC-Q-TOF-MS)was performed on a Waters ACQUITY UPLC?HSS T3 column(2.1 mm×100 mm,1.8μm),the mobile phase was methanol(A)-0.1%formic acid aqueous solution(B)for gradient elution(0-8 min,1%-20%A;8-10 min,20%-30%A;10-12 min,30%-35%A;12-14 min,35%-40%A,14-23 min,40%-55%A,23-27 min,55%-99%A;27-28 min,99%A;28-28.5 min,99%-1%A;28.5-30 min,1%A),the column temperature was 40℃,the injection volume was 2μL,and the flow rate was 0.3 mL·min^(-1).The mass spectrometry data of the reference sample of Huangqi Guizhi Wuwutang(lyophilized powder)were collected under positive and negative ion modes.The conditions of mass spectrometry were electrospray ionization(ESI),scanning range of m/z 50-1200,and impact energy of 10-30 eV.UNIFI 1.8 and Progenesis QI2.0 software were used to analyze and characterize the chemical constituents in reference sample of Huangqi Guizhi Wuwutang(lyophilized powder)combined with reference comparison and literature review.Result:A total of 123 chemical constituents were identified,including 33 flavonoids,26 glycosides,18 organic acids,11 terpenoids,7 phenylpropanoids,4 gingerol,3 alkaloids,3 amino acids,2 amides and 16 other compounds.Conclusion:The established method can quickly and accurately characterize the chemical components in the reference sample of Huangqi Guizhi Wuwutang(lyophilized powder),which can provide a basis for the selection of quality evaluation indicators of this formula,and provide a reference for its preparation research.