液质联用技术联合化学计量法分析比较续断盐制前后质量差异  

Analysis and comparison of mass differences between raw and salt-processed Dipsaci Radixby LC/MS combined with chemometrics

作　　者：吴杭莎 吕悦 韦飞扬 李昌煜[1,4] 葛卫红 杜伟锋[1,2,3] WU Hang-sha;LYUYue;WEI Fei-yang;LI Chang-yu;GE Wei-hong;DU Wei-feng(School of Pharmaceutical Science,Zhejiang Chinese Medical University,Hangzhou 311402,China;Research Center of Traditional ChineseMedicine Processing Technology,Zhejiang Chinese Medical University,Hangzhou 311401,China;Zhejiang Chinese Medical University Chinese Medicine Yinpian Co.,Ltd.,Hangzhou 311401,China;Academy of Chinese Medical Sciences,Zhejiang Chinese Medical University,Hangzhou 310053,China)

机构地区：[1]浙江中医药大学药学院,浙江杭州311402 [2]浙江中医药大学中药炮制技术研究中心,浙江杭州311401 [3]浙江中医药大学中药饮片有限公司,浙江杭州311401 [4]浙江中医药大学中医药科学院,浙江杭州310053

出　　处：《中草药》2023年第7期2066-2076,共11页Chinese Traditional and Herbal Drugs

基　　金：杭州市农业与社会发展科研重点项目(202204A06);宁波市“科技创新2025”重大专项(20201ZDYF020069);国家重点研发计划(2018YFC1707001);国家中药标准化项目(ZYBZH-H-ZY-45)。

摘　　要：目的通过超高效液相色谱-四级杆-飞行时间质谱技术(UPLC-Q-TOF-MS/MS)鉴定续断DipsaciRadix盐制前后化学成分,并寻找其差异性成分。方法液相采用Agilent Zorbax SB-C18色谱柱(250 mm×4.6 mm,5.0μm),流动相为0.1%甲酸水溶液-乙腈,梯度洗脱,柱温25℃,体积流量0.8 m L/min,检测波长215 nm。质谱采用电喷雾离子源负离子模式采集样品数据,扫描范围为m/z100~2000。根据质谱信息,结合碎片裂解规律,与数据库、文献及对照品的比对对检测到的化学成分进行综合鉴定;通过主成分分析(principal component analysis,PCA)和正交偏最小二乘法-判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)对比分析续断盐制前后的质量差异。结果共鉴定和推测出生续断与盐续断中54种共有化学成分,主要包括皂苷类、环烯醚萜类和酚酸类化合物等。通过多元统计分析表明,生续断和盐续断在PCA模型中能清晰地分为2类,OPLS-DA模型筛选得到12个主要差异成分,分别为马钱苷酸、绿原酸、咖啡酸、马钱苷、异绿原酸B、异绿原酸A、茶茱萸苷、异绿原酸C、续断苷B、续断苷A、川续断皂苷Ⅵ、木通皂苷St(b)。结论建立了生续断与盐续断化学成分定性分析的方法,初步筛选了影响续断盐制前后质量变化的主要化合物,为后续对续断药效物质基础的研究提供了依据,有助于提高续断的质量控制水平。Objective To identify the chemical constituents in Xuduan(Dipsaci Radix)by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry(UPLC-Q-TOF-MS/MS),and find out the differences between raw and salt-processed Dipsaci Radix.MethodsThe liquid phase was performed on an Agilent Zorbax SB-C18column(250 mm×4.6 mm,5.0μm),the mobile phase was 0.1%aqueous formic acid-acetonitrile,gradient elution,the column temperature was 25℃,the flow rate was 0.8 mL/min,and the detection wavelength was 215 nm.The mass spectrometry used electrospray ion source negative ion mode to collect sample data,and the scanning range was m/z100-2000.According to mass spectrometry information,combined with fragmentation rules,the detected chemical components were comprehensively identified by comparing with databases,literature and reference substances.Then,the quality differences before and after the salting process were compared and analyzed by principal component analysis(PCA)and orthogonal partial least squares-discriminant analysis(OPLS-DA).ResultsA total of 54 common chemical constituents were identified and speculated in raw and salt-processed Dipsaci Radix,mainly including saponins,iridoids and phenolic acids.Through multivariate statistical analysis,it was shown that the raw and salt-processed Dipsaci Radixcould be clearly divided into two categories in the PCA model.At the same time,there were 12 main differential components,includingloganic acid,chlorogenic acid,caffeic acid,loganin,isochlorogenic acid B,isochlorogenic acid A,cantleyoside,isochlorogenic acid C,dipsanoside B,dipsanoside A,asperosaponin VI,Akebiasaponin St(b),had been screened by OPLS-DA model.ConclusionIn this study,a method for qualitative analysis of the chemical components of was established to screen the main material basis affecting the quality change raw and salt-processed Dipsaci Radixpreliminarily.It provideda basis for subsequent studies on the material basis of the medicinal effect of the Dipsaci Radixand helpedto improve the

关 键 词：续断 盐制 UPLC-Q-TOF-MS/MS 质量差异 化学计量法 主成分分析 正交偏最小二乘法-判别分析 皂苷 环烯醚萜 酚酸 马钱苷酸 绿原酸 咖啡酸 马钱苷 异绿原酸B 异绿原酸A 茶茱萸苷 异绿原酸C 续断苷B 续断苷A 川续断皂苷Ⅵ 木通皂苷St(b) 

分 类 号：R283.6[医药卫生—中药学]