UPLC-Q-TOF-MS/MS快速鉴定补肺健脾方的化学成分  

Identification of Chemical Constituents in Bufei Jianpi Formula by UPLC-Q-TOF-MS/MS

作　　者：崔琳琳 包永睿[1] 王帅[1] 李天娇[1] 孟宪生[1] CUI Lin-lin;BAO Yong-rui;WANG Shuai;LI Tian-jiao;MENG Xian-sheng(School of Pharmacy,Component Medicine Engineering Research Center of Liaoning Province,Liaoning Province Modern Chinese Medicine Research Engineerig Laboratory,Liaoning University of Traditional Chinese Medicine,Dalian 116600,China)

机构地区：[1]辽宁中医药大学药学院,辽宁省组分中药工程技术研究中心,辽宁省现代中药研究工程实验室,辽宁大连116600

出　　处：《中国实验方剂学杂志》2020年第9期184-193,共10页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家重点研发计划项目(2018YFC1704805);辽宁省中医药现代化研究创新团队项目(LT2017015)。

摘　　要：目的:采用UPLC-Q-TOF-MS/MS技术对补肺健脾方中的化学成分进行检测和分析。方法:采用Agilent Poroshell SB-C18色谱柱(4.6 mm×100 mm,2.7μm),正、负离子模式下的流动相系统分别选择0.1%甲酸水溶液(A)-乙腈(B)梯度洗脱(0~10 min,3%B;10~100 min,3%~50%B;100~120 min,50%~100%B)和水(A)-乙腈(B)梯度洗脱(0~5 min,3%B;5~60 min,3%~100%B),流速0.6 mL·min^-1,柱温30℃。采用电喷雾离子源(ESI),干燥气流速13 L·min^-1,干燥气温度350℃,毛细管电压3.5 kV,雾化器压力310.28 kPa,碎裂电压125 V,二级质谱碰撞电压40 eV,在正、负离子扫描采集数据,采集范围均为m/z 50~1000,通过Agilent MassHunter Qualitative Analysis数据分析定性软件进行色谱峰的提取和匹配。结果:通过质谱信息并结合对照品、相关文献、数据库检索,共鉴别出了95个化合物,包括黄酮类41个,生物碱类23个、木脂素类12个,有机酸类9个和其他类化合物10个。结论:补肺健脾方化学成分复杂,不同成分裂解规律各异,黄酮类类化合物在裂解过程中容易发生脱糖基、脱水、环的逆狄尔斯-阿尔德反应(RDA)裂解,以及CO,CO2,CHO等一些中性分子丢失;木脂素类成分苯环上常有羟基、羰基、甲氧基等取代基,易得到丢失H2O或CO的碎片离子;有机酸类化合物的基本结构为酚羟基取代的芳香环、丙烯酸、脂肪酸,该类化合物在负离子模式下易丢失H2O和COOH,且易在羰基处断裂形成碎片离子。该方法快速、灵敏、准确,可快速鉴别补肺健脾方的化学成分,为阐释该复方的功效物质基础提供了依据。Objective:To identify and analyze the chemical constituents in Bufei Jianpi formula by UPLCQ-TOF-MS/MS.Method:An Agilent Poroshell SB-C18 column(4.6 mm×100 mm,2.7μm)was used with a mobile phase system of 0.1%formic acid solution(A)-acetonitrile(B)for gradient elution(0-10 min,3%B;10-100 min,3%-50%B;100-120 min,50%-100%B)under positive ion mode and water(A)-acetonitrile(B)for gradient elution(0-5 min,3%B;5-60 min,3%-100%B)under negative ion mode,the flow rate was 0.6 mL·min-1,and the column temperature was 30℃.Mass spectrometric data were obtained under electrospray inoization(ESI)in positive and negative ion modes,the collection range was m/z 50-1000.Agilent MassHunter Qualitative Analysis software was used to extract and match chromatographic peaks.Result:Combined with reference,related literature and database analysis,95 compounds were identified by mass spectrometry information,including 41 flavonoids,23 alkaloids,12 lignans,9 organic acids,and 10 other compounds.Conclusion:The chemical composition of Bufei Jianpi formula is complex,and the cracking rules of different components are different.Flavonoids are prone to deglycosylation,dehydration,Diels-Alder reaction(RDA)cleavage of the ring during lysis,and loss of some neutral molecules such as CO,CO2,CHO.Lignans has a substituent such as a hydroxyl group,a carbonyl group or a methoxy group on the benzene ring,and it is easy to obtain a fragment ion which loses H2O or CO.The basic structure of organic acids is a phenolic hydroxyl groupsubstituted aromatic ring,acrylic acid,fatty acid or the like,this kind of compound is easy to lose H2O and COOH in negative ion mode,and it is easy to break at the carbonyl to form fragment ions.This established method is rapid,sensitive and accurate,which can quickly identify the chemical constituents in Bufei Jianpi formula and provide evidences for clarifying efficacy material base of this formula.

关 键 词：补肺健脾方 化学成分 黄酮类 生物碱类 木脂素类 有机酸类 慢性阻塞性肺疾病 

分 类 号：R22[医药卫生—中医基础理论] R914[医药卫生—中医学]