基于UPLC-Q-TOF-MS谱-效分析的浙贝母化痰质量标志物的初步筛选及含量差异研究  

Quality Markers Screening of Phlegm-eliminating Effect of Fritillaria thunbergii and Their Content Differences by UPLC-Q-TOF-MS Spectrum-Effect Analysis Technology

作　　者：程斌[1] 童静玲 周爱珍[1] 阮洪生[1] 王昕蕾 彭昕 CHENG Bin;TONG Jing-ling;ZHOU Ai-zhen;RUAN Hong-sheng;WANG Xin-lei;PENG Xin(Traditional Chinese Medicine College,Zhejiang Pharmaceutical College,Ningbo 315100,China;Ningbo Research Institute of Zhejiang University,Ningbo 315000,China;College of Pharmacy,Harbin University of Commerce,Harbin 150076,China;Luqiao Hospital of Taizhou Medical Center in Zhejiang,Taizhou 318050,China)

机构地区：[1]浙江医药高等专科学校中药学院,浙江宁波315100 [2]浙江大学宁波研究院,浙江宁波315000 [3]哈尔滨商业大学药学院,哈尔滨150076 [4]台州恩泽医疗中心(集团)恩泽医院,浙江台州318050

出　　处：《中国药学杂志》2021年第6期462-471,共10页Chinese Pharmaceutical Journal

基　　金：浙江省公益性技术应用研究计划资助(LGN19H280002);浙江省食品药品监督管理局2018年度科技计划项目资助(2018018);宁波市科技创新2025重大专项资助(2019B10008);2020年度高等学校国内访问工程师“校企合作项目”资助(FG2020003)。

摘　　要：目的探讨10种不同产地规格浙贝母超高效液相色谱-四极杆-飞行时间质谱(UPLC-Q-TOF-MS)指纹谱与其化痰作用的谱-效关系,初步筛选浙贝母化痰作用质量标志物(Q-marker),并比较主要质量标志物的含量差异。方法应用UPLC-Q-TOF-MS技术建立浙贝母指纹图谱。液相条件:色谱柱ZORBAX HPLC Eclipse Plus C18(3 mm×150 mm,1.8μm),以乙腈-0.1%甲酸水为流动相进行梯度洗脱,流速0.4 mL·min^(-1);柱温30℃;进样量2μL。质谱条件:通过运用电喷雾(ESI)离子源,在正负离子模式下进行数据采集。采用小鼠酚红法以气管酚红排泌量为指标评价浙贝母化痰作用。结合双变量相关分析和灰色关联度分析研究浙贝母指纹图谱中共有峰与化痰作用的相关性,筛选出关联性较大的成分作为其化痰作用的主要质量标志物。采用高效液相色谱-蒸发光散射检测(HPLC-ELSD)建立其中4个主要成分的含量测定方法,并比较不同产地规格的含量差异。结果初步筛选出6个与浙贝母化痰作用相关的质量标志物,分别为腺苷、贝母辛,西贝素、贝母素乙、贝母素甲、浙贝素。10个不同产地规格浙贝母中贝母辛含量在0.186~1.1069 mg·g^(-1)内;西贝素含量在0.2047~1.2439 mg·g^(-1)内;贝母素乙含量在0.0842~0.6115 mg·g^(-1)内;贝母素甲含量在1.4585~11.7696 mg·g^(-1)内。结论基于“谱-效”关联的质量标志物研究策略是一种快速分析及初步确定浙贝母化痰作用药效物质的有效方法。在此基础上建立的浙贝母主要质量标志物HPLC-ELSD含量测定的方法可靠、准确。本实验对完善浙贝母质量控制方法,制定科学的浙贝母商品标准规范具有较重要的参考价值。OBJECTIVE To explore the relationship between the UPLC-Q-TOF-MS fingerprints of Fritillaria thunbergii and its phlegm-reducing effect in 10 different producing areas,screen the quality markers of phlegm-removing phlegm of Fritillaria thunbergii,and compare the content of main components.METHODS UPLC-Q-TOF-MS technology was used to establish the fingerprint of Fritillaria thunbergii.Liquid conditions:chromatographic column ZORBAXHPLC Eclipse Plus C18(3 mm×150 mm,1.8μm),gradient elution with methanol-0.1%formic acid water as mobile phase,flow rate was 0.4 mL·min^(-1);column temperature was maintained at 30℃;injection volume was 2μL.Mass spectrometry conditions:an electrospray(ESI)ion source was used to collect data in positive and negative ion mode.The mouse phenol red method was used to evaluate the phlegm-reducing effect of Fritillaria thunbergii with tracheal phenol red excretion.Combining bivariate correlation analysis and gray correlation analysis to study the correlation between the common peak in the fingerprint of Fritillaria thunbergii and the phlegm effect,and screen out the components with greater correlation as quality markers for its phlegm effect,and used HPLC-ELSD method established the determination method for the content of 4 main components.RESULTS Six quality markers related to the phlegm-reducing effect of Fritillaria thunbergii were screened out:adenosine,peimisine,imperialine,peiminine,peimine,and zhebeine.The content of peimisine was in the range of 0.186-1.1069 mg·g^(-1);the content of imperialine was in the range of 0.2047-1.2439 mg·g^(-1);the content of peiminine was in the range of 0.0842-0.6115 mg·g^(-1);the content of peimin was in 1.4585-11.7696 mg·g^(-1).CONCLUSION The research strategy of quality markers based on"spectrum-effect"correlation is an effective method for rapid analysis and preliminary determination of the effective components of Fritillaria thunbergii Miq.in resolving phlegm.On this basis,the established HPLC-ELSD method for determination of the main quality ma

关 键 词：浙贝母 谱效关系 超高效液相色谱-四极杆-飞行时间质谱 高效液相色谱-蒸发光散射检测 质量标志物 

分 类 号：R917[医药卫生—药物分析学]