太白茶HPLC指纹图谱的建立及UHPLC-QExactive Focus MS/MS联用分析  

Eatablishment of HPLC fingerprint of lichens of Thamnolia subuliformis(Ehrh.)W.Culb.and its UHPLC-Q Exactive Focus MS/MS analysis

作　　者：李莎莎 李雅娟 王晓婷 毛阿娟 李凡[1] 李芳[1] 张红[1] 王卫锋[1] LI Sha-sha;LI Ya-juan;WANG Xiao-ting;MAO A-juan;LI Fan;LI Fang;ZHANG Hong;WANG Wei-feng(Shaanxi Provincial Academy of Traditional Chinese Medicine,Xi’an 710003,China;Shaanxi University of Chinese Medicine,Xianyang 712046,China)

机构地区：[1]陕西省中医药研究院,西安710003 [2]陕西中医药大学,咸阳712046

出　　处：《药物分析杂志》2024年第4期585-593,共9页Chinese Journal of Pharmaceutical Analysis

基　　金：陕西省中医药管理局中医药全省性专款专项项目(2021-QYZL-01);陕西省中医药管理局传承创新暨“秦药”开发重点科学研究项目(2021-02-GJ-021);陕西省中医药研究院“苗圃培育计划”项目(2021-16,2021-21);陕西省重点研发计划一般项目(2022SF-301,2024SF-YBXM-481)。

摘　　要：目的:建立太白茶药材高效液相色谱(HPLC)指纹图谱分析方法,并运用超高效液相色谱-四极杆/静电场轨道阱高分辨质谱(UHPLC-Q Exactive Focus MS/MS)法对太白茶中的特征峰进行定性分析。方法:采用Agilent TC-C18(250 mm×4.6 mm,5μm)色谱柱,以甲醇(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱,流速1.0 mL·min^(-1),柱温30℃,检测波长254 nm,进样量10μL。采用“中药色谱指纹图谱相似度评价系统2012年版”,建立指纹图谱并进行相似度评价。化学成分分析选用UHPLC-Q Exactive Focus MS/MS技术,采用Waters Acquity UPLC BEH C18(50 mm×2.1 mm,1.7μm)色谱柱,以乙腈(A)-0.1%甲酸水(B)为流动相,梯度洗脱,流速0.3 mL·min^(-1),柱温30℃;质谱条件为电喷雾离子源(ESI),负离子模式扫描,扫描范围m/z 80~1200。结果:建立的太白茶特征指纹图谱中,标定共有峰8个,10批太白茶药材相似度均≥0.9,各批次之间共有峰的相对保留时间RSD均<0.5%。对8个共有峰中的4个峰进行了结构指认,依次为鳞片衣酸、羊角衣酸、thamnoliadepsides B、坝巴酸。结论:所建立的HPLC指纹图谱方法稳定可靠,专属性好,结合UHPLC-Q-Exactive Focus-MS/MS的定性分析,可用于太白茶药材的整体质量综合评价。Objective:To establish a quality assessment method for lichens of Thamnolia subuliformis(Ehrh.)W.Culb.based on HPLC fingerprint,and qualitative analysis of the chemical constituents by ultra-high performance liquid chromatography coupled with quadrupole/electrostatic field orbital trap high resolution mass spectrometry(UHPLC-Q Exactive Focus MS/MS).Methods:Agilent TC-Cis(250 mm×4.6 mm,5μm)chromatographic column was used,the mobile phase was methol-0.1%phosphoric acid with gradient elution at the flow rate of 1.0 mL·min^(-1),the column temperature was 30℃,the detection wavelength was 254 nm,and the injection volume was 10μL.HPLC fingerprints of lichens of Thamnolia subuliformis was established.The software of similarity calculation for traditional Chinese medicines fingerprints(version 2012)was used to establish the fingerprinting of lichens of Thamnolia subuliformis.The chemical constituents were analyzed by UHPLC-Q Exactive Focus MS/MS,The chromatographic separation was performed on a Waters Acquity UPLC BEH Cis(50 mm×2.1 mm,1.7μm)column with acetonitrile(A)-0.1%formic acid aqueous solution(B)as mobile phase for gradient elution,the flow rate was 0.3 mL·min*'and the column temperature was 30℃.Mass spectrometry was performed using an electrospray ionization and data was collected in negative ion modes in the range of m/z 80-1200.Results:Eight common peaks were identified from fingerprints of 10 batches of samples.The RSD values of relative retention time of 8 common peaks of chromotograms of samples were all below 0.5%and the similarities were above 0.9.4 of the identified peaks were further confirmed by UHPLC-Q Exactive Focus MS/MS as squamatic acid,baeomycesic acid,thamnoliadepsides B,barbatinic acid.Conclusion:The established method of fingerprint is stable,reliable,and specific,which can be used for quality evaluation of lichens of Thamnolia subuliformis.

关 键 词：太白茶 高效液相色谱 指纹图谱 质量评价 超高效液相色谱-四极杆/静电场轨道阱高分辨质谱 定性分析 

分 类 号：R917[医药卫生—药物分析学]