太白茶HPLC指纹图谱的建立及UHPLC-Q Exactive 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
李莎莎 1李雅娟 2王晓婷 1毛阿娟 1李凡 1李芳 1张红 1王卫锋1
作者信息
- 1. 陕西省中医药研究院,西安 710003
- 2. 陕西中医药大学,咸阳 712046
- 折叠
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,5 μm)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10 μL.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,1.7 μm)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 80～1 200.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.
Abstract
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-C18(250 mm × 4.6 mm,5 μm)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 10 μL.HPLC fingerprints of lichens of Thamnolia subuliformis was established.The soft-ware 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 C18(50 mm ×2.1 mm,1.7 μm)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-1 and the column temperature was 30 ℃.Mass spec-trometry was performed using an electrospray ionization and data was collected in negative ion modes in the range of m/z 80-1 200.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 estab-lished 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Key words
Thamnolia subuliformis(Ehrh.)W.Culb./HPLC/fingerprinting/quality evaluation/UHPLC-Q Exactive Focus MS/MS/qualitative analysis引用本文复制引用
基金项目
陕西省中医药局中医药全省性专款专项(2021-QYZL-01)
陕西省中医药研究院苗圃培育计划(2021-16)
陕西省中医药研究院苗圃培育计划(2021-21)
陕西省中医药局传承创新"秦药"开发重点科学研究项目(2021-02-GJ-021)
陕西省重点研发计划一般项目(2022SF-301)
陕西省重点研发计划一般项目(2024SF-YBXM-481)
出版年
2024
NETL
NSTL
万方数据