目的: 基于超高效液相色谱串联四极杆飞行时间质谱联用(UPLC-Q TOF MS/MS)技术,系统表征地胆草中化学成分并同时测定其中13个化学成分(新绿原酸、绿原酸、隐绿原酸、木犀草素-7-O-葡萄糖醛酸苷、木犀草苷、异绿原酸B、异绿原酸A、芹菜素-7-O-β-D-葡萄糖醛酸苷、异绿原酸C、去氧地胆草素、异去氧地胆草素、异地胆草种内酯、地胆草种内酯)的含量。方法: 采用UPLC-Q TOF MS/MS技术,结合顺序窗口采集所有理论质谱模式(SWATH),采集不同批次地胆草样品的数据。使用Waters HSS T3 C18(100 mm×2.1 mm,1.8 μm)色谱柱,以乙腈(A)-0.1%甲酸水溶液(B)为流动相,梯度洗脱,流速0.4 mL·min-1,柱温35 ℃,进样量2 μL。采用电喷雾离子源(ESI),正、负离子模式下采集数据。结合对照品信息,参考相关文献完成对地胆草中化学成分的结构鉴定。同时建立地胆草中13个化学成分的含量测定方法并进行方法学考察。结果: 地胆草中初步鉴定出36个化学成分,其中包括有机酸类20个,黄酮类7个,倍半萜内酯类6个,香豆素类1个及其他成分2个;含量测定的13个成分在测定浓度范围内线性关系良好,相关系数≥0.999 0,精密度、稳定性、重复性等均符合要求,平均加样回收率为94.9%~103.5%,RSD≤3.8%。不同批次地胆草所含化学成分基本相似,但13个成分的含量存在一定差异。结论: 本研究建立的UPLC-Q TOF MS/MS方法,可同时对地胆草进行定性定量分析,方法灵敏准确,稳定性和重复性良好,可以为地胆草的质量控制及开发提供方法参考及数据支撑。
果佳慧, 蔡于罗, 田伟, 高乐, 李葆林, 甄亚钦, 牛丽颖
. 基于UPLC-Q TOF MS/MS技术结合SWATH采集法同时定性定量分析地胆草中的化学成分*[J]. 药物分析杂志, 2024
, 44(8)
: 1331
-1342
.
DOI: 10.16155/j.0254-1793.2023-0470
Objective: To systematically characterize and simultaneous determine 13 chemical components (neochlorogenic acid, chlorogenic acid, cryptochlorogenic acid, luteolin-7-glucuronide, cynaroside, isochlorogenic acid B, isochlorogenic acid A, apigenin-7-glucuronide, isochlorogenic acid C, deoxyelephantopin, isodeoxyelephantopin, isoscabertopin and scabertopin) in Elephantopi Herba based on ultra performance liquid chromatography-quadrupole-time-of-flight tandem mass spectrometry (UPLC-Q TOF MS/MS). Methods: UPLC-Q TOF MS/MS technology combined with sequential window acquisition of all theoretical mass spectra (SWATH) mode was used to collect data of different batches of Elephantopi Herba samples. The separation was performed on a Waters HSS T3 C18(100 mm×2.1 mm, 1.8 μm) column with mobile phase consisting of acetonitrile and 0.1% formic acid water solution. The flow rate of gradient elution was 0.4 mL·min-1, the column temperature was 35 ℃ and the injection volume was 2 μL. The electrospray ionization (ESI) source was employed to collect data under both positive and negative ion modes. With reference to reference substances and relevant literatures, chemical components in Elephantopi Herba were identified. At the same time, the determination method for 13 chemical components in Elephantopi Herba was established and validated. Results: In this study, the cleavage patterns of different classes of compounds were summarized, and 36 chemical components were initially identified, including 20 organic acids, 7 flavonoids, 6 sesquiterpene lactones, 1 coumarin and 2 other components. All of the analytes showed good linearity (r≥0.999 0) in the tested ranges. The precision, repeatability and stability of the method were good for the 13 components. The average recoveries were in the range of 94.9%-103.5% with relative standard deviations (RSDs) ≤3.8%. The chemical compositions of different batches of Elephantopi Herba were basically similar, but there were certain differences in the contents of 13 components. Conclusion: In this study, comprehensive characterization of the chemical constituents and relative quantitative analysis of Elephantopi Herba are carried out, and a rapid and efficient qualitative analysis method of Elephantopi Herba is established by UPLC-Q TOF MS/MS, which provides a reference for the screening and rational exploitation of the medicinal substances of Elephantopi Herba, and methodological reference and data support for the quality control and development of Elephantopi Herba.
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