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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).
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.
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.
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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| 科 Family | 属数 Number of genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) | 属 Genus | 种数 Number of species | 占总种数比例 Percentage of total species (%) |
|---|---|---|---|---|---|---|
| 鹅膏菌科Amanitaceae | 2 | 11 | 5.26 | 鹅膏菌属 Amanita | 10 | 4.78 |
| 小菇科 Mycenaceae | 2 | 12 | 5.74 | 丝盖伞属 Inocybe | 5 | 2.39 |
| 多孔菌科 Polyporaceae | 8 | 14 | 6.70 | 蜡蘑属 Laccaria | 5 | 2.39 |
| 红菇科 Russulaceae | 3 | 23 | 11.00 | 小皮伞属 Marasmius | 6 | 2.87 |
| 小菇属 Mycena | 11 | 5.26 | ||||
| 光柄菇属 Pluteus | 5 | 2.39 | ||||
| 红菇属 Russula | 17 | 8.13 | ||||
| 栓菌属 Trametes | 5 | 2.39 |
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