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| Establishment of a method determining the contents of seven anti-inflammatory active components of the Miao medicine Inula cappa in Raw 264.7 cells |
| ZHOU Jie, XUE Cun, ZHANG Qing, CHEN Siying, CHEN Yi, HUANG Jing, LI Yueting, HUANG Yong, ZHENG Lin, GONG Zipeng |
(School of Pharmacy, Provincial Key Laboratory of Pharmaceutics in Guizhou, State Key Laboratory of Functions and Applications of Medicinal Plants, Guizhou Provincial Engineering Research Center for the Development and Application of Ethnic Medicine and TCM, Guizhou Medical University, Guiyang Guizhou 550004, China)
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Abstract Objective： To establish a method simultaneously determining the contents of seven anti-inflammatory active components including luteolin (LUT), chlorogenic acid (CA), and neochlorogenic acid (NCA), cryptochlorogenic acid (CCA), 3,4-dicaffeoylquinic acid (3,4-DCQA), 3,5-dicaffeoylquinic acid (3,5-DCQA), 4,5-dicaffeoylquinic acid (4,5-DCQA) of the Miao medicine Inula cappa in Raw264.7 cells. Methods： Using mouse macrophage Raw264.7 as the research object, ultra-high liquid chromatography-triple quadrupole mass spectrometry (UPLC-MS/MS) was used to establish the simultaneous determination of LUT, CA, NCA, CCA, 3,4-DCQA, 3,5-DCQA, 4,5-DCQA methods for the content of seven components of the Miao medicine Inula cappa, including specificity, linearity, matrix effect, recovery, accuracy, precision and stability. Results： A rapid and stable analytical method for the measurement of trace drugs in Raw264.7 cells was established. The lowest limit of quantification of the method was 0.48-3.84 ng/mL, and the accuracy was (90.78±5.30)%-(113.99±13.51)%. Its extraction recovery was (87.19±5.33)%-(103.59±14.00)% and the matrix effect was (94.50±15.17)% - (112.77±7.02)%. The stability was (91.71±11.70)%-(109.45±10.63)%, which could meet the measurement requirements. Conclusion： A rapid and accurate method for simultaneous determination content of seven active components LUT, CA, NCA, CCA, 3,4-DCQA, 3,5-DCQA, 4,5-CQA using UPLC-MS/MS was established, which could provide a reference for the study of cellular pharmacokinetics of Inula cappa extract.
［Key words］Inula cappa extract; Raw264.7 cells; anti-inflammatory components； chlorogenic acid; cell pharmacokinetics; UPLC-MS/MS
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Received: 25 March 2022
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