Activity of anti-epidermal growth factor receptor in Calla Chinensis and UPLC/Q-TOF-MS analysis on its effective part / 中草药

ABSTRACT

Objective: Using homogeneous time-resolved fluorescence (HTRF) technology to screen the Calla Chinensis extracts with anti-epidermal growth factor receptor activity and to analyze the active ingredients in Calla Chinensis by ultra-performance liquid chromatography/quadrupole-time of flight mass-spectrometry (UPLC/Q-TOF-MS). Methods: After percolation with petroleum ether, ethanol extract, ethyl acetate extraction, and boiling with water, four fractions were obtained. HTRF method was applied to detecting the inhibition of ethyl acetate fraction on EGFR, and the inhibitory rate was calculated. The chromatographic separation was performed on Acquity UPLC BEH C18 column with a gradient elufion of 0.1% formic acid water-acetonitrile. The mass spectrometer equipped with electrospay ionization source was used as defector, data were collected under the positive ion modes, and screened at 200-400 nm. Results: The ethyl acetate fraction of Calla Chinensis showed the strong inhibitory activity on EGFR. Fourteen compounds were analyzed and identified, among which tannins were the main active components, the 11 tannin compounds were punicalin, vanillin, di-HHDP-glucose, ellagitannin, myricetin 3-O-rhamnoside, protocatechuic acid, gallotannin-glucose, ellagic acid-hexose, 3, 5-dicaffeoylquinic acid, monogalloyl-glucose, and gallotannin, and the three others were unknown. Conclusion: The ethyl acetate fraction of Calla Chinensis has the significant EGFR inhibitory activity, and the IC50 value is 5.528 μg/mL. Tannins, including Chinese tannin, gallogen, and ellagitannin as main constituents, are identified through the information of positive ion and relative molecular mass determined by Q-TOF-MS. The results indicate that Calla Chinensis contains tannin ingredients to inhibit EGFR, in the hope to provide a theoretical basis of the application in anticancer and to lay the foundation for the further tracking separation of the active ingredients.