Screening of Anti-EGFR Components from Aconitum Soongaricum Stapf. in Xinjiang Province Based on Cell Membrane Chromatography / 中国药师

Objective:To screen anti-EGFR components from Aconitum soongaricum Stapf. and investigate their inhibitory effect on HEK293 cells and EGFR/HEK293 high expression cell lines. Methods:A two-dimensional online system (EGFR/CMC-HPLC-MS) was built to screen anti-EGFR active compounds from Aconitum soongaricum Stapf.;molecular docking assay was performed to determine the binding affinity of the components to EGFR;MTT was used to confirm the inhibition effect of the screened compounds on HEK293 cells and EGFR/HEK293 high expression cell lines.Results:The screening results showed that aconitine,12-epi-napellin and songorine were the active components acting on EGFR like gefitinib; molecular docking assay showed the targeting components acting on EGFR in the similar manner with gefitinib used as a positive control drug; MTT assay confirmed the screened components had inhibitory effects on HEK293 cells and EGFR/HEK293 high expression cell lines in a dose-dependent manner within the dose range of 0.4- 50.0 μmol·L-1. Conclusion:The screening results are apparently consistent with the pharmacological effect. The online EGFR/CMC-HPLC-MS method has a good application prospect in the rapid screening of the active compounds in the complex system of traditional Chinese medicines,and fully demonstrates such advantages of cell membrane chromatography as simple and rapid with high efficiency.

Optimization of Extraction Process of Yinhuai Jiedu Granula by Response Surface Methodology / 中国中医药信息杂志

Objective To optimize extraction process of Yinhuai Jiedu Granula.Methods On the basis of single factor experiments, effects of soaking time, solvent dosage and extracting time on contents of chlorogenic acid and total flavonoids were investigated by response surface methodology. HPLC was used to determine contents of chlorogenie acid and total flavonoids with mobile phase of acetonitrile-0.1% phosphoric acid for gradient elution (0–20 min, 13%A; 20–30 min, 13%–18%A; 30–55 min, 18%–20%A; 55–65 min, 20%–23%A) and detection wavelengthes of 327 nm and 256 nm for chlorogenic acid and total flavonoids respectively. Results Optimal extraction process was as following: soaked 60 minutes and extracted twice with 12 times the amount of water per time of 130 minutes. Concentrations of chlorogenic acid, rutin and baicalin were 51.497, 68.872, 25.763 μg/g respectively. The average extract yield was 33.105%. Conclusion The optimized extraction process is feasible and stable,which can provide data support for the further research on Yinhuaijiedu granula.