ABSTRACT
OBJECTIVE：To study the spectrum-effect relationship of HPLC finger print of different polar parts of Ampelopsis grossedentata with its in vivo anti-inflammatory effect. METHODS ：A. grossedentata was reflux extracted with 70％ ethanol，then extracted with petroleum ether ，chloroform，ethyl acetate and water saturated n-butanol；or it was directly decocted with water and then concentrated to obtain different polar parts. The xylene-induced mice ear swelling model was established ；using dexamethasone as positive control ，anti-inflammatory activity of different polar parts of A. grossedentata was investigated. Fingerprints of different polar parts of A. grossedentata were established by HPLC. The determination was performed on Poroshell 120 EC-C18 column with mobile phase consisted of acetonitrile- 0.1％ phosphoric acid solution （gradient elution ）at the flow rate of 1 mL/min. The column temperature was 25 ℃. The detection wavelength was set at 365 nm，and sample size was 5 μL. The grey ralational analysis method was used to analyze the spectrum-effect relationship of HPLC fingerprint common peaks of different polar parts of A. grossedentata with its anti-inflammatory effect. The correlation coefficient and correlation degree were calculated and ranked. RESULTS：Anti-inflammatory experiment showed that the anti-inflammatory effects of 70％ ethanol extraction part ，ethyl acetate extraction part and water extraction part were the most significant （inhibitory rates of ear swelling were 54.07％，30.54％， 30.45％）. Five common peaks were determined in HPLC fingerprints of different polar parts from A. grossedentata . The spectrum-effect analysis results showed that the correlation of5 common peaks were higher than 0.6；among them ，peak 3 and peak 2 （dihydromyricetin） had the strongest anti- inflammatory effect ，and their correlation degrees were both mail：123745789@qq.com greater than 0.8. CONCLUSIONS ： The anti-inflammatory effect of A. grossedentata on xylene-induced ear swelling in mice is the result of multi-comp onent synergy ； unknown substance of peak 3 and dihydromyricetin may be the main active components of A. grossedentata .
ABSTRACT
OBJECTIVE：To establish a method for simultaneous determination of gallic acid ，protocatechuic acid ，mangiferin， homomangiferin，hyperoside and isoquercetin in Mangguo zhike tablets. METHODS ：HPLC was adopted. The determination was performed on Phenomenex Gemini C 18 column with mobile phase consisted of 0.1％ phosphoric acid water solution-acetonitrile （gradient elution ）at the flow rate of 1.0 mL/min. The detection wavelength was set at 258 nm. The column temperature was set at 30 ℃，and sample size was 5 μL. Gallic acid was used as the internal substance，and the correction factors of gallic acid ， protocatechuic acid ，mangiferin，homomangiferin，hyperoside and isoquercetin were calculated ；the results of quantitative analysis of multi-components by single marker method （QAMS） were compared with those of external standard method （ESM）. RESULTS：The linear range of gallic acid ，protocatechuic acid ，mangiferin，homomangiferin，hyperoside and isoquercetin were 45.75-183 0 μg/mL（r＝0.999 9），2.525-101.0 μg/mL（r＝0.999 9），65.33-261 3 μg/mL（r＝0.999 6），9.058-362.3 μg/mL（r＝ 0.999 9），3.885-155.4 μg/mL（r＝0.999 9），1.870-74.8 μg/mL（r＝0.999 9），respectively. The limits of quantification were 0.571， 0.643，1.053，0.854，0.830，1.500 μg/mL，respectively. The limits of detection were 0.171，0.193，0.316，0.256，0.249，0.450 μg/mL， respectively. RSDs of precision ，stability and reproducibility tests were all lower than 3％ . The average recoveries were 98.8％-101.9％（RSD＝1.3％ ，n＝6），94.3％-101.5％（RSD＝3.3％，n＝6），97.9％-100.5％（RSD＝0.9％，n＝6），98.2％- 101.6％ （RSD＝1.2％，n＝6），102.3％-106.1％（RSD＝1.3％，n＝6）， 96.6％ -99.3％（RSD＝1.0％ ，n＝6），respectively. RCFs of 73） protocate-chuic acid ， mangiferin， homomangiferin，hyperoside and isoquercetin were 0.568 3，0.500 3，0.687 6， 0.939 1 and 0.826 3，using gallic acid as internal substance . The content ranges of protocatechuic acid and other 4 com- ponents measured by QAMS were 0.197-0.440，3.262-11.250， 0.201-1.196，0.168-0.381，0.115-0.293 mg/tablet，respectively. The content ranges measured by ESM were 0.198-0.441，3.239-11.570，0.206-1.194，0.171-0.380，0.119-0.298 mg/tablet， respectively. By comparing the content determination results by QAMS and ESM ，the relative errors were －3.80％-0.74％，and there was no statistical significance （P＞0.05）. CONCLUSIONS ：The established QAMS method is accurate ，reliable and repeatable，and can be used for content determination of 6 medicinal components in Mangguo zhike tablets.