| 1.Objectives:To study the method of quality evaluation of Compound Flos Trollii Chinensis oral liquid and institute the quality criteria, based on researching the active compounds alignment, flavonoids.2.Methods:Research on the effective components, mainly flavonoids, in the oral liquid, and the method was as followed: Dried flowers were extracted with 70% ethanol. The concentrated extract was suspended in water and extracted successively with petroleum ether, ethyl acetate, and n-butanol. The n-butanol extract was isolated by column chromatography, such as silica gel, polyamide and Sephadex LH20. The structures of the components were elucidated by means of UV, NMR and other chromatographic technologies. The qualitation and quantitation are analyzed by TLC and HPLC separately. Data quality monitoring of technology: Test the preparation of oral liquid with the transferring rate of the main effective components as the index. Research on the quality evaluation of products: Take the flavonoids, the active compounds alignment, as the standards of chemical evaluation criterion. 3.Results:Seven compounds , mainly flavonoids, were isolated from the n-butanol extract after the 70% ethanol crude extract was extracted successively with petroleum ether, ethyl acetate, and n-butanol. Five compounds of it were determined by the analysis of physics, chemistry and spectroscopy, respectively 2"-O-β-L-galactopyranosyl orientin(I), orientin(II), vitexin(III), acacetin-7-O-neohesperidoside(IV), daucosterol(V) ,quercetin-3-O-sambubioside(VI) , quercetin-3-O-β-L-arabino (1′″-2")-β-D-glucoside) (VII)。The other two were still under determination. 2 "-O-β-L-galactopyranosyl orientin(I),acacetin-7-O-neohesperidoside(IV),quercetin-3-O-sambubioside(VI),quercetin-3-O-α-L-arabino(1′″-2")-β-D-glucoside) (VII) were obtained from Flos Trollii Chinensis for the first time. Results determined by HPLC: 2"-O-β-L-galactopyranosyl orientin had higher quantity than other flavonoids in the Flos Trollii Chinensis (The quantity of 2"-O-β-L-galactopyranosyl orientin of Aershan's Flos Trollii Chinensis reached 3.40%, which was the highest content among all the flavonoids in the Flos Trollii Chinensi from different places of production.), which possessed the specificity. Study and consummate the preparation technology of 2"-O-β-L-galactopyranosyl orientin. The method used for isolation and purification were convenient and economical, and the effect of isolation was favorable. The system of quality evaluation of Compound Flos Trollii Chinensis oral liquid was established by TLC and HPLC. TLC was used to establish the method of qualitation of index components of the preparation, 2"-O-β-L-galactopyranosyl orientin, orientin, vitexin, chlorogenic acid, baicalin. HPLC was used to establish the method of quantitation of index components of the preparation, 2"-O-β-L-galactopyranosyl orientin, orientin, chlorogenic acid, baicalin, and formulate the quanlity criteria of the quantitative determination of the index com component. The preparation had favorable stability and no evident toxic effects. The results were as followed: The regression equation of baicalin is Y=3829X-32.013,r=0.9998(n=5)(linear range 0.0968-0.3872μg),the content determined in the preparation was 15.37mg/ml,RSD=2.59% and the recovery rate was 99.66%,RSD=1.58%;The regression equation of chlorogenic acid was Y=2293.8X-14.913,r=0.9999(n=7)(linear range 0.0888-0.7104μg), the content of it determined in the preparation was 0.98mg/ml , RSD=2.89% and the recovery rate was 97.15%,RSD=1.47% ; The regression equation of orientin was Y = 2494X-6.2225,r=0.9999(n=5), (linear range 0.0392-0.1960μg),the content of it determined in the preparation was 1.86mg/ml,RSD=2.77% and the recovery rate was 102.04%,RSD=1.40%;The regression equation of 2"-O-β-L-galactopyranosyl orientin was Y=1343.5X-1.05 , r =0.9999(n=7)(linear range 0.0968-0.3872μg),the content of it determined in the preparation was 1.97mg/ml,RSD=1.72% and the recovery rate was 99.48%,RSD=1.79%。4.Conclusions : 2 "-O-β-L-galactopyranosyl orientin,acacetin-7-O-neohesperidoside,quercetin-3-O-sambubioside(VI),quercetin-3-O-α-L-arabino(1′″-2")-β-D-glucoside) (VII) were obtained from Flos Trollii Chinensis for the first time. The method(HPLC) of assaying of the 2"-O-β-L-galactopyranosyl orientin in Flos Trollii Chinensis and its preparation was established for the first time. The results showed that it had strong activity of antibiosis and antivirus, resembling that of some components such as orientin and vitexin, which ascertained the conclusion that flavonoids were the effective antibiosis and antivirus constituents of Flos Trollii Chinensis. Because 2"-O-β-L-galactopyranosy was the special component of the plants of the genus Trollius, we confirmed that it was the active compounds alignment of Flos Trollii chinensis and it could be one of the standards evaluating the quality of Flos Trollii chinensis and its preparation, which offered the scientific evidence to evaluate the quality of Flos Trollii Chinensis and its preparation in depth. |
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