Study On Quality Control Of Solanum Lyratum Thunb

The present paper developed the methods to evaluate the quality of Solarium lyratum Thunb. systematiclly. The concentration of general saponins, general flavonoids, general alkaloids, diosgenin, and six phenolic compounds in Solarium lyratum Thunb. were determined, respectively. The HPLC fingerprint analysis for the quality control of Solarium lyratum Thunb. was established. Moreover, the antitumor activity in vitro was investigated for extract of Solarium lyratum Thunb..Firstly, the orthogonal test was used to confirm the optimal extraction conditions of general saponins in Solarium lyratum Thunb. by investigating concentration of hydrochloric acid (mol·L^-1), volume fraction of ethanol（%）, and time of hydrolysis. The assay of general saponins was developed with colorimetry of colorating with perchloric acid,as diosgenin control sample.The contents of general saponins in 19 batches of Solarium lyratum Thunb. have determined by UV-Vis. The linear range of general saponins between 8.91 to 29.70μg·mL^-1 and the average recoveries was 95.9% （RSD = 1.5%）.Secondly, the orthogonal test was used to confirm the optimal extraction conditions of general flavonoids in Solarium lyratum Thunb. by investigating volume portions of ethanol （%）, extraction numbers and extraction time. The content of general flavonoids was determined with colorimetry of colorating with AlCl₃, rutin as control sample. The contents of general flavonoids in 21 batches of Solarium lyratum Thunb. have determined by UV-Vis. The linear range of general flavonoids between 5.48 to 21.92 ug·mL^-1 and the average recoveries was 96.7% （RSD = 1.1%）.Thirdly, the orthogonal test was used to confirm the optimal extraction conditions of general alkaloids in Solarium lyratum Thunb. by investigating volume portions of ethanol （%）, extraction numbers and volume of ethanol. The content of general alkaloids was measured by acid dye colorimetry at 600nm, strychnine as control sample, and bromothymol blue (0.001mol·L^-1) as the acid dye.The contents of general alkaloids in 21 batches of Solarium lyratum Thunb. have determined by UV-Vis. The linear range of general alkaloids between 10.20 to 35.70μg·mL^-1, and the average recoveries was 99.3% （RSD = 1.3%）.Fourthly, the orthogonal test was used to confirm the optimal extraction conditions of diosgenin in Solarium lyratum Thunb. by investigating volume of ethanol, concentration of sulfuric acid and hydrolysis time. The content of diosgenin in 20 batches of Solarium lyratum Thunb. by RP-HPLC, using Hypersil ODS₂ as column （4.6mm×200mm,5μm）, mixture of acetonitrile and water （80:20, V/V） as mobile phase, and the detection wavelength at 210nm. The linear range of diosgenin between 0.20 to 4.02μg·mL^-1, and the average recoveries was 96.7% （RSD = 1.6%）.Fifthly, the simultaneous assay of six phenolic compounds in Solarium lyratum Thunb. was developed by HPLC. Diamonsil C18 column （4.6mm×250mm, 5μm） was used, with formic acid solution （0.3%） and mixture of methanol and acetonitrile （4:1, V/V） as mobile phase in a gradient mode, and the detection wavelength at 254nm. The orthogonal test was again used to confirm the optimal extraction conditions of phenolic compounds by investigating volume portions of ethanol （%）, extraction numbers and volume of ethanol. The linear ranges of protocatechuic acid, chlorogenic acid, vanillic acid, caffeic acid, ferullic acid, and rutin were 0.14-6.30μg·mL^-1, 8.80-396μg·mL^-1,0.78-35.28μg·mL^-1,0.42-19.08μg·mL^-1, 0.43-19.44μg·mL^-1, and 3.20-144μg·mL^-1, respectively. And the average recoveries of six phenolic compounds were 98.2% （RSD=1.4%）, 100.1% （RSD=1.5%）, 97.8% （RSD=1.1%）, 98.2% （RSD=0.6%）,99.7% （RSD=0.8%）, and 99.1% （RSD=0.6%） respectively.Sixthly, the RP-HPLC fingerprint analysis for the quality control of Solarium lyratum Thunb. was established. Kromasil C18 column （250mm×4.6mm, 5μm） was used, with mixture of acetonitrile and formic acid solution （0.3%） as mobile phase in a gradient mode, the flow rate was 1.0 mL·min^-1, and the detection wavelength at 270nm. 21 batches of Solarium lyratum. Thunb. were detected under the conditions above. The hierarchical cluster analysis by SPSS 13.0 was applied to the fingerprints of Solarium lyratum Thunb. and the 21 batches of sample were classified into 4 grades. The mutual model from 12 batches of sample were established by the results of cluster analysis and concering the main producing area of herbs. Furthermore, the quality of samples was assessed by Similarity Evaluation System for Chromatographic Fingerprint of TCM 2004.Finally, the antitumor activity of ethanol extract from 18 batches of Solarium lyratum Thunb. was studied by MTT method. Human cervical carcinoma Hela cells cultivated in vitro was selected as model and IC50 was calculated. The results showed the differences of proliferation inhibiting effect and yield of extract occurred among Solarium lyratum Thunb. from different origin, and proliferation inhibiting effect in vitro didn’t relate well to yield of extract.