Study On The Chemical Constituents Of The Alkaloids From The Root Of Aconitum Flavum Hand.-mazz And Its Anti-tumor Activities

Objective：Enriched alkaloid fraction and non-alkaloid fraction were obtained fromthe ethanol extract of Aconitum flavum Hand.-Mazz. by cation exchange resins. Thenthe ethanol extract, alkaloid fraction and non-alkaloid fraction were tested for antitumor activities by MTT. Six compounds were isolated from the alkaloid fraction. MTT assay was used to screen chemical compounds of the active ingredient of Aconitum flavum Hand.-Mazz.. This study will provide the basis for the exploition and utilizationof Aconitum flavum Hand.-Mazz.Methods:1. Microwave-assisted reflux extraction was used to extract the dry roots of Aconitum flavum Hand.-Mazz.2times diluted the extractum with20%alcohol and acidified with hydrochloric acid to PH=1-2. Then alkaloid fraction and non-alkaloid fractionwould be enriched by the technology of macroporous adsorptive resins2. Using MTT assay to determine the growth inhibition effect of different extracts of tiebangchui to human gastric carcinoma SGC-7901cells, hepatic carcinoma HepG2cells and lung cancer A549cells, aimed at choosing the most effective extract partto do further reaearch.3. The antitumor active site was separated by silica gel column chromatography,ODS column chromatography and gel column chromatography and thin-layer chromatography and other separation methods for monomer compounds, In combination with1H-NMR,13C-NMR spectrum confirmed the structure of those monomer compounds. 4. MTT assay was been used for screening those6compounds in the same waywhich was seperated from the active parts on SGC-7901, HepG2and A549to getMonomer compounds with antitumor activity.Results:1. Crude extract0.8kg was extracted from the Herbs10kg, the macroporous resin parts was enriched to79.3g by30%alcohol;30%alcohol active extract parts70g was separated by a cation exchange resin.Then we obtained the non-alkaloid fraction33g, and the alkaloid fraction30.5g2. The antitumor activities of the ethanol extract, alkaloid fraction and non-alkaloid fraction from Aconitum flavum Hand.-Mazz. were measured by MTT assay. The results show that SGC-7901’s IC50were125.83μg/ml,27.92μg/ml and421.42μg/ml, respectively; HepG2’s IC50were26.18μg/ml,15.22μg/ml and745.91μg/ml, respectively; A549’s IC50were386.3μg/ml,37.16μg/ml and433.24μg/ml, respectively.3. Six compounds were isolated and identified from active parts, including4-(2-formyl-5-hydroxy methylpyrrol-1-yl) butyric acid (Ⅰ), neoline(Ⅱ),14-O-acetylneoline(Ⅲ),songorine(Ⅳ),12-epi-napelline(Ⅴ)and12-epi-dehydronapelline(Ⅵ).4. The antitumor activity of6compounds isolated from Aconitum flavum Hand.-Mazz. were measured by MTT assay. The results show that SGC-7901’s IC50were16.43μg/ml,8.13μg/ml,16.64μg/ml,23.26μg/ml and23.24μg/ml, respectively; HepG2’s IC50were12.41μg/ml,16.17μg/ml,31.36μg/ml,34.82μg/ml and16.67μg/ml, respectively;A549’s IC50were15.20μg/ml,8.98μg/ml,22.13μg/ml,23.66μg/ml and27.12μg/ml, respectively.Conclusion:1. The alkaloid fraction is the principal anticancer extract from Aconitum flavumHand.-Mazz. 2.6compounds were isolated and identified from active parts. One was4-(2-formyl-5-hydroxy methylpyrrol-1-yl) butyric acid(Ⅰ);neoline(Ⅱ);14-O-acetylneoline(Ⅲ); songorine(Ⅳ);12-epi-napelline(Ⅴ) and12-epi-dehydronapelline(Ⅵ).4-(2-formyl-5-hydroxymethylpyrrol-1-yl) butyric acid (Ⅰ) and14-O-acetylneoline(Ⅲ) were first isolated from Aconitum flavum Hand.-Mazz.3. neoline(Ⅱ);14-O-acetylneoline(Ⅲ); songorine(Ⅳ);12-epi-napelline(Ⅴ) and12-epi-dehydronapelline(Ⅵ) could inhibit the growth of SGC7901, HepG2and A549line.