Study On The Compounds Of Castanea Mollissima Blume

Castanea mollissima Blume Shell is involucre, sexual sweet, astringent taste, non-toxic, tonifying the kidney and strengthening the spleen and replenishing qi and reinforcing ribs, dispersion effect can be treated with detoxification, swelling, erysipelas, scrofula diseases. Castanea mollissima Blume is Fagaceae Castanea plants. The roots, bark, leaves, flowers, exocarp, endocarp, involucre of Castanea mollissima Blume can be used as medicine. The main topic of this paper is studying on the chemical composition of Castanea mollissima Blume Shell, and to provide a basis for further development and utilization of Castanea mollissima Blume Shell.This paper is mainly divided into three parts, the first part for extraction of chemical composition from Castanea mollissima Blume Shell, the second part of the the isolated compounds were screened, the establishment of the third part is the characteristics of Castanea mollissima Blume Shell.The dry powder of Castanea mollissima Blume Shell was extracted with 95% ethanol. Portions of petroleum ether, ethyl acetate, n-butanol and water were obtained by extraction. In this paper, the part of ethyl acetate was isolated and purified. Seven compounds were isolated by the method of ordinary pressure normal phase silica gel column chromatography, reversed phase silica gel column chromatography, Sephadex LH-20, recrystallization etc. Three compounds were identified through compared with the control sample. They are β-Sitosterol(2)、Olean acid(3)、kaempferol(6).The structures of four compounds were elucidated by means of TLC,1H-NMR,13C-NMR. Detailed Results As Follows, Toluene 2,6-2 methyl carbamate(1)、(3-Sitosterol(2)、Olean acid(3)、Apigenin (4)、Betulinic acid (5)、kaempferol(6)、gallic acid(7). Compound 4 and Compound 5 was isolated from this plants for the first time.The use of human breast cancer cells MCF-7、neural tumor cells PC-12、liver cancer cells SMMC-7721 to study on the anticancer activity of compound 4、5、6 by the method of CCK8.The experimental results showed that three compounds had no significant inhibitory effect on the PC-12, but for MCF-7、 PC-12 had certain inhibitory effect.Using HPLC method, determine kaempferol, quercetin, gallic acid, ursolic acid of Castanea mollissima Blume Shell. The chromatographic analysis of Castanea mollissima Blume Shell was carried out on a InertSustain C18(5um,4.6×250mm) column with acetonitrile-0.4% phosphoric acid (0-20min,4%～5%; 20～21min,5%～28%; 21-70min,28%～33%; 70～71min,33%～93%; 71～90min,93%-95%) as the mobile phase. The detection wavelength of gallic acid was set at 270nm. The detection wavelength of kaempferol and quercetin was set at 365nm. The detection wavelength of ursolic acid was set at 210nm. The column temperature was 30℃, and the flow rate was about 0.8 mL/min. The regression equation of gallic acid was y=3353.8x+58157(r=0.9997). The linear was 36-360μg. The regression equation of kaempferol was y=5680.3x+3969.3 (r=0.9997). The linear was 36-360μg. The regression equation of quercetin was y=3751.4x+574.79 (r=0.9996). The linear was 9-90μg. The regression equation of ursolic acid was y=575.55x-2305.8 (r=0.9997). The linear was 71.6-716μg.