The Cytotoxicity Of Stellera Chamaejasme L. Material Separation And Identification

Stellera chamaejasme L.,a member of the Stellerain the Thymelaeaeeae family, and widely distribute in north China, northwest, southwest and other places in China. In the local area of degraded grassland in north China it has become dominant population or to replace the original plants dominant position gradually. Stellera chamaejasme L.can greatly reduce the yield and quality of pasture, and become one of the important poison plants on grass which serious has influence the development of animal husbandry. Studies have shown that Stellera chamaejasme L. has antitumor, antiviral, antibacterial and insecticidal and other functions, and rich in resources, so the it has great value for research and development.Stellera chamaejasme L. has complicated composition and many active ingredients, but the main toxic substances and poisoning mechanism is not clear. Therefore the reasons greatly limit its further development and utilization. The study based on stellera chamaejasme L. materials focus on the toxic substances for extraction, separation, purification and structure identification. In order to make clear the cause of human and animal poisoning, and lay the foundation for its further development and use.In this article, we used water, ethanol and petroleum ether three reagents to extracte the root, stem leaf and flower of stellera chamaejasme L. respectively. We used9kinds crude extractings to determined cell toxicity by MTT experiments, and analyzed its cell inhibition rate. The results showed that: In48hours when reached the maximum inhibition rate, the petroleum ether extraction parts of root had the strongest toxicity. In0.2mg/ml, the cell inhibition rate for human embryonic kidney293A cells and liver cancer SMMC-7721reached46.978%and47.442%respectively. So we selected the petroleum ether extraction part of root for the further purification.Petroleum ether extraction of root eluted with petroleum ether, ethyl acetate proportion10:0to0:10on silica gel column chromatography for separation and purification. And test the components by thin-layer chromatography and high performance liquid chromatography (HPLC). Results showed that we can get some ingredients by the eluent petroleum ether, ethyl acetate proportion with8:2,6:4and5:5. Further study on three groups of elution components were determined by MTT cell toxicity test, results showed that:In48hours when reached the maximum inhibition rate, the eluent proportion part of6:4had the strongest toxicity. In0.2mg/ml, the cell inhibition rate for human embryonic kidney293A cells and liver cancer SMMC-7721reached38.850%and37.374%respectively.But the cell inhibition rate is lower than the petroleum ether extraction parts of root.We used elution components6:4for further purification and get white powder compound A and yellow oily mixture B. We determined the white powder compound A is a kind of terpenoids by MS and NMR datas. And its chemical structure is under testing.We used GC-MS to analyze the mixture B and found17substances.2,6,10,14,18,22Tetracosahexaene,2,6,10,15,19,23-h, gamma-sitosterol and (Z,Z,Z)-9,12,15-Octadecatrienal in stellera chamaejasme L. were found for the first time. And we have no idea about compound16. MTT cell toxicity experiment datas showed that mixture B has low cell toxicity, almost no inhibition to cells. But crystal A has high cell toxicity, and almost didn’t play a role of inhibition in48hours before, but in48-96hours, cell inhibitory activity and function is related to time and dose. In96hours, in0.2mg/ml, the cell inhibition rate for human embryonic kidney293A cells and liver cancer SMMC-7721reached24.259%and56.694%respectively. By datas We can preliminary judge that the compound A is the main cell toxic substances of stellera chamaejasme L.