Antitumor Activity Of Frankincense,Myrrh And Compatible Drugs Based On CO2 Supercritical Extraction On Volatile Components

Objective:1.To investigate the effect of Frankincense,Myrrh and Myrrh on malignant tumor cell lines（Breast cancer cell line MCF-7、Human liver cancer cell line Bel-7402,human lung cancer cell line A549）Effects of in vitro proliferation.2.Further explore the effects of different solvents for Frankincense,Myrrh and Myrrh extracted on the volatile components on the proliferation of malignant tumor cell lines in vitro.3.Determination of Antitumor Active Components of Frankincense and Myrrh by GC-MS.Methods:1.Supercritical CO₂extraction process for volatile oil of Frankincense and Myrrh was optimized by apply orthogonal test.2.Isolate and purify the constituents of Frankincense Myrrh essential oil by systematic solvent separation.3.Comparison of Frankincense,Myrrh,and Myrrh on tumor inhibition in vitro:Mix different concentrations of Frankincense,Myrrh,and Myrrh to the extract.Directly added to a 96-well culture plate containing tumor cells and cultured for 24hours,the MTT method was used to determine its effect on three human malignant tumor cell lines（human breast cancer cell line MCF-7,human liver cancer cell line Bel-7402,and human lung cancer Effect of cell line A549）colonization.4.Comparison of antitumor activity of Frankincense and Myrrh components extracted with different solvents in vitro:Different concentrations of petroleum ether,ethyl acetate,n-butanol,water extract,after 24 hours in a 96-well culture plate containing tumor cells,the effects of MTT on the proliferation of three human malignant tumor cell lines（human breast cancer cell line MCF-7,human liver cancer cell line bel-7402,and human lung cancer cell line A549）were deter mined.Results:1.The optimal process for extracting volatile components of Frankincense and Myrrh by supercritical CO₂extraction is The extraction pressure was 30 MPa,the extraction time was 2 h,the extraction temperature was 55℃（separation temperature1 was 55℃,separation temperature 2 was 40℃）,and the extraction yield was the highest when the entrainer anhydrous ethanol dosage was 150 m L.2.Comparison of antitumor activity of Frankincense,Myrrh,and Frankincense Myrrh on volatile oil components:（1）The extracts of the three medicinal herbs showed significant inhibitory effects on tumor cells in three different tissues of breast cancer MCF-7 cell line,liver cancer Bel-7402 cell line,and lung cancer A549 cell line after24 h,Its inhibition rate:volatile oil of Myrrh>milk Myrrh pair>Frankincense.And the inhibitory effect increases with the increase of drug concentration,showing a dose-dependent relationship.（2）When the drug concentration is above 50μg·ml^-1,the tumor cell inhibition rate reaches 10%.When the drug concentration is 500μg·ml^-1,the tumor cell inhibition rate reaches 50%.3.Comparison of tumor inhibition of Frankincense,Myrrh and different solvent extracts:Oil ether,ethyl acetate and n-butanol were used to extract Frankincense,Myrrh and its volatile oil,found that the four components had a certain inhibitory effect on the proliferation of the three types of tumor cells,and the inhibition rate of the aqueous component of ethyl acetate extract>n-butanol>petroleum ether>was reduced.The inhibition was dose dependent.4.Combined with the screening results of anti-tumor activity,GC-MS was used to analyze the anti-tumor components of component 2,it was found that the content of eucalyptol was the highest,γ-cedrene,octyl acetate,（8R,8AS）-8,8a-dimethyl-2-（propane-2-alkylene）-1,2,3,7,8,8a-Hexahydronaphthalene andγ-elemene followed.Conclusions:1.The extraction rate of volatile oil of Frankincense and Myrrh by supercritical CO₂extraction method is significantly higher than that of steam distillation,and there are certain differences in the composition.2.Frankincense,Myrrh,and Myrrh have certain inhibitory effects on the proliferation of the selected three tumor cells,of which the Myrrh has the strongest inhibitory effect,and the same dose of compatible drugs has not shown a synergistic effect.3.GC-MS analysis found that the anticancer active component of Myrrh had the highest content of cineole,γ-cedrene,octyl acetate,（8R,8AS）-8,8a-dimethyl-2-（propane-2-Alkylene）-1,2,3,7,8,8a-hexahydronaphthalene,followed byγ-elemene.