Study On The Chemical Constituents Of Artificially Cultivated Antrodia Camphorata Fruit Bodies

Antrodia cinnamomea is a precious endemic Taiwanese medicinal fungus.In recent years,it has received more and more attention due to its anti-tumor,anti-viral,anti-oxidation,anti-inflammatory and hypoglycemic effects.In this study,the artificial culture of A.cinnamomea fruit body was selected as the research object.The silica gel column chromatography method was used to separate and purify the fruit body of A.cinnamomea,and 19 compounds were obtained.15 compounds were identified by 1H-NMR,13C-NMR and MS spectroscopy,respectively:quinone Q3(1),1,3-dimethoxybenzoic acid(2),2,3,6-trimethoxy-5-methylphenol(3),vanillin(4),dibutyl phthalate(5),gallic acid(6),2,3-dimethoxy-1,4-benzenediol(7),?-carotene(8),antrodin A(9),ursolic acid(10),himanimides A(11),D-sorbitol(12),uracil(13),(5S-5-hydroxy-3-p-L-ribofuranosyl-2(5H)-furanone(14),Adenosine(15).Among them,compounds 5,7,and 11-15 were isolated from A.cinnamomea for the first time,and compounds 1,3,and 9 were isolated from the fruit body of A.cinnamomea for the first time.In this study,the response surface method was used to optimize the ultrasonic-assisted extraction process of the triterpenoids of A.cinnamomea,and the optimal extraction conditions were determined by single factor experiment:ethanol concentration was 85%,ratio of material to liquid was 1:20,ultrasonic time was 80 min.Finally,it was verified that the total yield of A.cinnamomea was 2.1306%.The antibacterial activity of the extract of A.cinnamomea and its different extracts was studied.The results showed that the extract and water fraction of A.cinnamomea had the strongest antibacterial activity against Escherichia coli and the weakest antibacterial effect against Bacillus subtilis;water fraction had the strongest antibacterial activity and the weakest antibacterial effect against Staphylococcus aureus.The minimum inhibitory concentration(MIC)of three parts on Escherichia coli,Bacillus subtilis and Staphylococcus aureus were as follows:A.cinnamomea extract:2 mg·mL-1;Petroleum ether extract layer:2 mg·mL-1,2 mg·mL-1 and 4 mg·mL-1;aqueous phase:4 mg·mL-1,8 mg·mL-1 and 8 mg mL-1.Based on the above separation,the three monomeric compounds quinone Q3,antrodin A and himanimides A were quantitatively analyzed and tested by high performance liquid chromatography(HPLC).Chromatographic conditions:column:WondaCract ODS-2 C18;mobile phase:acetonitrile-5%aqueous methanol(75:25);flow rate:1 mL·min-1,column temperature:30 ?;injection volume:20 ?L;detection wavelength:205 nm.The methodological verification results show that the precision of the instrument is good;the method has high reproducibility;the stability of artificially cultured A.cinnamomea fruit samples is good.The results of the content determination showed that the contents of antrodin A?himanimides A and quinone Q3 in artificial cultured Antrodia camphorata were 0.3549 mg·g-1,0.1801 mg·g-1 and 1.1003 mg·g-1,,respectively.