| This paper is composed of three chapters, the first chapter introduced the component of endophytic fungus secondary metabolites from Paris; the second chapter showed the specific process of extraction and separation, screening and identification the activity of endophytic fungus secondary metabolites in Paris; the third chapter is the determination of several butyrolactone compounds from the fermentation products of an endogenetic fungi aspergillus versicolor with the help of high performance liquid chromatography.The chemical constituents of Phomopsis genus are complex, they have better pharmacological activities and have a relatively wide use in clinical application. Paris belong to Phomopsis genus, also known as Paris root, Tgiganteum, Yunnan Paris and so on, can be heat-clearing, antibiotic inhibition, sedation and analgesia, alleviate the pain and swelling, stop the bleeding, immunoregulation, antitumor, anti-inflammatory and the like.The secondary metabolites of endophytic fungus are important resources to found new compounds possess antibacterial activity, antitumor activity and anti HIV activity. It has good prospect for development that isolated active ingredients from the universality and diversity of endophytic fungus and secondary metabolites which is a new way to solve the shortage of medical resources. In this paper, by means of TLC, column chromatography method to separation and purification the secondary metabolites separated from the endophytic fungi aspergillus versicolor. The structure of secondary metabolites which have been identified by mass spectrometry, nuclear magnetic resonance, polarimeter, ultraviolet, infrared. Finally obtained 20 butyrolactone compounds including 20 new, they are terrephenol A(1*), terrephenol B(2*), asperphenol A(3*), asperphenol B(4*), versicolactone A(5*), versicolactone B(6*), versicolactone C(7*), versicolactone D(8*), aspernolide C(9*), aspernolide D(10*), aspernolide A(11), butyrolactone III(12), butyrolactone I(13), rubrolide S(14), aspernolide E(15), 4-(4-hydroxyphenyl)-5-(4–hydroxyphenylmethyl)-2-hydr oxyfurane-2-one(16), aspernolide B(17), butylrolactone-VI(18), butyrolactone V(19), butylrolactone IV(20).We screened the inhibitory activities against TMV virus of the compounds 1*-20 with ningnamycin as controls. Results showed that compounds 1*, 2*, 4*, 8*, 20 have better anti TMV activity, inhibition rates were 35.2%, 31%, 46.7%, 46.4%, 35.4%, compounds 3*, 5*, 6*, 9*, 11, 12, 13, 15, 16, 17, 18, 19 provide excellent resistance to tobacco mosaic virus activity, inhibition rates were 22.5%, 22.6%, 25.4%, 22.5%, 24.6%, 20.8%, 21.8%, 25.2%, 21.8%, 28.4%, 21.5%, 23.5%, compounds 7*, 10*, 14 with moderate anti TMV activity, inhibition rates were 18.6%, 18.6, 15.4%.Also did the screening of cytotoxic activity to the new compounds 5*, 6*, 7*, 8*, the results showed that compound 8* has better function against A549 and MCF7, the IC50 values were 3.2, 2.5 μM.This article uses the high-speed homogenate method to extract the chemical composition. The extration was skimed by the small column of solid phase extraction with MCI-GEL reverse phase resin, using the HPLC to determined the 6 components(aspernolide E, butyrolactone II, butyrolactone IV, butyrolactone III, butyrolactone I, aspernolide A) under the condition of wavelength with 295 nm. Results showed the detection limit is ≦ 55 g/L, linear range between 0.30-280 g/m L, recovery rate is 95.8%-105.9%, RSD value is 1.80%-2.96%, which suggested that the method is accurate. Accurate determination of the content is an important guidance for the selection and control of fermentation conditions of biology. |
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