Analysis of Constituents of Seeds and Studies on the Comparison of Podophyllotoxin in Different Tissue from Podophyllum Hexandrum Royle
|School||Gansu Agricultural University|
|Course||Crop eco- physiological|
|Keywords||Sinopodophyllum emodi(Wall.)Ying Podophyllotoxin gas chromatography-mass spectrum Supercritical CO2 fluid extraction High performance liquid chromatography|
Podophyllotoxin(C22H22O8) belongs to Lignans class, mainly presence in the plant of Berberidaceae,Podophylloideae, Dysosma Woodson, Sinopodophyllum Ying, and Diphylleia Michx Podophyllum L, especially in the Sinopodophyllum emodi(Wall.)Ying has highest content of podophyllotoxin. Podophyllum plants are a class of substances containing significant biological activity, but also a long application history of medicinal plants. China’s Han Dynasty, "Shen Nong Ben Cao Jing" in there account of podophyllotoxin. Has been mainly used for snake bites, poison carbuncle swollen boils, bruises, rheumatism and bronchitis, sore muscles embolism; In abroad, American Sinopodophyllum emodi(Wall.)Ying (Podophyllum peltatum L.) is also used for medicinal. in recent years,as the pharmaceutical market increased demand for podophyllotoxin and the wild resources of Sinopodophyllum emodi(Wall.)Ying development and utilization, leading to lack of resources and Sinopodophyllum emodi(Wall.)Ying endangered.In 1992, it had been collected and listed by "Chinses Directory of Endangered Plant" and "Chinese Plant Red Date Book". In this study, gas chromatography-mass spectrometry (GC-MS) were used on analyzing the components of Sinopodophyllum emodi (Wall.) Ying seed and a certain foundation were lay on for seed development and utilization of resources; In addition, the use of methods of cell engineering technology for industrial production "podophyllotoxin "is being recognized as an effective way which protection and use of the scarce resources of Sinopodophyllum emodi (Wall.) Ying. Therefore, the comparison determination of the podophyllotoxin content in wild material and tissue culture materia have a great significance to screening the high "podophyllotoxin" training system in Sinopodophyllum emodi (Wall.) Ying tissue culture.In this experiment, wild Sinopodophyllum emodi (Wall.) Ying seeds and different in vitro tissue used as material then utilize GC-MS technique for analysis of seed composition; supercritical carbon dioxide extraction (CO2-SFE) technique were used to extracte,separate and comparative the podophyllotoxin in its different organs. The results obtained are as follows.1.GC-MS analysis of chem ical constituents of seeds from Sinopodophyllum emodi (Wall.) Ying. Use four different solvents such as ethyl acetate, ethanol, ethyl ether and chloroform toextract the chemical composition from seeds of Sinopodophyllum emodi (Wall.) Ying by soxhlet extraction and the products were analyzed by GC-MS.And then compare the effect of solvent extraction and indentify the organic compounds including, Provide the basis to the Sinopodophyllum emodi (Wall.) Ying resources for further development and utilization. The results show that the effect of different extraction solvents were as follows:ether> chloroform> anhydrous ethanol> acetate; The common substances which four types of extract all contains are: palmitic acid (Hexadecanoic acid), linoleic acid ((Z, Z)-9,12-Octadecadienoic acid), oleic acid, the content of linoleic acid was highest and the rest of the detected substances were mainly alkanes and aromatic compounds.2.The screening and optimization of process parameters for use of supercritical carbon dioxide (CO2-SFE) extraction of podophyllotoxin in Sinopodophyllum emodi (Wall.) Ying. Studied the several factors such asion pressure, extraction temperature, extraction time, flow rate of CO2,which affect supercritical carbon dioxide extraction (CO2-SFE) extract podophyllotoxin from Sinopodophyllum emodi (Wall.) Ying. according to Central Composite (Box-Benhnken) design principle of the response surface method (RSM) for optimization; using high performance liquid chromatography (HPLC) as a method for podophyllotoxin detection. The results showed that:The optimum conditions for use SFE-CO2 to extraction podophyllotoxin from Sinopodophyllum emodi (Wall.) Ying were:extraction pressure:38MPa, extraction temperature: 50℃, extraction time:52min, in these conditions, the extraction rate of podophyllotoxin up to 11.38%.3. Using the obtained optimum conditions on podophyllotoxin extraction in different organs from the wild and Tissue Sinopodophyllum emodi (Wall.) Ying, the results show that there was significantly different podophyllotoxin content in different materials, different tissues and organs, roots of wild:41.36μg/g; leaves:8.56μg/g; seeds:24.25μg/g; roots of tissue 60ds:6.17μg/g; leaves of tissue:0.84μg/g.