木瓜超氧化物歧化酶、黄酮及齐墩果酸的综合提取初探
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摘要
木瓜(Fructus Chaenomelis)是临床常用中草药之一,药理实验研究表明,木瓜具有保肝、抗菌等作用。木瓜中含有黄酮、齐墩果酸等多种有效药用成分,还被认为是良好的植物超氧化物歧化酶(SOD)来源,本文对其综合利用进行了研究。
由于目前尚未有文献对木瓜中SOD含量测定或提取的报道,本文首先对新鲜木瓜进行SOD的提取和活力测定,并与其他几种常见蔬菜包括青椒、豌豆、大蒜黄豆进行比较。采用邻苯三酚自氧化法和氮蓝四唑(NBT)光还原法分别对它们进行活力的测定,讨论了两种测定方法的优缺点,最后确定NBT光还原法更适合本课题中酶活力的测定,根据两种不同方法对SOD酶活力测定结果进行换算,确定了两种方法测定结果之间的转换系数为3.5～3.98,并测定出木瓜中SOD的活力为55.9U/g(FW)·h。低于其他被测的几种常见蔬菜。根据上述结果,指出木瓜不是工业提取植物SOD的合适原料。
其次,用提取SOD后晾干的木瓜渣联合提取黄酮和齐墩果酸。提出了综合提取工艺流程:木瓜以95%乙醇渗漉,渗漉液浓缩、水洗、多次酸碱沉淀,获得齐墩果酸,采用紫外分光光度法测得其得率为0.0146%;渗漉后残渣用70%乙醇回流提取,聚酰胺柱层析分离,获得黄酮,采用硝酸铝络合分光光度法测得其得率为0.307%。上述结果表明黄酮提取工艺基本可行,齐墩果酸提取工艺有待改进。
Papaya (Fructus Chaenomelis) is one of the commonly used traditional Chinese herbal medicines. Pharmacological studies indicate that papayas have liver protection and antibacterial effects. Papaya contains flavonoids, oleanolic acid and other active ingredients, it is also considered to be a good source of botanic superoxide dismutase (SOD). In this thesis, the comprehensive utilization of papayas was studied.
There is no report about the method for determination or extraction of SOD from papaya, in this paper, first, SOD was extracted from fresh papayas and the activity of SOD was determined, the result was compared with those of several other common vegetables, including green peppers, peas, garlic and soybeans. The activity of SOD was determined by using pyrogallol autoxidation method and NBT-reduction method, respectively, and found that NBT-reduction method is more suitable for SOD detection in the extraction of papayas. According to the results of two different methods, the transfer coefficient of the results of two SOD detection methods was found to be 3.50～3.98. It was found that the activity of SOD in papaya is only 55.9 U/g (FW) ?h, which is lower than that of above mentioned plants. This result means that papaya is not a suitable raw material for industrial extraction of botanic SOD.
Second, dried residue of the papaya after SOD extraction is used for extracting flavonoids and oleanolic acid. The comprehensive extraction method was suggested. Dry papaya was percolated with 95% ethanol and percolate was condensed, washed with water, repeatedly precipitated with acid and base and then oleanolic acid was obtained. The content of oleanolic acid was determined by UV spectrophotometry and found the yield of oleanolic acid was 0.0146%. Residue after percolated was extracted with 70% ethanol and segregated with polyamide column chromatography, and then flavonoids was obtained , the content of total flavonoids was determined using aluminum nitrate complex spectrophotometry method and found the yield of total flavonoids was 0.307%.These results indicate flavonoids extraction process is feasible, however, the oleanolic acid extraction process should be improved.
由于目前尚未有文献对木瓜中SOD含量测定或提取的报道,本文首先对新鲜木瓜进行SOD的提取和活力测定,并与其他几种常见蔬菜包括青椒、豌豆、大蒜黄豆进行比较。采用邻苯三酚自氧化法和氮蓝四唑(NBT)光还原法分别对它们进行活力的测定,讨论了两种测定方法的优缺点,最后确定NBT光还原法更适合本课题中酶活力的测定,根据两种不同方法对SOD酶活力测定结果进行换算,确定了两种方法测定结果之间的转换系数为3.5～3.98,并测定出木瓜中SOD的活力为55.9U/g(FW)·h。低于其他被测的几种常见蔬菜。根据上述结果,指出木瓜不是工业提取植物SOD的合适原料。
其次,用提取SOD后晾干的木瓜渣联合提取黄酮和齐墩果酸。提出了综合提取工艺流程:木瓜以95%乙醇渗漉,渗漉液浓缩、水洗、多次酸碱沉淀,获得齐墩果酸,采用紫外分光光度法测得其得率为0.0146%;渗漉后残渣用70%乙醇回流提取,聚酰胺柱层析分离,获得黄酮,采用硝酸铝络合分光光度法测得其得率为0.307%。上述结果表明黄酮提取工艺基本可行,齐墩果酸提取工艺有待改进。
Papaya (Fructus Chaenomelis) is one of the commonly used traditional Chinese herbal medicines. Pharmacological studies indicate that papayas have liver protection and antibacterial effects. Papaya contains flavonoids, oleanolic acid and other active ingredients, it is also considered to be a good source of botanic superoxide dismutase (SOD). In this thesis, the comprehensive utilization of papayas was studied.
There is no report about the method for determination or extraction of SOD from papaya, in this paper, first, SOD was extracted from fresh papayas and the activity of SOD was determined, the result was compared with those of several other common vegetables, including green peppers, peas, garlic and soybeans. The activity of SOD was determined by using pyrogallol autoxidation method and NBT-reduction method, respectively, and found that NBT-reduction method is more suitable for SOD detection in the extraction of papayas. According to the results of two different methods, the transfer coefficient of the results of two SOD detection methods was found to be 3.50～3.98. It was found that the activity of SOD in papaya is only 55.9 U/g (FW) ?h, which is lower than that of above mentioned plants. This result means that papaya is not a suitable raw material for industrial extraction of botanic SOD.
Second, dried residue of the papaya after SOD extraction is used for extracting flavonoids and oleanolic acid. The comprehensive extraction method was suggested. Dry papaya was percolated with 95% ethanol and percolate was condensed, washed with water, repeatedly precipitated with acid and base and then oleanolic acid was obtained. The content of oleanolic acid was determined by UV spectrophotometry and found the yield of oleanolic acid was 0.0146%. Residue after percolated was extracted with 70% ethanol and segregated with polyamide column chromatography, and then flavonoids was obtained , the content of total flavonoids was determined using aluminum nitrate complex spectrophotometry method and found the yield of total flavonoids was 0.307%.These results indicate flavonoids extraction process is feasible, however, the oleanolic acid extraction process should be improved.
引文
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