【作者】 周颖；

【导师】 李俊丽；

【作者基本信息】 武汉理工大学 ， 药学， 2023， 硕士

【摘要】 甜瓜,是葫芦科黄瓜属蔓性草本植物,在我国的栽培与食用历史悠久。甜瓜植株兼具食用价值与药用价值,其中富含黄酮、皂苷化合物,但目前对于甜瓜中黄酮、皂苷化合物的提取分离纯化方法不够完善,且对分离纯化得到的化合物的药用活性研究报道较少。本研究选取甜瓜为实验对象,系统地研究了甜瓜药用部位中黄酮、皂苷化合物的提取、纯化工艺以及纯化物的药用活性。(1)选取甜瓜茎、甜瓜叶、甜瓜子作为实验对象,通过单因素实验系统地考察了液料比、提取时间、提取温度及提取剂浓度等因素对黄酮、皂苷水提法与醇提法提取量及提取率的影响。在此基础上,采用正交试验设计法优选了各部位黄酮、皂苷水提法与醇提法的最佳提取条件。其中甜瓜子的水提法最佳工艺为:液料比20∶1、提取温度70℃和提取时间7 h;甜瓜子的醇提法最佳工艺为:液料比20∶1、提取温度70℃、提取时间6 h和乙醇浓度70%。(2)通过比较三种树脂(D-101、AB-8、S-8)对甜瓜子黄酮、皂苷的静态吸附率与解吸率,发现D-101大孔吸附树脂性能最佳。通过考察各因素对D-101大孔吸附树脂纯化甜瓜子黄酮、皂苷的影响,得出最优纯化方案。甜瓜子样品溶液以0.89 mL/min的吸附流速通过D-101大孔吸附树脂。吸附后经2 BV的去离子水以0.67 mL/min的流速去除杂质,使用60%乙醇以1 mL/min流速进行洗脱,甜瓜子黄酮与皂苷的洗脱效果较好。(3)对甜瓜子20%、40%、60%乙醇洗脱组分进行体外抗氧化活性测定,结果表明甜瓜子60%乙醇洗脱组分的羟基自由基清除率及总还原力较高,抗氧化能力较好。将甜瓜子60%乙醇洗脱组分按收集管数分为样品A、B、C,研究其在不同浓度下对大肠杆菌及金黄色葡萄球菌生长的抑制效果。结果表明,样品A、B、C对金黄色葡萄球菌抑制效果较好,其中样品B对金黄色葡萄球菌的抑制效果好于样品A与样品C,在5 mg/m L及10 mg/m L的药物浓度下对金黄色葡萄球菌的抑制率分别为97.39%及99.9%。探究其在不同浓度下对小鼠胶质瘤细胞活性的影响。其中样品B对小鼠胶质瘤细胞的生长的抑制效果最好,当药物浓度为1 mg/m L时,样品B的添加可以使小鼠胶质瘤细胞活性下降至原本的14.50%。综上,本研究系统地研究甜瓜茎、甜瓜叶、甜瓜子中黄酮、皂苷化合物的提取分离纯化方法,形成高效、高得率、高回收率的黄酮、皂苷化合物的提取工艺,确定合适的纯化工艺,为甜瓜植物黄酮、皂苷的提取分离提供较为详尽的技术方案与基础数据。考察了甜瓜子纯化物的体外药用活性,为合理开发利用甜瓜天然活性成分资源提供研究基础与科学依据。更多还原

【Abstract】 Muskmelon,which is a kind of herb of cucumber in calabash family,has a long history in cultivation and edible in our country.Muskmelon plants have both edible value and medicinal value,which are rich in flavonoids and saponins.However,the extraction,separation and purification methods of flavonoids and saponins in muskmelon are not perfect,and there are few reports on the medicinal activity of the separated and purified compounds.However,the extraction,separation and purification methods of flavonoids and saponins in muskmelon are not perfect,and there are few reports on the medicinal activity of the separated and purified compounds.In this study,muskmelon was selected as the experimental object,and the extraction and purification process of flavonoids and saponins in the medicinal parts of muskmelon and the medicinal activity of the purified products were systematically studied.(1)The medicinal parts such as muskmelon stems,leaves and seeds were selected as experimental objects,and the flavonoids and saponins were extracted by water extraction and alcohol extraction.The effects of liquid-solid ratio,extraction time,extraction temperature and extractant concentration on the extraction amount and extraction rate of flavonoids and saponins were systematically investigated by single factor experiment.On this basis,the optimum extraction conditions of water extraction and alcohol extraction of flavonoids and saponins in each part were optimized by orthogonal test design.For muskmelon seeds,the optimum conditions of water extraction were as follows: liquid-solid ratio of 20:1,extraction temperature of 70°C and extraction time of 7 h;the optimum extraction conditions of alcohol extraction were as follows: liquid-solid ratio of 20:1,extraction temperature of 70°C,extraction time of 6 h and ethanol concentration of 70%.(2)By comparing the static adsorption rate and desorption rate of three resins(D-101,AB-8,S-8)on flavonoids and saponins in muskmelon seeds,it was found that D-101 macroporous adsorption resin had the best performance.By investigating the effects of various factors on the purification of flavonoids and saponins from muskmelon seeds by D-101 macroporous adsorption resin,the optimal purification scheme was obtained.The muskmelon seed sample solution passed through the D-101 macroporous adsorption resin at an adsorption flow rate of 0.89 mL/min.After adsorption,the removal of impurities was removed by twice the volume of deionized water at a flow rate of 0.67 mL/min,and 60% ethanol was eluted at a flow rate of 1mL/min.(3)The results showed that the hydroxyl radical scavenging rate and total reducing power of 60% ethanol elution fraction were significantly higher than those of 20% ethanol elution fraction and 40% ethanol elution fraction.Divide the 60%ethanol elution component of muskmelon seeds into samples A,B and C according to the number of collecting tubes,and study its inhibition effect on the growth of Escherichia coli and Staphylococcus aureus at different concentrations.The results showed that sample A,B and C had a good inhibitory effect on Staphylococcus aureus,and sample B had a better inhibitory effect on Staphylococcus aureus than sample A and sample C.The inhibitory rates against Staphylococcus aureus were97.39% and 99.9% respectively at 5 mg/m L and 10 mg/m L drug concentrations.To explore the effect of its concentration on the activity of mouse glioma cells.The results showed that sample B had the best inhibitory effect on the growth of mouse glioma cells.When the drug concentration was 1 mg/m L,the addition of sample B could reduce the activity of mouse glioma cells to 14.50% of the original.In summary,this study systematically studied the extraction,separation and purification methods of flavonoids and saponins in muskmelon stems,leaves and seeds,formed an extraction process of flavonoids and saponins with high efficiency,high yield and high recovery rate,determined the appropriate purification process,and provided a more detailed technical scheme and basic data for the extraction and separation of flavonoids and saponins in muskmelon plants.The medicinal activity of purified muskmelon seeds in vitro was investigated to provide research basis and scientific basis for the rational development and utilization of natural active ingredient resources of muskmelon.更多还原