【作者】 高波；

【导师】 卢卫红；

【作者基本信息】 哈尔滨工业大学 ， 食品科学与工程， 2013， 硕士

【摘要】 孔石莼是一种大型海洋经济绿藻，在我国资源十分丰富。其味道鲜美，具有很高的营养价值与药用价值。近些年来，孔石莼的工厂化育苗与人工养殖技术日趋完善，但关于它生物活性的研究还处于起步阶段，远远跟不上养殖栽培的步伐，尤其是抗氧化生物活性，亟待开展研究工作对其进行系统评价。本论文选取孔石莼作为研究对象，以自由基清除能力为体外抗氧化评价指标，筛选、分离、鉴定了孔石莼中的抗氧化物质，以期将孔石莼抗氧化活性的研究成果应用到天然抗氧化剂、疾病治疗药物的开发上来。本研究主要取得如下研究成果：(1)采用超声辅助提取技术，用乙醇水溶液对孔石莼进行了提取，研究考察了浸提温度、浸提料液比、乙醇浓度、超声时间对可溶性固形物得率的影响，通过一系列单因素试验及正交试验分析，对孔石莼粗提物的提取条件进行了优化，并对优化结果进行了验证。结果表明，最佳的提取条件为：浸提温度60℃、浸提料液比1:25g/ml、乙醇浓度60%、超声时间2h；在此条件下，孔石莼中可溶性固形物的平均得率为11.81%，相对标准偏差(RSD)为0.53%，表明结果具有较高的精密度与可靠性。(2)采用液—液萃取方法对孔石莼的粗提物进行初步分离，以ABTS自由基清除能力、羟基自由基清除能力、DPPH自由基清除能力为检测指标进行综合分析。结果显示，正丁醇层具有较好的抗氧化能力。实验选用D101型大孔树脂对正丁醇层做进一步的分离纯化，用不同浓度的乙醇溶液进行梯度洗脱，洗脱组分检测其对自由基的清除能力，最终筛选出抗氧化能力较强的SR-50组分作下一步的研究分析。为了得到较纯的目标产物，研究首次应用中压液相色谱快速纯化工艺，系统分离纯化了SR-50组分，在最适的洗脱条件(氯甲比=94:6)下，纯化得到了SR-50-I组分，进行相应抗氧化能力的分析，发现SR-50-I组分对ABTS自由基、羟基自由基、DPPH自由基的清除作用均强于SR-50组分，说明经过中压液相色谱的纯化，物质中抗氧化活性成分的纯度提高了。(3)联合应用紫外光谱分析、常规显色反应及液质联用方法对纯化所得的抗氧化有效组分SR-50-I组分进行鉴定，结合文献资料，初步推断了SR-50-I组分含有儿茶酚、表儿茶素和桑色素三种多酚、黄酮类化合物。此三种化合物有望成为标识成分作为孔石莼中抗氧化活性物质的质量控制指标。更多还原

【Abstract】 As a kind of large ocean economic green alga, Ulva pertusa is abundant in China,containing both high nutritional and pharmaceutical value. Recently, the industrializedseedling production and manual cultivation technology of Ulva pertusa has been welldeveloped, but study about its biological activity is still in the initial stage, far less thanthe pace of Aquaculture cultivation, especially its antioxidation feature has yet to bepromoted.Based on Ulva pertusa, indicated by its antioxidation eliminating free radical, thisresearch selected, separated, and identified antioxidants in Ulva pertusa, wishing toapply achievement of its antioxidation into natural antioxidant, curing medicines. Themain results are as follows:With ultrasound technique, crude ingredients in Ulva pertusa was extracted, thenexamined influence of temperature, solid-to-liquid ratio, concentration of ethanol, timeto soluble solid material, through series of single factor test and orthogonal test,extraction condition was optimized. Results showed that the best extraction system is:temperature60℃, solid-to-liquid ratio1:25, concentration of ethanol60%, time ofultrasound2h. Under this circumstance, extraction rate of soluble solid matter in Ulvapertusa is11.81%, RSD is0.53%, indicating that results is highly accurate andreliable.By way of liquid-liquid extraction, indicated by elimination of ABTS free radicals,hydroxyl free radical, DPPH free radical, crude extraction of Ulva pertusa was studied.Results showed that extraction by normal butanol has better antioxidation. Then D101macroporous resin was adopted for further purification, eluded with ethanol in differentconcentration. Elution part was examined with antioxidation ability. Finally, SR-50possessing higher antioxidation ability was selected for further research. In order toobtain purer product, the study employed medium pressure liquid chromatography forthe first time to purify SR-50systematically. Under most suitable elutioncondition(chloroform:methanol=94:6), SR-50-I was obtained, then according toantioxidation analysis, the ability of elimination of ABTS free radical, hydroxyl freeradical, DPPH free radical were much stronger than that of SR-50. which means purityof antioxidation in sample purified was enhanced.Combined with Ultraviolet Spectrum Analysis, Conventional color reaction andLC-MS, purified constituent SR-50-I was identified, and preliminarily conclude thatSR-50-I contains catechol, epicatechin, morin.They are expected to be identified asquality control indedx of Ulva pertusa antioxidant ingredient.更多还原