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微波辅助提取、纯化葡萄籽中原花色素及其抗氧化性检验
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摘要
原花色素是一类具有抗氧化和清除自由基、保护心脑血管、防癌抗癌、保护肝脏、防治糖尿病和抗疲劳等作用的黄酮类化合物。在药品、化妆品、保健食品等领域有非常大的应用价值。葡萄籽是原花色素的重要来源。因此,如何更有效地从其中提取原花色素的研究开发具有非常重要的意义。
     以微波作为外加物理场,采用乙醇-水为提取剂时,利用单因素优选和正交实验筛选探索提取葡萄籽中原花色素的适宜提取工艺条件。通过实验发现适宜的提取条件为:提取剂乙醇的浓度为60%,料液比为1:8,微波功率为650w的条件下加热80s,然后在40℃下浸泡3小时,此时原花色素的提取率为4.52mg/g。利用5种大孔吸附树脂(S-8、NKA-9、AB-8、X-5、D4006)对原花色素进行静态吸附及解吸试验,筛选出AB-8型树脂是较适宜的填料。那么以AB-8型大孔吸附树脂为分离介质,完成原花色素的动态吸附和解吸试验,结果显示:原花色素水溶液浓度为2.5mg/ml,以2.0BV/h的流速流下,使树脂吸附达到饱和;解吸时先用水淋洗,解吸剂乙醇浓度为80%,以1.0BV/h的流速缓慢流下,解吸剂用量为1.0BV。
     确定邻苯三酚自氧化法测原花色素抗氧化活性的最佳操作条件为:在温度25℃时,配置pH=8.2的Tris-HCl溶液作为缓冲体系,邻苯三酚浓度为30mmol/L,测定波长为420nm。
     葡萄籽中的原花色素对油脂的自由基氧化反应有抑制作用。通过大孔树脂吸附的精制物的抗氧化性能明显的优于未精制物。可见,通过大孔树脂的吸附分离作用除去杂质后,有利于原花色素发挥其抗氧化作用。
Proanthocyanidin is a kind of flavonoid chemical which has function ofantioxidation, clearance of free radicals, protection of blood vessel, cancer preventionand anticancer, anti-inflammatory, liver protection, diabetes prevention andanti-fatigue. It plays a key role in the field of pharmaceutical, cosmetic and healthproducts. Grape seed is an important resource of proanthocyanidin, so that study onthe extration of proanthocyanidin has significant meanings.
     This paper uses microwave as external field. Ethanol-water solution is used asextracting agent. Single element optimized method and orthogonal test method areadopted to select and confirm the optimal extracting condition. According toexperiment, the optimal extracting condition is that concentration of 60% ethanolsolution and 1:8 of feed liquid ratio and heating by 650w microwave for 80 secondsand soaking for 3 hours in 40℃circumstance.The yield of proanthocyanidin canreach to 0.452mg/ml
     State absorption and desorption test of proanthocyanidin are done on five kindsof macropore adsorption resins (S-8、NKA-9、AB-8、X-5、D4006) to study theadsorbability of resins above. AB-8 resin is selected as optimal packing. The test onAB-8 indicates that the resin saturates on the condition of 2.0BV/h and 2.5 mg/ml androom temperature. During desorption process, resin is washed by water firstly andthen 80% ethanol solution as desorption agent under flow rate 1.0BV/h. Totalconsumption of desorption agent is 1.0BV.
     The optimum operation condition using 1,2,3-trihydroxybenzene autooxidationmethod to test inoxidizable activity is achieved that using 420nm length measurewave under 25℃, pH 8.2 Tris-HCl solution is used as buffer system; concentration of1,2,3-trihydroxybenzene solution is 30mmol/L.
     Depression function of proanthocyanidin on vegetable oil autooxidation is alsostudied in this paper. The result shows proanthocyanidin which extracted from grapeseed has the depression function on free radical oxidation reaction of oil and grease.Inoxidizability of refined product by macropore resin absorption is much better thancrude product. Therefore, separation and purification of impurity through macroporeresin improves the inoxidizable capability of proanthocyanidin
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