【作者】 杨阳；

【导师】 李文雍； 孙敏；

【作者基本信息】 安徽大学 ， 细胞生物学， 2010， 硕士

【摘要】 目的：优化香椿总黄酮的提取工艺,比较太和香椿五个品种的总黄酮含量。建立太和香椿中槲皮素含量的测定方法,比较太和香椿四个品种的槲皮素含量。测定太和香椿腌制前后总黄酮含量及营养成分的变化。观察槲皮素对人胚肺二倍体成纤维细胞(MRC-5)中超氧化物歧化酶(SOD)活性、丙二醛(MAD)含量的影响。方法：1、设计正交实验对提取总黄酮的酶解水提法进行优化,以浸提液中总黄酮得率作为考察指标。采用最佳提取方法比较太和香椿五个品种的总黄酮含量。2、采用高效液相色谱法检测槲皮素含量,色谱柱为shim-pack:vp-ODS柱,流动相为甲醇-水(1:1,稀磷酸调pH为2.50),检测波长为368nm,流速为1.0ml·mim-1。3、以新鲜太和香椿和腌制7天的太和香椿作对照。分别采用乙醇浸提法、超声波辅助法、酶解水提法三种不同提取方法,以浸提液中总黄酮得率作为考察指标,比较香椿腌制前后总黄酮含量。分别采用考马斯亮蓝法、硫酸苯酚法、氧化还原滴定法,以蛋白质含量、可溶性糖含量以及维生素C含量为检测指标,比较太和香椿腌制前后营养成分含量。4、用槲皮素对MRC-5进行处理,用羟胺法检测细胞中SOD含量,比色法检测细胞中MAD含量。结果：1、酶解水提法的最佳提取条件是酶用量15 U/g,酶解时间90 min,酶解后水提时间120 min,料液比1：40。五个品种中红油椿的总黄酮含量最高,黑油椿次之,其次是红椿和青椿,毛椿的总黄酮含量最少。2、太和香椿中槲皮素浓度在83.2～187.2μg·ml-1范围内线性关系良好,r=0.9995。槲皮素含量最高的为红椿,其次是红油椿,黑油椿槲皮素含量略低于红油椿,含量最低的为青椿。3、三种提取方法均表明腌制七天后香椿总黄酮含量略有降低,维生素C与蛋白质含量下降明显,而可溶性糖含量明显上升。4、与正常对照组相比,用药组MRC-5中SOD活性明显增加,MDA含量明显下降。结论：1、酶解水提法是一种提取太和香椿中总黄酮的较优方法。太和香椿的总黄酮含量比较高,具有非常高的药用和食用价值。2、本实验采用的高效液相色谱法回收率、精密度良好,操作简单,适用于太和香椿中槲皮素含量的测定。3、香椿腌制七天后,其总黄酮与其它营养成分均受到不同程度的破坏,其营养价值远低于新鲜的香椿,该实验数据可为以后研究香椿的食用及保存方法提供有价值的参考数据。4、槲皮素具有较强的抗氧化作用。更多还原

【Abstract】 Purpose:To optimize the extraction process of total flavonoids of Toona sinensis and to compare the content of total flavonoids in Toona sinensis among five defferent species cultivated in Taihe county.To establish HPLC method for the determination of quercetin in Toona sinensis cultivated in Taihe county. To compare meletin of Toona sinensis among four defferent species cultivated in Taihe county.To determine the content of total flavonoids and nutrient component in fresh and pickled Toona sinensis. To observe effects of quercetin on activity of SOD and MDA of human fetal lung diploid fibroblasts (MRC-5).Methods:1. Designing the orthogonal experiment optimizes Enzyme treatment,by referring the yield of total flavonoids in extraction.Use the best extraction method to extract the total flavonoids from five species of Toona sinensis cultivated in Taihe county,and compare the size of its content.2. The HPLC analysis was performed on a shim-pack:vp-ODS column with methanol-water(1:1,adjust pH to 2.50 by dilute phosphoric acid) as the mobile phase,the detective wavelength was 368nm,the flow rate was 1.0ml·mim-1.3. Using three different methods:ethanol ertraction,ultrasonic ertraction,enzyme treatment to determine the content of total flavonoids in fresh and seven-day pickled Toona sinensis with the yield of total tlavonoid as examine items.using coomassie brilliant blue method, phenol sulfuric acid method and redox titration,with the content of protein, soluble sugar and Vc as examine items,to determine the content change of nutrient component in fresh and seven-day pickled Toona sinensis.4. MRC-5 cell lines were treated with quercetin.Using hydroxylamine ethod and chemical colorimetry method to determine the contents of SOD and MDA.Results:1. Here are the best extraction condition of Enzyme treatment. The dosage of enzyme is 15 U/g, the time of Enzymolysis is 90 min, the time of the extraction after enzymolysis is 120 min, and the ratio between solid and liquid is 1:40. The total flavonoid content of red oil toona is the highest, followed by black oil toona, red toona, green toona, and hairy toona is the least.2. Quercetin at the concentrations of 83.2～187.2μg·ml-1 showed good linearity(r=0.9995). The quercetin content of red toona is the highest, followed by red oil toona, black oil toona, and green toona is the least.3. The analysis show a significantly decrease of protein and and Vc in seven-day pickled Toona sinensis,the decrease of total flavonoids isn’t so significantly.;And the increase of soluble sugar is also not so significantly.4. Compared with those in the control group, the activity of SOD increased obviously, and the content of MDA were decreased significantly in the drug group.Conclusion:1. Enzyme treatment is a better method of total flavonoids extraction. The total flavonoid content of Toona sinensis cultivated in Taihe county is relatively high, with very high value.2. The HLP method was simple,sensitive and reliable.lt can be uesd for the content determination of quercetin.3. After be pickled,the total flavonoid and nutrient component in Toona sinensis were varyingdegrees damged,it’s nutritional value were significantly lower than fresh Toona sinensis.This experimental data can be a valuable reference for the study of eating and preservation methods of Toona sinensis.4. Quercetin has a strong antioxidant properties.更多还原