【作者】 潘一峰；

【导师】 瞿伟菁；

【作者基本信息】 华东师范大学 ， 植物学， 2005， 硕士

【摘要】 在国家实施天然林保护工程后,越桔（Vaccinium vitis-idaea L.）已成为林业系统利用野生资源发展果用产业（饮料、酿酒等）循环经济的经济林种之一。与此同时,初级利用后的生产弃物越桔果渣的深度开发不仅有助于防止工业弃物的二次污染,也有助于延长资源加工的产业链。 为此,本文首先对越桔果渣的成分进行了分析,并采用高效液相色谱法对越桔果渣中黄酮甙元进行定性和定量分析。HPLC用甲醇和水（60:40）作为流动相以0.8mL/min流速进行洗脱,368nm波长检测越桔果渣水解液中黄酮类成分并测定其含量。结果发现,越桔果渣中黄酮甙元主要有杨梅素和槲皮素,其含量分别为0.020%和0.069%,提示越桔果渣具有深度利用的潜力。 在此基础上,为实现果用越桔残渣中次生代谢产物的深度开发,研究接着对越桔果渣总黄酮（FFV）的提取纯化工艺进行了试验。以乙醇的浓度、时间、温度、以及pH为考察指标,在单因素水平研究了FFV的提取方法,然后用正交法确定了较好的提取工艺;比较不同大孔树脂（D101、HPD600、AB-8、NAA-9、X-5和D4020）对FFV的吸附性能,通正交试验确定大孔树脂吸附纯化FFV的最佳条件。结果表明:FFV的最佳提取工艺为:乙醇的浓度60%,时间2h,温度60℃,液料比1:20,提取次数3次。HPD600大孔树脂对FFV的吸附性能最好,HPD600大孔树脂对FFV吸附纯化的最佳条件为60%乙醇作为洗脱剂,树脂投入量与黄酮含量比值10g:167.5mg,洗脱速度1mL/min,吸附时间1h。总得率约为3.89%,通过HPLC表征分析,FFV中杨梅素和槲皮素的黄酮甙元含量约为6.42%。 为证明越桔黄酮的生理活性,本文在提取纯化获得成功的基础上,继续就同类研究尚未涉及的领域,FFV对糖脂代谢的主要影响进行了深入的研究。 首先研究了越桔果渣黄酮类成份的抗氧化活性。结果表明FFV可有效降低大鼠肝肾中自发性和Fe²⁺-Vitc、H₂O₂诱导的MDA生成,抑制H₂O₂诱导产生的红细胞溶血和肝脂质过氧化作用,以及阻止Fe²⁺诱导卵黄蛋白低密度脂蛋白的氧化作用,显著降低小鼠的肝、肾、脑的MDA含量,提高肝和血浆中SOD和CAT酶的活性。 试验进一步从整体水平上研究了FFV对糖脂代谢的影响。在灌胃8d、16d、24d、32d时测正常小鼠的血糖、血脂水平;在20d时进行糖异生实验。结果表明FFV可显著地降低小鼠血清总胆固醇水平,抑制对小鼠糖异生作用,对血糖有一定程度的调节作用,而不影响甘油三酯的水平。 为此,研究采用Alloxan致高血糖模型小鼠研究了FFV对糖代谢异常的治疗更多还原

【Abstract】 Since the implementation of Natural Forest Protection Program, Vaccinium vitis-idaea L. has became one of the economic forest species of recycle economy developing fruit-using industries （drink, brew etc.） with wild resources in forestry system. Meanwhile, the deep development of the fruit residue of Vaccinium vitis-idaea L. （FRV） after primary using will help to prevent secondary pollutant of industry offal, and prolong the industry chain of resource processing.So, this paper firstly analyzed the components in FRV and did qualitative and quantitative analysis of the flavonoid aglycones in FRV by HPLC. Results showed , Two types of flavonoid aglycones, myricetin （0.020%） and quercetin （0.069%）, were detected in FRV by HPLC using methanol and water （60:40） as mobile phase at flow rate of 0.8 mL/min. The peak was detected at 368nm. This prompted that FRV had the potential for deep using.On this basis, for realization of deep development of secondary metabolic product in FRV, The optimum conditions of extraction and purification of flavonoids from FRV were studied in this article.Considering the concentration of ethanol, time, temperature and pH as indexes, we studied the extract means of flavonoids, then the optimum conditions of extract was selected by orthogonal design; Adsorption capacity of six macroporous resins （D101、 HPD600, AB-8 NAA-9、X-5 and D4020） for total flavonoids were compared. The optimum conditions of adsorbing and purifying total flavonoids from fruits residue with macroporous resin absorbent was selected by orthogonal design. The results showed that ethanol concentration 60%, time 2h, temperature 60℃,w/v 1:20, times 3 were the optimum extract conditions of FFV. The HPD600 resin was best one in the six macroporous resins. The optimum process condition was 60% ethanol as eluting solvent for l0g wet resin/167.5mg total flavonoids, flowing rate lmL/min, absorbing time for lh, the total yield of FFV was 3.89%. The flavonoidaglycones contents of myricetin and quercetin were 6.42% with the characterization analysis of HPLC .In order to identify the physiological activity of FFV on the basis of extraction and purification, this paper continued to study for the effect of FFV on glycometabolism and lipid-metabolism which were not being concerned in the same field of homologous studies.Anti-oxidation of FFV was researched, the results in vitro showed that FFV reduced MDA germination in the liver and kidney spontaneity or induced by the Fe²⁺-Vitc、 H₂O₂; restricted erythrocyte hemolysis induced by H₂O₂ or lipids peroxidation in the rat liver; and inhibited oxidation of low density lipoprotein （LDL） induced by Fe²⁺. The results in vivo showed that FFV reduced MDA germination in mouse liver kidney and brain spontaneity, improved the activity of SOD and CAT in liver and serum.The effect of FFV on glycometabolism and lipid-metabolism in normal mouse was investigated. The levels of serum glucose, serum cholesterol（TC） were determined when it lasted 8d,16d,24d and 32d, After 20d glyconeogenesis test was made. The Results showed, FFV reduced the levels of serum cholesterol significantly and serum glucose to some extent, restricted the gluconeogenesis and did not reduce the level of triglyceride（TG） in normal mice.Therefore, alloxan-induce diabetic mice were administered orally with FFV, the results showed that the glucose level of oral administration of 200mg/kg FFV to mouse exhibited significant reduction 18.05% on 7th d, and exhibited reduction 15.04% on 14th(1 and 15.27% on 21thd; 50mg/kg FFV had less reduction and no statistical difference; The effect of control drug was more strong than FFV. 200mg/kgFFV reduced the content of TG and TC, and reduced MDA germination in serum, improved the activity of SOD in liver and serum significantly, and 50mg/kg FFV had not the same effects.Preventive test of alloxan-induce diabetic mice showed that FFV could significantly inhibit the levels of fructosamine and serum glucose and improve the levels of SOD in serum and pancreas.The effect mechanism of FFV on abnormal gl更多还原