【作者】 殷志萍；

【导师】 赵文；

【作者基本信息】 河北农业大学 ， 农产品加工及贮藏工程， 2012， 硕士

【摘要】 本文以张家口产的蔷薇红景天根茎为原料,研究蔷薇红景天原花青素分离纯化工艺,并对其抗氧化和抗肿瘤活性进行了初步研究,为进一步开发该新食物资源提供依据。研究方法:(1)蔷薇红景天原花青素纯化工艺研究。首先,采用醇提法获得蔷薇红景天乙醇提取物,再通过乙酸乙酯萃取的方法分离蔷薇红景天原花青素低聚物与高聚物,最后,采用AB-8大孔吸附树脂吸附的方法,对上述两种原花青素粗品进一步分离纯化。在前期考察了上样液浓度、上样流速、解析液浓度等条件的基础上,本研究对AB-8大孔吸附树脂的上样液pH、解析液pH和径高比进行筛选,优化AB-8大孔吸附树脂的纯化工艺,筛选最佳吸附、解析条件,制备较高纯度的蔷薇红景天原花青素低聚物和高聚物。(2)蔷薇红景天原花青素的抗氧化性研究。通过体外实验研究蔷薇红景天原花青素对羟自由基、DPPH自由基、邻苯三酚自氧化产生的超氧阴离子、亚硝酸根离子的清除作用;利用整体动物实验观察蔷薇红景天原花青素低聚物对小鼠外周血、肝脏、心脏和脑组织中抗氧化酶(SOD,GSH-Px)和脂质过氧化物(MDA)的影响。(3)蔷薇红景天原花青素低聚物抗肿瘤活性研究。建立肉瘤-180(S180)荷瘤动物模型,观察S180在动物体内的生长情况,根据平均瘤重计算抑瘤率。研究结果:(1)乙酸乙酯萃取最佳工艺为萃取时间25 min、体积比(乙酸乙酯︰提取液)1.5︰1、萃取次数4次;AB-8大孔吸附树脂的最佳上样液pH为4.5,解析液pH为5,径高比为1︰40(cm)。由此分离纯化得到蔷薇红景天原花青素低聚物的纯度为88. 3%,而蔷薇红景天原花青素高聚物的纯度为98%。(2)蔷薇红景天原花青素具有较强的清除DPPH自由基、羟自由基和亚硝酸根离子的能力,且清除效果均优于抗坏血酸。经蔷薇红景天原花青素低聚物处理的小鼠外周血、心脏、肝脏、脑组织中SOD、GSH-Px活性有所增加,MDA含量显著下降; 200、400 mg/kgbw剂量组可明显提高上述组织中GSH-Px活性,降低MDA含量,与对照组相比均有显著性差异(P<0.05),三个剂组的外周血、心脏、脑组织中SOD活性明显高于对照组(P<0.05),肝脏SOD活性与对照组比较差异不显著(P>0.05)。(3)蔷薇红景天原花青素低聚物具有一定的抗肿瘤活性,且雌雄小鼠之间存在差异。低、中、高三个剂量组雄性动物的平均瘤重与模型对照组比较均明显降低,且有显著性差异(P<0.05),其抑瘤率分别为47.5%、51.4%、56.9%;而雌性动物的平均瘤重较模型对照组有所降低,但统计学处理无显著性差异,抑瘤率分别为24.4%、40%、41.1%。结论:本研究采用乙酸乙酯萃取与AB-8大孔吸附树脂吸附相结合分离纯化蔷薇红景天原花青素,得到原花青素产品纯度较高,效果良好;蔷薇红景天原花青素具有良好的自由基清除作用,其低聚物能有效对抗动物机体的脂质过氧化,并且具有一定的抗肿瘤活性。更多还原

【Abstract】 In order to provide theoretical foundation for developing new food resource, the present paper used rhizome of Rhodiola rosea as raw material to study separation and purification techniques of proanthocyanidins, and also conducted assays investigating its anti-oxidant functions and anti-tumor effects.The main researches are as follows:(1) Study of purification of Rhodiola rosea proanthocyanidins. Firstly, Rhodiola rosea was extracted by ethanol, before the extract was purified by acetic ether leaching and AB-8 macro porous resin absorption. Based on former researches about sample concentration, flow velocity of sample loading and concentration of desorption solution, the present study optimized purification technique of AB-8 macro porous resin absorption as the sample liquid pH、desorption solution pH and diameter to height ratio. The optimized conditions of leaching, absorption and desorption were explored, and samples of oligomer and polymer with high purity were achieved.(2) Researches on anti-oxidative effects of Rhodiola rosea proanthocyanidins. The eliminating effects on·OH,·DPPH,·O2 produced by Pyrogallol auto-oxidation and NO2- of Rhodiola rosea proanthocyanidins were evaluated by tests in vitro. And integral animal experiments were employed to observe Rhodiola rosea Proanthocyanidin oligomers’influence on SOD, GSH-Px,and MDA levels of peripheral blood, liver, heart and brain.(3) Study on anti-tumor property of Rhodiola rosea proanthocyanidins. The S180 tumor–bearing animal model was established, and we monitored the development state of sarcoma in animal body; According to the average tumor heavy,the inhibitory rate (refers to Rhodiola rosea proanthocyanidins’function on inhibiting tumor growth) was calculated.The main results are along these lines:(1) The optimized conditions of acetic ether leaching were: extracting time of 25 min, volume rate (acetic ether to extractant) of 1.5:1, operation times of 4. The optimized conditions of AB-8 macro porous resin absorption were: sample liquid pH of 4.5, desorption solution pH of 5, and diameter to height ratio of 1:40 (cm). Under the above optimized conditions, Rhodiola rosea Proanthocyanidin was separated and refined, with product purity reaching 88. 3%, and polymer product purity was 98%.(2) Rhodiola rosea proanthocyanidin had strong ability of diminishing·OH, DPPH·,·O2 and NO2-, and the outcome was better than ascorbic acid. After administration of Rhodiola rosea proanthocyanidins, activities of SOD and GSH-Px in mouse blood, heart, liver and brain improved, while MDA content declined; 200 and 400 mg/kgbw dose groups notably increased GSH-Px activity and decreased MDA content in the mentioned 4 tissue samples, as significant differences were observed in comparison to the control group(P<0.05); however, SOD activity in liver showed no significant difference between dose groups and control group(P>0.05), while in the other 3 tissue samples, SOD activities in dose groups were significantly different from the control group(P<0.05).(3)Rhodiola rosea proanthocyanidin oligomer unfolded some degree of anti-tumor activity, which was dissimilar among mice of different genders. The average tumor weights of male mice from low, middle and high dose groups obviously reduced, and displayed significant differences in contrast to the control group, the inhibitory rates (refer to Rhodiola rosea proanthocyanidins’function on inhibiting tumor growth) were 47.5%, 51.4% and 56.9% separately. However, though average tumor weights of female mice from dose groups were less than the control group, no significant difference was revealed in the data, and inhibitory rates were 24.4%, 40% and 41.1% separately.The results indicated that the separation method of acetic ether leaching along with AB-8 macro porous resin absorption had admirable effects, and Rhodiola rosea proanthocyanidin products were with high purity. Through the anti-oxidative experiments and anti-tumor explorations, Rhodiola rosea proanthocyanidin oligomer was proved to be an excellent natural antioxidant with some degree of anti-tumor activity.更多还原