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荷叶总黄酮提取纯化工艺及减肥降脂作用的研究
Separation and Purification of Total Flavonoids from Folium Nelumbinis and Its Weight-reducing and Lipid-lowering Effects
【作者】 薛冬娜;
【作者基本信息】 西南大学 , 植物学, 2008, 硕士
【摘要】 肥胖是一种以身体脂肪含量过多为重要特征的,多病因的,能够并发多种疾病的慢性病,与艾滋病、吸毒、酗酒并列为世界性四大医学社会问题,并可导致脂质代谢异常,胰岛素抵抗和高血脂、高血压、睡眠呼吸暂停症、脂肪肝、胆结石、脑卒,骨关节病、癌症等疾病,随着肥胖人口的日益增多,肥胖已经成为我国及广大发达及发展中国家面临的一个严重的公共健康问题。目前,治疗肥胖的西药品种繁多,疗效也较明显,但因其副作用强,不良反应多,大多不适于长期使用或已禁止使用。近几年来,中医学对肥胖病进行了广泛而深入的研究,也取得了较大的进展。又因为中医治疗肥胖症的副作用小、反弹率低,越来越受到国内外学者的重视。从中药提取物的国际市场现状来看,随着中药提取物在“替代疗法”方面的广泛应用,人们越来越意识到,许多传统的草药比临床使用的西药疗效更好,更具有安全性,国际市场需求不断增大。而我国植物提取物的年出口额在中药出口商品中的构成比呈明显的上升趋势,其国际市场空间也在不断扩大。荷叶系睡莲科植物莲的干燥叶,具有清热解暑,升发清阳,凉血止血之功效,其分布范围广泛,产量巨大。其主产地于中南地区及江浙一带,尤其是湖北省具有悠久的种植历史和丰富的栽培资源。荷叶含有槲皮素、荷叶碱等多种化学成分,被广泛应用于具有减肥降脂功能及治疗单纯性肥胖症的中药单复方制剂中,具有良好的临床效果。由于荷叶潜在的临床使用价值,加之鄂产荷叶来源广泛,质量良莠不齐,尚缺乏全面、统一的品质评价手段,给临床应用造成诸多不便。本文采用HPLC法测定槲皮素和荷叶碱的含量,同时采用亚硝酸钠-硝酸铝-氢氧化钠比色法和酸性染料比色法测定其总黄酮和总生物碱的含量,结果显示汉川、潜江、武汉汉阳荷叶的指标成分含量较高,九月为荷叶采收的最佳时期。通过多指标综合评价荷叶药材质量,弥补了《中国药典》中关于使用荷叶碱单一质控指标的局限性。在明确鄂产荷叶各有效成分含量的基础上,结合大孔吸附树脂技术提取分离其黄酮部位,以槲皮素为指标,采用正交试验优化黄酮提取物的工艺条件,选择适宜的树脂类型,并考察供试品溶液pH值、吸附流速、吸附液浓度、径高比、泄露曲线、清洗液pH值、洗脱液浓度、洗脱液用量等因素对黄酮部位分离的影响,结果表明:选择HPD100型树脂对荷叶黄酮组分进行吸附与解吸性能较好,采用吸附液浓度为0.2g·mL-1、流速为4BV·h-1的、吸附液pH为1~3、径高比为1:9、吸附液体积为柱床体积的11倍参数组合应用效果良好,清洗液pH小于5、2BV 50%乙醇洗脱效果较好且经济省时。按上述工艺流程分离制备荷叶黄酮提取物,通过建立大鼠营养性肥胖模型,考察荷叶黄酮提取物对于营养性肥胖大鼠及血脂指标的影响,结果表明,荷叶黄酮部位可明显抑制大鼠体重的增加,降低其Lee’s指数及脂肪重量,其中高剂量组能显著降低TC、TG水平,升高HDL-C/LDL-C,与模型组比较具有显著性差异。因此荷叶黄酮具有较好的减肥效果,而对动物体质耐力无明显影响。总之,通过鄂产荷叶中黄酮、生物碱化合物成分的分析以及大孔吸附树脂分离纯化黄酮工艺流程的优化,为评价荷叶减肥降脂相关活性提供了必要前提,从而为进一步阐明其分子机制提供了科学依据,并为今后建立化学指标和生物学指标相结合的质量评价体系奠定了基础,在确定荷叶品系品质的基础上对其进行规模化种植,开发新型具减肥降脂功效的保健食品,有利于加快鄂产荷叶的经济开发利用;同时增加中部地区莲农的经济收入,促进当地经济社会和谐发展。
【Abstract】 Overweight and obesity are defined as abnormal or excessive fat accumulation that presents a risk to health and are major risk factors for a number of chronic diseases.It is one of four biggest social and medical problems following AIDS,drug taking and excessive drinking.It can interferes with lipid metabolism,induces insulin resistance,hyperlipemia,hypertension,sleep apnea,fatty liver,gallstone,osteoarthrosis,cancer,etc.Nowadays,obesity patients are more and more,it has already been a significant public health problem that our country and many developed and developing countries are facing.At present,western medicines are numerous and effective,but they are not suitable for long-term use or has been forbided because of obvious side effect and untoward reaction.In recent years,therapy of obesity was studied extensively and profoundly in traditional Chinese medicine and has made important progress.Because of less side effects and low recurring rate,scholars all over the world have paied more and more attentions on obesity treatment with traditional Chinese medicine.Besids,because of vicarious treatment has been used widely in international market,more and more people notice that lots of traditional Chinese medicines are safer and more effective than western medicine.Its international market demand are widely increasing.In recent years,ratio of extract in export products of traditional Chinese medicine is ascending in our country.Its international market potential are extensive.Folium Nelurnbinis is the dried leaf of Nelumbo nucifera Gaertn.It has the effect of clearing away summer-heat and heat evil,ascending the lucid yang and cooling blood to stop bleeding.It grows in middle and south of China and the region of Zhejiang and jiangsu.Especially in Hubei province,it has a long history of cultivation and has abundance resources.Folium Nelumbinis contains kinds of chemical ingredients and has been added to single and compound recipes treating obesity clinically.Because of the potential clinical value of Foliurn Nelumbinis and the extensive resource,its quality is difficult to control.Being lack of unified estimated standard causes much inconvenience in clinical application.The thesis aims to establish methods with HPLC,Sodium Nitrite-aluminum nitrate-sodium hydroxide colorimetry and acid-dye colorimetry to determine the content of quercetin, nuciferine,total flavonoids and total alkaloids in Folium Nelumbinis respectively.The results show that the contents of index ingredients in areas of Hanchuan,Qianjiang and Hanyang were relatively higher than that from other habitats,and the month of September is the best time to collect.On the basis of confirming the index ingredients,the total flavonoids were extracted with macroporous adsorptive resins from Folium Nelumbinis.The process was optimized by employing orthogonal test design with the content of quercetin as the index.The resin HPD 100 had much better adsorption capacity and desorption ratio by using 2 BV 50%aqueous ethanol to extract the flavonoids from Folium Nelumbinis than other resins.The results showed that HPD100 possessed the highest absorption capacity under the concentration of sample 0.2g·mL-1,the condition of flowing rate of 4BV·h-1.Macroporous adsorptive resin HPD100 may be value of isolating and purifying the flavonoids from FoIium Nelumbinis.The flavonoids extract was prepared according to the technological process above,and the effect on the trophism obesity rat and blood-fat index was investigated.Tile result showed that the rats’ weight,Lee’ indexes,and the weight of rats’ fat was obviously descended.It also can reduced TC and TG,yet improve the HDL-C/LDL-C in blood serum.Thus it can be seen that the flavonoids extract of Folium Nelumbinis have significant effect on the reduction of fat and blood lipids.Generally speaking,through the analysis of flavones and alkaloids in Hubei province and optimized method of extraction of flavonoids with macroporous adsorptive resins,it supplied the essential precondition to evaluate the activity of anti-obesity and degrading the blood-fat with Folium Nelumbinis.Simultaneously,it is possible to elucidate its molecule mechanism with concerned experimental data.It established basis to set up the system of quality evaluation combining the chemistry and biology index.It is in favor of the promote of Folium Nelumbinis economy in Hubei province.It could develop the novel nutritious food and drinks,which will increase the income of peasants who plant Folium Nelumbinis.
【Key words】 Folium Nelumbinis; The flavonoids; Quercetin; Macroporous adsorptive resins; Obesity;
- 【网络出版投稿人】 西南大学 【网络出版年期】2011年 S2期
- 【分类号】R284;R285
- 【被引频次】5
- 【下载频次】517