CN107252440A - 一种具有降血糖作用的山核桃叶总黄酮的制备方法和用途 - Google Patents
一种具有降血糖作用的山核桃叶总黄酮的制备方法和用途 Download PDFInfo
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Abstract
本发明涉及一种具有降血糖作用的山核桃叶黄酮提取物的制备方法和用途,本发明的山核桃叶总黄酮在各剂量均表现出良好的降血糖作用,对糖尿病具有显著作用,可用于制备预防或治疗糖尿病的药品、保健食品等。
Description
技术领域
本发明涉及中药技术领域,特别涉及一种具有降血糖作用的山核桃叶总黄酮的制备方法和用途。
背景技术
糖尿病是一组以高血糖为特征的代谢性疾病,临床上主要表现为慢性高血糖并伴有碳水化合物、脂肪、蛋白质代谢紊乱等。其病程较长,如果得不到及时有效的治疗,很可能引起全身多器官损害,出现感染、失明、肾病等并发症,严重危害人体健康。由于其广泛的发病人群及近年来逐渐上升的发病率,糖尿病已经成为继心血管疾病及癌症之后危害人类健康的第三大疾病。针对这种现状,大量降血糖药物随之问世,但目前临床上常见糖尿病治疗药物多为化学合成类药物,虽然疗效确切,也同时存在较多不良反应。天然药物,特别是我国传统中草药,在这种背景下具有得天独厚的优势条件,因此研究和开发高效低毒的天然药物,已成为预防和治疗糖尿病及并发症的热点。
山核桃树是胡桃科山核桃的叶子,具有清热解毒、杀虫止痒之效,在民间经常被于治疗脚趾湿痒、皮肤癣证等。此外文献研究表明山核桃叶提取物中含有大量的黄酮等抗氧化活性物质,具有抗肿瘤、抗衰老、抗诱变、抗血管生成等作用。而山核桃树本身是一类多年生落叶乔木,其叶在种植过程中常被丢弃,因此开发山核桃叶具有很大的经济价值和市场潜力。
发明内容
本发明的目的在于提供一种以黄酮类化合物为主要成分的山核桃叶总黄酮提取物;
本发明提供上述山核桃叶总黄酮提取物的制备方法;
本发明提供上述山核桃叶总黄酮提取物在预防或者治疗血糖升高症的药物或者保健品制备中的用途。
为实现上述目的,本发明采取的技术方案如下:
一种具有降血糖作用的山核桃叶总黄酮的制备方法,所述制备方法包括以下步骤:
(1)取山核桃叶40℃烘箱干燥,过40目筛,用95%乙醇提取在80-100℃常压提取30-90分钟,重复1-3次,过滤,合并滤液;
(2)滤液减压浓缩,离心取上清液,将此上清液过SD200大孔吸附树脂柱,上样速度为1BV/h-8BV/h,上样体积为4-8BV;在20-35℃,常压下,以1BV/h-8BV/h流速,用蒸馏水洗脱树脂,洗脱体积为5-10BV;在20-35℃,常压下,以1BV/h-8BV/h流速,用体积百分浓度为10-30%的乙醇水溶液洗脱树脂,洗脱体积为8-12BV;在20-35℃,常压下,以1BV/h-8BV/h流速,用体积百分浓度为40-90%的乙醇水溶液洗脱树脂,洗脱体积为8-12BV;收集40-90%乙醇洗脱液,浓缩,回收乙醇,减压干燥,得到山核桃叶提取物。
作为优选,步骤(1)中70-95%乙醇水溶液用量为每1g山核桃叶粉碎物加10mL。
作为优选,步骤(2)中离心参数为:2000-4000转每分钟,离心20分钟。
作为优选,步骤(2)中浓缩使滤液中乙醇浓度为45-65%。
一种具有降血糖作用的山核桃叶总黄酮的用途,将上述制备的山核桃叶总黄酮提取物可以与药剂或者食品上认可的辅料,采用现代制剂技术,制成以山核桃叶总黄酮为主的胶囊剂、片剂、散剂、口服液、软胶囊、丸剂、凝胶剂、缓释制剂等各种剂型;所述制剂用于临床治疗血糖升高以及制成含山核桃叶总黄酮的降血糖保健品。
本发明制备的山核桃叶总黄酮在降低血糖方面疗效显著,以四氧嘧啶致糖尿病小鼠模型实验结果显示,山核桃叶总黄酮具有显著的降血糖作用。
本发明的优点在于本发明采用方法简单经济,生产成本低,所得总黄酮含量较高且质量稳定。
具体实施方式
下面通过具体实施例,对本发明的技术方案作进一步的具体说明,但本发明不局限于这些实施例。
本发明中,若非特指,所采用的原料和设备等均可从市场购得或是本领域常用的。下述实施例中的方法,如无特别说明,均为本领域的常规方法。
实施例1
山核桃叶干燥后,机械粉碎,称取山核桃叶粉末200g,加10倍量(g:mL)的体积百分比为95%的乙醇溶液,提取2次,每次30min,合并滤液,在300±100pa真空度下减压浓缩,使滤液中乙醇浓度为65%,3000转每分钟,离心20分钟,取上清。称取SD200芳香族大孔吸附树脂(浙江争光实业股份有限公司)4.0g,放入层析柱(15mm×350mm i.d)中,以2BV/h的流速进行上样,上样体积为6.5BV。先用10BV水洗脱,再用10BV体积百分比为20%的乙醇溶液洗脱,最后用10BV体积百分比为70%的乙醇溶液洗脱,洗脱速度为4BV/h,收集70%乙醇洗脱液,减压旋转蒸发至干燥,获得山核桃叶提取物总黄酮,其中球松素、球松素查尔酮、白杨素及小豆寇明的总含量在55.3%以上。
实施例2
山核桃叶干燥后,机械粉碎,称取山核桃叶粉末400g,加10倍量体积百分比为95%的乙醇溶液,提取2次,每次30min,合并滤液,在300±100pa真空度下减压浓缩,使滤液中乙醇浓度为55%,4000转每分钟,离心20分钟,取上清。称取SD200大孔吸附树脂200.0g,放入层析柱(15mm×350mm i.d)中,依次用10BV水、20%乙醇、40%乙醇、70%乙醇洗脱,并且收集70%乙醇洗脱液,减压旋转蒸发至干燥,即得山核桃叶总黄酮提取物,其中球松素、球松素查尔酮、白杨素、小豆寇明的总含量在52.2%以上。
实施例3
山核桃叶干燥后,机械粉碎,称取山核桃叶粉末200g,加10倍量(g:mL)的体积百分比为95%的乙醇溶液,提取3次,每次90min,合并滤液,在300±100pa真空度下减压浓缩,使滤液中乙醇浓度为65%,2000转每分钟,离心20分钟,取上清。称取SD200芳香族大孔吸附树脂(浙江争光实业股份有限公司)4.0g,放入层析柱(15mm×350mm i.d)中,以8BV/h的流速进行上样,上样体积为8BV。先用5BV水洗脱,再以1BV/h流速用12BV体积百分比为30%的乙醇溶液洗脱,最后以5BV/h流速用10BV体积百分比为90%的乙醇溶液洗脱,收集90%乙醇洗脱液,减压旋转蒸发至干燥,获得山核桃叶提取物总黄酮,其中球松素、球松素查尔酮、白杨素及小豆寇明的总含量在54.3%以上。
球松素、球松素查尔酮、白杨素、小豆寇明的总含量测定方法参见沈勇等.高效液相色谱法测定山核桃叶中5个黄酮苷元含量[J].药物分析杂志,2013年05期。
将上述实施例1所得山核桃叶总黄酮对四氧嘧啶致糖尿病小鼠模型进行降血糖实验研究。
实验方法如下:
1实验材料与仪器
1.1实验动物
18~22g健康清洁级雄性ICR小鼠
1.2试剂与仪器
1.2.1试剂
山核桃叶总黄酮,CMC-Na、四氧嘧啶、二甲双胍,葡萄糖测定试剂盒、血清果糖胺测定试剂盒等。
1.2.1仪器
半自动生化仪;SENCOR-201旋转蒸发器;电子天平;离心机等。
2实验方法
将雄性ICR小鼠50只,随机分为6组,其中空白对照组10只,其余各组各8只,将40只模型小鼠禁食12h后尾静脉注射四氧嘧啶溶液80mg/kg造摸(正常对照组注射等量生理盐水),5d后测血糖,当血糖>11.1mmol/L时造模成功。然后将模型小鼠随机分为5组,分别为高血糖模型组,山核桃叶总黄酮50mg/kg低剂量组、100mg/kg中剂量组、200mg/kg高剂量组,50mg/kg二甲双胍阳性药组。实验期间正常对照组及模型组给予蒸馏水灌胃,其余各组按给药剂量灌胃,1次/d,连续14d。14d后,测定各组小鼠血糖、体重及血清果糖胺,并记录。实验结果均利用SPSS 18.0统计软件进行统计学处理。实验数据均以平均数±标准差表示,采用单因素方差分析,P<0.05时认为有显著性差异。
实验结果
表1山核桃叶总黄酮对高血糖模型小鼠的影响
注:与正常组比较,**P<0.01;与模型组比较,#P<0.05,##P<0.01。
表2山核桃叶总黄酮不同给药剂量对高血糖小鼠体重的影响
注:与正常组比较,**P<0.01;与模型组比较,#P<0.05。
表3山核桃叶总黄酮对高血糖模型小鼠血清果糖胺含量的影响
注:与正常组比较**P<0.01;与模型组比较#P<0.05;##P<0.01。
实验结果分析:本研究表明阳性药组、山核桃叶总黄酮低中高剂量组高血糖模型组小鼠相比,血糖水平均有显著下降,且血清果糖胺水平也显著降低并存在剂量依赖性,即实验结果表明山核桃叶总黄酮对四氧嘧啶致糖尿病小鼠模型具有显著降血糖作用。
以上所述的实施例只是本发明的一种较佳的方案,并非对本发明作任何形式上的限制,在不超出权利要求所记载的技术方案的前提下还有其它的变体及改型。
Claims (5)
1.一种具有降血糖作用的山核桃叶总黄酮的制备方法,其特征在于,所述制备方法包括以下步骤:
(1)取山核桃叶40℃烘箱干燥,过40目筛,用95%乙醇提取在80-100℃常压提取30-90分钟,重复1-3次,过滤,合并滤液;
(2)滤液减压浓缩,离心取上清液,将此上清液过SD200大孔吸附树脂柱,上样速度为1BV/h-8BV/h,上样体积为4-8BV;在20-35℃,常压下,以1BV/h-8BV/h流速,用蒸馏水洗脱树脂,洗脱体积为5-10BV;在20-35℃,常压下,以1BV/h-8BV/h流速,用体积百分浓度为10-30%的乙醇水溶液洗脱树脂,洗脱体积为8-12BV;在20-35℃,常压下,以1BV/h-8BV/h流速,用体积百分浓度为40-90%的乙醇水溶液洗脱树脂,洗脱体积为8-12BV;收集40-90%乙醇洗脱液,浓缩,回收乙醇,减压干燥,得到山核桃叶提取物。
2.根据权利要求1所述的一种具有降血糖作用的山核桃叶总黄酮的制备方法,其特征在于,步骤(1)中95%乙醇水溶液用量为每1g山核桃叶粉碎物加10mL。
3.根据权利要求1所述的一种具有降血糖作用的山核桃叶总黄酮的制备方法,其特征在于,步骤(2)中离心参数为:2000-4000转每分钟,离心20分钟。
4.根据权利要求1所述的一种具有降血糖作用的山核桃叶总黄酮的制备方法,其特征在于,步骤(2)中浓缩使滤液中乙醇浓度为45-65%。
5.一种具有降血糖作用的山核桃叶总黄酮的用途,其特征在于,权利要求1制备的山核桃叶总黄酮提取物可以与药剂或者食品上认可的辅料,采用现代制剂技术,制成以山核桃叶总黄酮为主的胶囊剂、片剂、散剂、口服液、软胶囊、丸剂、凝胶剂、缓释制剂等各种剂型;所述制剂用于临床治疗血糖升高以及制成含山核桃叶总黄酮的降血糖保健品。
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