CN105596429B - 一种防治ii型糖尿病的莲子心总黄酮的制备及其应用 - Google Patents
一种防治ii型糖尿病的莲子心总黄酮的制备及其应用 Download PDFInfo
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Abstract
本发明公开了一种莲子心总黄酮的制备方法及其在辅助治疗II型糖尿病中的应用,属于医药保健品技术领域。其采用微波‑超声波协同辅助萃取法和超临界流体萃取法联合获得莲子心总黄酮,其提取率可达2.66%,将所得莲子心总黄酮混入辅料后,可制成药剂学上接受的剂型。经动物药效学验证,所得莲子心总黄酮可降低由链脲佐菌素(STZ)致病的II型糖尿病模型小鼠的血糖,降糖率可达到43.02%,并具有改善胰岛素抵抗的功效,能够降低II型糖尿病空腹血糖值,预防和辅助治疗II型糖尿病。
Description
技术领域
本发明属于医药保健品技术领域,特别涉及一种莲子心总黄酮的制备方法及其在辅助治疗II型糖尿病中的应用。
背景技术
糖尿病是由于胰岛素分泌缺陷及(或)其生物学作用障碍引起的以高血糖为特征的代谢性慢性疾病,其给人类健康造成的威胁仅次于心脑血管病和肿瘤。糖尿病已不仅仅是工业化发达国家的富贵病,亚洲包括中国在内的发展中国家已经成为糖尿病的重灾区,中国糖尿病人群数量占据全球患病人数的1/3,城市发病率为11.4%,总数为全球第一。糖尿病复杂的发病过程使人类至今尚未找到根治的方法,患者需要终身接受治疗。目前用于糖尿病临床治疗的药物以西药为主,虽然其具有作用效果明显、作用时间短的特点,但有一定副作用,易引起机体的不良反应,长期大剂量的服用更会损害人体机能。具有预防和辅助降血糖作用的保健品,在有效控制血糖和糖尿病的同时能降低副作用的影响,近年来成为人们消费的热点。
莲子心有广泛的药理作用,在传统中医药中占有重要地位,为《中国药典》所收录,且在历史中也早有记载,如唐代医家陈仕良所著的《食性本草》(约 937~957年),描述莲子心“生取为末,以米饮调下三钱,疗血渴疾、产后渴疾”;清代医家叶天士(1667~1746年)在《本草再新》中记录莲子心有“清心火,平肝火,泻脾火,降肺火。消暑除烦,生津止渴,治目红肿”之功效;此外,中医温病学家王士雄(约1808~1868年)于《随息居饮食谱》中对莲子心的药理作用亦有如下记载:“敛液止汗,靖热养神,止血固精”。黄酮为莲子心的主要活性成分,是莲子心广泛药理作用的主要物质基础。目前尚未见到利用微波-超声波协同辅助提取莲子心黄酮的工艺,及对莲子心总黄酮降血糖活性方面的研究。本发明对莲子心黄酮类化合物进行提取与富集,并经药效学验证,其能够显著降低II型糖尿病空腹血糖值,具有预防和辅助治疗II型糖尿病的功效,极具开发降糖保健食品的潜力。
发明内容
本发明的目的在于提供一种莲子心总黄酮的制备方法及其在辅助治疗II型糖尿病中的应用,所得莲子心总黄酮与临床治疗的药物配合使用,能有效预防和辅助治疗II型糖尿病,并降低药物的毒副作用。
为实现上述目的,本发明采用如下技术方案:
一种防治II型糖尿病的莲子心总黄酮的制备方法包括如下步骤:
1)称取新鲜莲子心,洗去杂质,40-50℃烘干至恒重后,超微粉碎至细度为10-15 μm;
2)向步骤1)制备的莲子心粉末中加入乙醇溶液,微波-超声波协同辅助提取5-15min,过滤;将滤渣按上述步骤重复提取2-3次,合并滤液,于3000-4000 r/min条件下离心10-20 min,取上清液,在50-65℃浓缩至浸膏状,冷冻干燥后得莲子心总黄酮粗品;
3)将步骤2)将制得的莲子心总黄酮粗品投入超临界萃取釜,将质量浓度为95%的乙醇溶液和液态CO2按体积百分数之比为1-10:90-99作为萃取剂通入萃取釜进行分离后,将所得萃取液真空浓缩即得莲子心总黄酮。
步骤2)中所用莲子心粉末重量与乙醇溶液的体积之比为1:20-40,所述乙醇溶液的质量浓度为70%-80%;
所述微波-超声波协同辅助提取中微波功率为300-400 W,超声功率为200-400 W,超声程序为运行1s后停止0~2s,如此循环。
步骤3)所述液体CO2的纯度大于99.99%。
所得莲子心总黄酮可与常规辅料配合,用于制备辅助治疗II型糖尿病的药物或保健品;所述药物或保健品为药剂学上接受的剂型,包括胶囊剂、片剂、颗粒剂、口服液。
本发明的显著优点在于:本发明将微波-超声波协同辅助萃取法和超临界流体萃取法联合运用,获得莲子心总黄酮,其提取率可达2.66%。本发明工艺稳定可靠,适合于工业化生产,所得莲子心总黄酮与临床治疗的药物配合使用,能有效预防和辅助治疗II型糖尿病,并降低药物的毒副作用。
具体实施方式
为了使本发明所述的内容更加便于理解,下面结合具体实施方式对本发明所述的技术方案做进一步的说明,但是本发明不仅限于此。
实施例1
一种防治II型糖尿病的莲子心总黄酮的制备方法包括如下步骤:
1)称取新鲜莲子心,洗去杂质,40℃烘干至恒重后,超微粉碎至细度为10μm;
2)在步骤1)制备的莲子心粉末中按质量体积比1: 20(g/mL)加入质量浓度为70%的乙醇溶液,微波-超声波协同辅助提取15 min,微波功率为300 W,超声功率为200 W,超声程序为运行1s后停止0~2s,如此循环,然后过滤;将滤渣按上述步骤重复提取2次,合并滤液,于3000 r/min条件下离心20 min,取上清液,在50℃浓缩至浸膏状,冷冻干燥后得莲子心总黄酮粗品;
3)将步骤2)将制得的莲子心总黄酮粗品投入超临界萃取釜,将质量浓度为95%的乙醇溶液和纯度大于99.99%的液态CO2按体积百分数之比为1:99作为萃取剂通入萃取釜进行分离后,将所得萃取液真空浓缩即得莲子心总黄酮。
实施例2
一种防治II型糖尿病的莲子心总黄酮的制备方法包括如下步骤:
1)称取新鲜莲子心,洗去杂质,45℃烘干至恒重后,超微粉碎至细度为10μm;
2)在步骤1)制备的莲子心粉末中按质量体积比1: 30(g/mL)加入质量浓度为75%的乙醇溶液,微波-超声波协同辅助提取10 min,微波功率为350 W,超声功率为300 W,超声程序为运行1s后停止0~2s,如此循环,然后过滤;将滤渣按上述步骤重复提取2次,合并滤液,于3500 r/min条件下离心15 min,取上清液,在60℃浓缩至浸膏状,冷冻干燥后得莲子心总黄酮粗品;
3)将步骤2)将制得的莲子心总黄酮粗品投入超临界萃取釜,将质量浓度为95%的乙醇溶液和纯度大于99.99%的液态CO2按体积百分数之比为5:95作为萃取剂通入萃取釜进行分离后,将所得萃取液真空浓缩即得莲子心总黄酮。
实施例3
一种防治II型糖尿病的莲子心总黄酮的制备方法包括如下步骤:
1)称取新鲜莲子心,洗去杂质,50℃烘干至恒重后,超微粉碎至细度为15μm;
2)在步骤1)制备的莲子心粉末中按质量体积比1: 40(g/mL)加入质量浓度为80%的乙醇溶液,微波-超声波协同辅助提取5 min,微波功率为400 W,超声功率为400 W,超声程序为运行1s后停止0~2s,如此循环,然后过滤;将滤渣按上述步骤重复提取3次,合并滤液,于4000 r/min条件下离心10 min,取上清液,在65℃浓缩至浸膏状,冷冻干燥后得莲子心总黄酮粗品;
3)将步骤2)将制得的莲子心总黄酮粗品投入超临界萃取釜,将质量浓度为95%的乙醇溶液和纯度大于99.99%的液态CO2按体积百分数之比为10:90作为萃取剂通入萃取釜进行分离后,将所得萃取液真空浓缩即得莲子心总黄酮。
采用健康成年雄性昆明种小鼠建立高血糖模型进行降血糖效果试验。动物适应性饲养一周后无不良反应,开始试验:造型前随机取10只小鼠作为正常组,并禁食3h测血糖,作为基础血糖。然后将小鼠禁食不禁水15h后,尾静脉注射链脲佐菌素(STZ)100 mg/kg,7天后禁食3h筛选血糖值大于11.1 mmol/L的小鼠30只,按血糖水平随机分为1个阳性药物对照组、1个高血糖模型组和1个莲子心总黄酮实验组,每组10只,莲子心总黄酮实验组按100mg/(kg·d)进行灌胃给药,阳性药物对照组按100mg/(kg·d)给予盐酸二甲双弧,高血糖模型组给予等剂量生理盐水,连续灌胃21天,分别在给药第7、14及21天时将各组小鼠禁食12h,断尾取血测定各组小鼠空腹血糖值,记录数据并观察期间各组小鼠血糖变化情况,观察本发明莲子心总黄酮对高血糖模型小鼠空腹血糖的影响。在给药第25天时,各组小鼠禁食5h给药,并在给药20 min后,经口给予2.0 g/kg BW葡萄糖,尾静脉采血测定各组小鼠在0、0.5及2 h的血糖值,观察各组小鼠血糖变化情况,根据统计学方法求得耐糖量。
1. 本发明莲子心总黄酮对小鼠血糖的影响结果见表1。
表1 不同处理对小鼠血糖的影响(n=10)
注:A代表与正常组相比,差异极显著(p<0.01);a代表与正常组相比,差异显著(p<0.05);B代表与模型组相比,差异极显著(p<0.01);b代表与模型组相比,差异显著(p<0.05);D 代表与阳性对照组相比,差异极显著(p<0.01);d代表与阳性对照组相比,差异显著(p<0.05)。下同。
由表1可见,采用盐酸二甲双胍缓释片的阳性对照组小鼠的血糖下降百分率为42.89%,而采用本发明莲子心总黄酮的实验组小鼠血糖下降百分率达43.02%,说明本发明莲子心总黄酮具有显著的降血糖作用。
2. 本发明莲子心总黄酮的耐糖量指标的影响结果见表2。
表2 不同处理后小鼠的耐糖量指标(n=10)
由表2可见,与模型组相比,阳性对照组与实验组的耐糖量都降低,表明本发明莲子心总黄酮对糖耐量指标的影响结果呈阳性。
以上所述仅为本发明的较佳实施例,凡依本发明申请专利范围所做的均等变化与修饰,皆应属本发明的涵盖范围。
Claims (2)
1.一种防治II型糖尿病的莲子心总黄酮的制备方法,其特征在于:包括如下步骤:
1)称取新鲜莲子心,洗去杂质,40-50℃烘干至恒重后,超微粉碎至细度10-15 μm;
2)向步骤1)制备的莲子心粉末中加入乙醇溶液,微波-超声波协同辅助提取5-15 min,过滤;将滤渣按上述步骤重复提取2-3次,合并滤液,于3000-4000 r/min条件下离心10-20min,取上清液,在50-65℃浓缩至浸膏状,冷冻干燥后得莲子心总黄酮粗品;
3)将步骤2)制得的莲子心总黄酮粗品投入超临界萃取釜,将质量浓度为95%的乙醇溶液和液态CO2按体积百分数之比为1-10:90-99作为萃取剂通入萃取釜进行分离后,将所得萃取液真空浓缩即得莲子心总黄酮;
步骤2)中所用莲子心粉末重量与乙醇溶液的体积之比为1:20-40,所述乙醇溶液的质量浓度为70%-80%;
所述微波-超声波协同辅助提取中微波功率为300-400 W,超声功率为200-400 W,超声程序为运行1s后停止0~2s,如此循环。
2.根据权利要求1所述防治II型糖尿病的莲子心总黄酮的制备方法,其特征在于:步骤3)所述液态CO2的纯度大于99.99%。
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