CN110693025A - 一种多功能成分协同降血糖的组合物及其应用 - Google Patents
一种多功能成分协同降血糖的组合物及其应用 Download PDFInfo
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Abstract
一种多功能成分协同降血糖的组合物及其应用,属于特殊用途食品技术领域。本发明针对单一天然活性成分降血糖效果有限的问题,将老山芹中黄酮、‘鲁赫’刺蔷薇果中多糖、马鹿茸多肽多种具有降血糖功能的成分配伍,发挥其协同增效作用,达到更高效的降血糖效果,是一款来源于森林资源的天然食品成分、具有高活性降血糖效果的组合物,可作为降血糖的辅助药物、营养增补剂、营养添加剂和功能因子应用于特膳食品和功能食品。
Description
技术领域
本发明属于特殊用途食品技术领域,具体涉及一种多功能成分协同降血糖的组合物及其应用。
背景技术
糖尿病是一种危害人类健康的常见病和多发病,随着人类生存环境和生活方式的改变,糖尿病患者的数量在世界范围内急剧增长,糖尿病已成为继心血管病、肿瘤之后的第三大病种,成为危害21世纪人类健康的杀手。如何防治糖尿病是世界范围内的重大研究课题,研发治疗糖尿病的药物受到人类高度重视。目前临床治疗糖尿病主要依靠注射各种胰岛素和口服降糖药物,但其会产生很多副作用,从天然产物中筛选降血糖功能成分就变得十分重要。黄酮、多糖和多肽具有显著的降血糖功能,从森林资源中提取制备高效、毒副作用小的降糖成分用于食品及功能性食品己成为研究热点。将黄酮、多糖、多肽三者进行科学合理复配,制成降血糖组合物尚未见报道。
老山芹又名东北牛防风、山芹菜,为伞形科独活属多年生草本植物,是一种具有食用和药用保健价值的春季山林野生蔬菜,在山野菜品种中占重要地位。其嫩茎叶口味鲜美,富含胡萝卜素、维生素A、维生素C、维生素B2、维生素E、铁、钙、蛋白质和多种氨基酸等营养成分及黄酮、香豆素、皂苷类化合物;老山芹全株可入药,味甘辛,性温,无毒,“益脾胃,去冷气,温中暖胃”,具有退热解毒、清洁血液、降低血糖、降压的功效。老山芹的功能性成分包括膳食纤维、黄酮和香豆素类化合物等(赵玉红,2018)。老山芹中黄酮含量很高,可作为黄酮提取源(刁绍起,2010)。有关老山芹中降血糖成分提取方面研究已有报道,但对其中黄酮及其与其他成分协同在降血糖产品中的应用尚未见报道。
植物多糖(plant polysaccharide),又称植物多聚糖,来源于植物的根、茎、叶、皮、种子和花,从各种中草药和其他植物中都可以提取分离出多糖,植物多糖是由许多相同或不同单糖以α-或β-糖苷键连接而成的多聚物化合物,聚合度超过10个,其分子量一般为数万甚至数百万,是构成生命的四大基本物质之一。
随着植物多糖的分离纯化技术的不断完善,多糖结构分析、理化特性、免疫学及药理学的研究不断深入,将植物多糖应用于食品和医药领域成为关注的焦点。‘鲁赫’刺蔷薇是由野生型刺蔷薇培育出来的新型品种,抗寒性和耐盐碱性较野生刺蔷薇有所提高(赵梦雅,2019)。‘鲁赫’刺蔷薇作为研发的新品种,目前对它活性成分及其功能性研究方面报道很少。‘鲁赫’刺蔷薇果中含有多种活性成分如花青素、多糖、总多肽、总黄酮、原花青素等。
鹿茸是雄鹿未骨化密生茸毛的幼角,主要含有氨基酸、总磷脂、脂肪酸、糖类、激素样物质、硫酸软骨素、多胺、肽类、维生素、酶类以及碱基类物质等,其中氨基酸成分占总成分的一半以上。鹿茸中的蛋白质主要是以角蛋白、胶原蛋白、胰岛素样生长因子和表皮成长因子等形式存在,具有提高机体的免疫功能和抗氧化能力,能够促进组织创伤的愈合,降血糖等生理作用。
现有技术所采用的能起到降血糖作用的原料主要是直接采用某些具有降血糖效果的食品原料,有些技术采用原料提取物进行降血糖研究并开发相关降糖产品,但只是对原料中的成分进行利用,缺少对不同原料中不同功能因子的协同利用,即利用功能成分相互作用后达到协同效果进行产品开发。
发明内容
针对单一天然活性成分降血糖效果有限的问题,本发明提供了一种多功能成分协同降血糖的组合物,所述组合物由以下重量份的原料组成:老山芹黄酮20-120份,鲁赫刺蔷薇果多糖10-60份和马鹿茸多肽5-30份。
优选地,所述多功能成分协同降血糖的组合物由以下重量份的原料组成:老山芹黄酮60份,鲁赫刺蔷薇果多糖30份和马鹿茸多肽15份。
进一步地限定,所述老山芹黄酮通过如下方法制备获得:
1)将老山芹烘干后粉碎过筛,获得老山芹粉末;
2)将老山芹粉末脱脂后,按照1g:(10-50)mL的料液比,将脱脂后的老山芹粉末与体积浓度为10%-100%的乙醇溶液混合,20-80℃,提取1-5h,得到提取液;
3)提取液离心后,得到的上清液经乙酸乙酯萃取,萃取液浓缩后真空冻干,经大孔吸附树脂初次纯化,再经聚酰胺柱二次纯化,得到老山芹黄酮。
进一步地限定,所述鲁赫刺蔷薇果多糖通过如下方法制备获得:
1)鲁赫刺蔷薇果脱脂后,按照1g:(10-50)mL的料液比,将脱脂后鲁赫刺蔷薇果与水混合,80-95℃提取3-5h,得到多糖粗提取液;
2)然后依次经脱蛋白、脱色后透析,所得透析液冻干后,经树脂层析分离,得到鲁赫刺蔷薇果多糖。
进一步地限定,所述马鹿茸多肽通过如下方法制备获得:
1)马鹿茸经去皮、粉碎后脱脂,用蒸馏水调节马鹿茸粉末质量浓度为5%-10%,利用Flavourzyme蛋白酶,pH6.5-7条件下,45-50℃酶解1h-3h,得到马鹿茸多肽粗提物,将马鹿茸多肽粗提物经超滤,收集分子量<10KDa的小分子多肽;以马鹿茸质量计,所述Flavourzyme蛋白酶加酶量为5000-6000U/g;
2)采用葡聚糖凝胶对超滤后的小分子多肽进行分离纯化,得到马鹿茸多肽。
进一步地限定,所述老山芹黄酮制备方法中步骤1)所述老山芹粉碎后过40-100目筛。
进一步地限定,所述鲁赫刺蔷薇果多糖制备方法中步骤2)所述脱蛋白采用Sevage法;所述脱色采用双氧水进行。
进一步地限定,所述鲁赫刺蔷薇果多糖制备方法中步骤2)所述树脂为DEAE树脂。
进一步地限定,所述马鹿茸多肽制备方法中步骤2)所述采用的葡聚糖凝胶为Sephadex LH-20。
本发明还提供了上述的组合物在制备预防或辅助治疗糖尿病的保健食品或药物中的应用。
有益效果
本发明将老山芹中黄酮、‘鲁赫’刺蔷薇果中多糖、马鹿茸多肽等多种具有降血糖功能的成分配伍,发挥其协同增效作用,经过体内和体外实验验证具有更高效的降血糖功能。是一款来源于林下资源天然食品成分、具有高活性降血糖效果的组合物,产品原料天然,对身体无伤害,可长期食用。同时制作过程中利用‘鲁赫’刺蔷薇果中多糖的流变学特性,在制作产品时加入,既能起到提高降血糖功能,又能起到粘合其他成分的作用,使产品制作时利用天然成分的特性,减少相关食品添加剂的使用,加工制备工艺更简易。
本发明从不同原料(老山芹、‘鲁赫’刺蔷薇、马鹿茸)中提取具有降血糖功能的成分,并将各成分进行配伍、组合,达到协同增效的作用,可作为降血糖的辅助药物、营养增补剂、营养添加剂和作为功能因子应用于特膳食品和功能食品,代表未来功能性食品及功能性食品基料的发展趋势,可以解决单一组分降血糖功能性弱的问题。
附图说明
图1老山芹黄酮高效液相色谱,横坐标为出峰时间(min),纵坐标为电信号,其中1为金丝桃苷;2为山奈酚;3为槲皮素;
图2多糖浓度对α-葡萄糖苷酶抑制率的影响;横坐标为鲁赫刺蔷薇果多糖浓度(ug/mL),纵坐标为抑制率(%);
图3多糖浓度对α-淀粉酶抑制率的影响:横坐标为鲁赫刺蔷薇果多糖浓度(ug/mL),纵坐标为抑制率(%);
图4不同浓度PⅠ、PⅡ、PⅢ和PⅣ的葡萄糖摄取率,横坐标中control为空白对照,insulin为胰岛素组,Metformin为二甲双胍(阳性对照组),纵坐标为葡萄糖摄取率;
图5不同浓度PⅠ、PⅡ、PⅢ和PⅣ对STZ损伤的胰岛β细胞修复的影响,横坐标中control为空白对照组,model为模型组,exendin-4为醋酸艾塞那肽(阳性对照组),纵坐标为活细胞比例。
具体实施方式
本发明将老山芹中黄酮、‘鲁赫’刺蔷薇果中多糖、马鹿茸多肽等多种具有降血糖功能的成分配伍,发挥其协同增效作用,达到更高效的降血糖效果,研制一款来源于天然食品成分、具有高活性降血糖效果的组合物。
技术上通过证明每种成分具有降血糖功能,将不同功效成分配伍,证明其协同增效作用,开发具有更高降血糖效果的产品。
下面具体描述本发明制备的多功能成分协同降血糖的组合物,所用药品,试剂以及仪器设备,如无特殊说明,均可通过商业化途径购买获得。
实施例1.多功能成分协同降血糖的组合物。
本实施例所述的多功能成分协同降血糖的组合物由以下重量份的原料组成:老山芹黄酮60份,鲁赫刺蔷薇果多糖30份和马鹿茸多肽15份。
通过如下的方法制备获得:
一、老山芹黄酮的制备:
1)老山芹经清洗、烘干,用粉碎机充分粉碎后,过60目筛;
2)过筛后采用超临界CO2萃取方法,在萃取时间1.5h、萃取温度40℃、萃取压力35MPa下脱脂,将脱脂后老山芹,按照1g:30mL的料液比与体积浓度为50%的乙醇溶液混合,60℃,提取3h,得到提取液;
3)提取液经离心后,在旋转蒸发仪中浓缩;提取物用乙酸乙酯萃取,将萃取物浓缩液真空冻干,得老山芹黄酮提取物。提取物经聚酰胺柱纯化,上样质量浓度为1.80mg/mL,上样液50mL上聚酰胺柱,用4~5BV蒸馏水冲洗,然后分别用20%-90%体积浓度的乙醇溶液100mL以1mL/min进行分步分段洗脱,乙醇溶液浓度梯度分别为20%、30%、40%、50%、60%、70%、80%、90%,收集50%和70%流出液,得到的黄酮纯度为72.15%。经高效液相色谱分析检测老山芹黄酮中活性物质含量,如图1所示,其中金丝桃苷含量为0.62%,山奈酚为3.39%,槲皮素为0.66%。
表1老山芹各活性物质含量
二、鲁赫刺蔷薇果多糖制备:
1)鲁赫刺蔷薇果利用超临界CO2萃取方法,在萃取时间2h、萃取温度35℃、萃取压力40MPa下脱脂,按照1g:30mL的料液比,将鲁赫刺蔷薇果与水混合,90℃提取3h,得到多糖粗提取液;
2)得到的多糖液用Sevage法脱蛋白(Sevage试剂中氯仿与正丁醇的比例为4:1、试剂加入量为多糖溶液体积的1/3、震摇时间为25min、脱蛋白1次),双氧水脱色(在H2O2体积分数为6%、脱色温度为50℃、pH值为7、脱色时间为90min)装入透析袋中在蒸馏水中透析24h,旋转蒸发仪浓缩得粗多糖。粗多糖加入少量蒸馏水中溶解,样品溶液离心(5000r/min,10min)以除去不溶物,然后将离心液加样到DEAE Sepharose色谱柱(20mm×300mm,10cm),分别用蒸馏水及0.1、0.2mol/L和0.5mol/L NaCl溶液洗脱,时长6h,利用试管收集洗脱液(5mL/管,1mL/min),透析除去NaCl,冻干,得到鲁赫刺蔷薇果多糖。
采用苯酚-硫酸比色法测定本发明提取获得的鲁赫刺蔷薇果多糖成分及含量如下:
表2鲁赫’刺蔷薇果多糖成分分析
三、马鹿茸多肽制备:
1)将新鲜的马鹿茸去除皮毛,切成小块,用去离子水冲洗至无血色,再用冷冻干燥机进行干燥,低温粉碎,过80目筛,石油醚去脂,于-40℃下保存备用。取去脂后马鹿茸粉,加蒸馏水配制成底物浓度10%,加Flavourzyme蛋白酶,在50℃温度下进行酶解,反应1h后置90℃水浴中灭酶10min,4000r/min离心20min,取酶解上清液,用超滤离心管(3kDa)进行超滤离心,用去离子水将样品质量浓度配制成5%,转速4000rpm,离心40min,收集管底溶液冻干制粉用于下一步分离纯化实验,回收管芯内溶液冻干制粉,-40℃保存。
2)将Sephadex LH-20葡聚糖凝胶浸泡在体积分数为60%的乙醇中,不断搅拌使之充分溶胀,浸泡24h后,用去离子水洗去残留的乙醇直至无醇味,最后将其装入规格为50cm×3.5cm的层析柱。用去离子水溶解小分子肽,配制成55mg/mL的溶液,加样量为1.5mL、加样浓度为55mg/mL、洗脱流速为0.25mL/min,在此条件下可以得到四个组分,分别为PⅠ、PⅡ、PⅢ、PⅣ,取PⅢ为本发明所使用的马鹿茸多肽。
四、将上述得到的老山芹黄酮、鲁赫刺蔷薇果多糖和马鹿茸多肽按重量份充分混匀,即得所述的组合物。
考察本发明制备的组合物的降血糖的功效。
本实施例首先分别考察了本发明方法提取的老山芹黄酮、鲁赫刺蔷薇果多糖和马鹿茸多肽单独使用时降糖效果,具体如下:
1.老山芹黄酮对HepG2细胞的葡萄糖消耗作用及细胞增殖作用。
人肝癌组织细胞系HepG2,由黑龙江省哈尔滨医科大学惠赠。
细胞复苏与传代方法:在离心管中加入4mL含有10%FBS的培养液,然后将HepG2细胞从液氮罐中迅速取出,放入37℃的温水的水浴锅中,使其融化至无冰碴。移入离心管中,吹打均匀后,离心机l000r/min离心5min,弃去上清液,加入4mL培养液,将细胞移入培养瓶中,放置到条件为37℃、5%CO2的二氧化碳培养箱中,按1:3比例进行传代。
MTT试验:选取生长对数期的细胞经0.25%胰蛋白酶进行消化后,用含10%新生牛血清DMEM培养液制成单细胞悬液,每孔按照5×104个细胞接种于96孔细胞板内,待细胞完全贴壁后,实验组以每孔100μL分别加入1000μg/mL、500μg/mL、100μg/mL、50μg/mL、10μg/mL、5μg/mL、1μg/mL含药的培养液,每组复6孔,于37℃,5%CO2培养箱培养24h后,按1:9体积每孔加入5g/L MTT溶液继续培养4h,弃去培养液,每孔加入150μL二甲基亚砜(DMSO),使结晶充分溶解,于酶标仪490nm处测定吸光值,以吸光值反应细胞活性和数量的多少。
以胰岛素、阿卡波为对照,采用Origin 9.2制图,采用SPSS 20.0对数据进行方差分析、显著性检验,显著性水平设置p<0.05。结果如下表所示。
表3老山芹中黄酮对HepG2细胞的葡萄糖消耗作用及细胞增殖作用
2.老山芹中黄酮对α-淀粉酶的抑制作用。
α-淀粉酶抑制率测定方法如下:取2mg/mL的待测样品100μL,加入50μL的α-淀粉酶溶液,于37℃水浴10min后,加入1.0%的可溶性淀粉溶液50μL,于37℃水浴5min后,立即加入DNS溶液100μL终止反应,再加入1000μL PBS,用酶标仪在波长为540nm处测定吸光值A1,另取100μL磷酸盐缓冲溶液代替酶解液,测定其吸光值A0,再测定只有酶解液反应体系额吸光值A2,每组试验做5次平行。计算α-淀粉酶抑制率,公式为:
α-淀粉酶抑制率(%)=[(A1-A0)/A2]×100
3.鲁赫刺蔷薇果多糖对α-葡萄糖苷酶以及α-淀粉酶的抑制作用。
α-葡萄糖苷酶抑制率测定方法如下:取2mg/mL的待测样品100μL,加入α-葡萄糖苷酶溶液50μL,于37℃水浴10min后,10mg/mL的底物PNPG溶液50μL,于37℃水浴15min后,立即加入1mol/L的Na2CO3溶液100μL终止反应,用酶标仪在波长为405nm处测定吸光值A1,另取100μL磷酸盐缓冲溶液代替酶解液,测定其吸光值A0,再测定只有酶解液反应体系额吸光值A2,每组试验做5次平行。计算α-葡萄糖苷酶抑制率,公式为:
α-葡萄糖苷酶抑制率(%)=[(A1-A0)/A2]×100。
α-淀粉酶抑制率测定方法如下:取2mg/mL的待测样品100μL,加入50μL的α-淀粉酶溶液,于37℃水浴10min后,加入1.0%的可溶性淀粉溶液50μL,于37℃水浴5min后,立即加入DNS溶液100μL终止反应,再加入1000μL PBS,用酶标仪在波长为540nm处测定吸光值A1,另取100μL磷酸盐缓冲溶液代替酶解液,测定其吸光值A0,再测定只有酶解液反应体系额吸光值A2,每组试验做5次平行。计算α-淀粉酶抑制率,公式同前。结果如图2及图3所示。
4.马鹿茸多肽降血糖效果
表4不同浓度马鹿茸降血糖肽纯化组分对HepG2细胞葡萄糖消耗作用及细胞增殖的影响
注:字母不同代表差异显著(P<0.05)
低浓度组分PⅢ(0.05mg/mL)的葡萄糖消耗量最高,其作用高于试验条件下胰岛素和二甲双胍的作用;低浓度(0.01mg/mL、0.05mg/mL)的四个组分均可使HepG2细胞的存活率保持良好,其中组分PⅢ(0.01mg/mL)和组分PⅣ(0.01mg/mL)细胞存活率均高于胰岛素和二甲双胍作用的细胞的存活率;高剂量的马鹿茸降血糖肽纯化组分对胰岛β细胞增殖较对照组有显著增加(P<0.05),其中高剂量的组分PⅢ对细胞的增殖作用接近于阳性对照Exendin-4作用的胰岛β细胞增殖作用;高剂量的组分PⅢ细胞增殖率高于其他组分,接近于阳性对照组的效果,如图4、5所示。
黄酮、多糖、多肽协同降糖效果考察:本发明设定了多组组合,考察不同活性组分的组合对α-淀粉酶抑制的协同作用,下述各组药物实验中各药物浓度有所不同,但使用的体积相同,结果如下表所示:
表5不同组分对α-淀粉酶抑制的协同作用
注:IR代表相应浓度时α-淀粉酶抑制率;Total IR代表三种组分α-淀粉酶抑制率的总和。
从表5可以看出三种化合物对α-淀粉酶抑制作用,显示出对α-淀粉酶抑制活性的强弱顺序是:多肽>黄酮>多糖,当多肽的浓度有0.05mg/mL时,其酶抑制率达到17.09%,当与黄酮混合配成样品时,其混合样的酶抑制率为78.67%,和它们单独的抑制率总和59.24%相比较,抑制率有明显的提高;多肽和黄酮配成的样品的酶抑制率比三者单独的酶抑制率总和也有明显的提髙,而多糖、多糖-黄酮及多糖-多肽的样品体系对酶的抑制率与相应各组分对酶抑制率的总和相比有小幅提升。三种混合物对酶的抑制率与它们抑制率总和相比,有显著的提高。
空腹血糖值是糖尿病最常用的检测指标,反映胰岛β细胞功能,一般表示基础胰岛素的分泌功能。
将小鼠随机分成10组,分别为黄酮组、多糖组、多肽组、黄酮+多糖组、黄酮+多肽组、多糖+多肽组、黄酮+多糖+多肽组、阳性对照组、模型组和空白对照组,每组10只。阳性对照组按照小鼠的体重灌胃剂量为200mg/kg二甲双胍溶液;模型组和空白组用生理盐水进行灌胃;黄酮、多糖、多肽分别为0.2mg/mL、0.1mg/mL、0.05mg/mL浓度,按100mg/kg的剂量灌胃。根据小鼠的体重灌胃体积为0.1mL/10g,每天灌胃一次,并观察饮水量、进食量变化,灌胃给药的第0、7、14、21、28d测空腹血糖值并记录,持续28d。28d内每隔7d对小鼠的空腹血糖进行测定,结果见表6。
表6受试物对小鼠血糖的影响
注:与空白对照组比,**表示差异极显著,p<0.01;*表示差异显著,p<0.05;与模型组相比,##表示差异极显著p<0.01;#表示差异显著,p<0.05;与阳性对照组比,aa表示差异极显著,p<0.01;a表示差异显著,p<0.05。
在持续灌胃的28d中,各组糖尿病小鼠血糖均极显著高于空白对照组(p<0.01),受试组小鼠血糖下降均较快,黄酮+多糖+多肽组效果强于阳性对照组,说明本发明制备的组合物降糖效果较好。
脂类在机体内的主要功能是氧化功能。正常情况下血清中脂类含量相对恒定,当机体患有糖尿病时,常见血脂异常,脂类代谢发生障碍,表现为TC、TG水平升高,HDL-C水平降低。具体结果如表7所示。
表7受试物对小鼠血脂的影响
注:与空白对照组比,**表示差异极显著,p<0.01;*表示差异显著,p<0.05;与模型组相比,##表示差异极显著p<0.01;#表示差异显著,p<0.05;与阳性对照组比,aa表示差异极显著,p<0.01;a表示差异显著,p<0.05。
由表7所示,各受试组和阳性对照组小鼠的TC、TG均显著低于模型组(p<0.05),HDL-C则显著升高(p<0.05),说明受试物和阳性药物对小鼠血脂的恢复,抑制小鼠脂代谢障碍有一定作用。
综上可见,本发明采用三种功能因子组合协同增效作用,产品具有更好的降血糖效果。
本发明还可以利用‘鲁赫’刺蔷薇果中多糖优良的流变学特性,制作压片,既能起到提高降血糖功能,又能起到粘结其他成分的作用,使压片制作不需添加硬脂酸镁等赋形剂,在减少食品添加剂的使用的同时又简化了加工制备工艺。
虽然本发明已以较佳的实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可以做各种改动和修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (10)
1.一种多功能成分协同降血糖的组合物,其特征在于,所述组合物由以下重量份的原料组成:老山芹黄酮20-120份,鲁赫刺蔷薇果多糖10-60份和马鹿茸多肽5-30份。
2.根据权利要求1所述的组合物,其特征在于,所述组合物由以下重量份的原料组成:老山芹黄酮60份,鲁赫刺蔷薇果多糖30份和马鹿茸多肽15份。
3.根据权利要求1所述的组合物,其特征在于,所述老山芹黄酮通过如下方法制备获得:
1)将老山芹烘干后粉碎过筛,获得老山芹粉末;
2)将老山芹粉末脱脂后,按照1g:(10-50)mL的料液比,将脱脂后的老山芹粉末与体积浓度为10%-100%的乙醇溶液混合,20-80℃,提取1-5h,得到提取液;
3)提取液离心后,得到的上清液经乙酸乙酯萃取,萃取液浓缩后真空冻干,经大孔吸附树脂初次纯化,再经聚酰胺柱二次纯化,得到老山芹黄酮。
4.根据权利要求1所述的组合物,其特征在于,所述鲁赫刺蔷薇果多糖通过如下方法制备获得:
1)鲁赫刺蔷薇果脱脂后,按照1g:(10-50)mL的料液比,将脱脂后鲁赫刺蔷薇果与水混合,80-95℃提取3-5h,得到多糖粗提取液;
2)然后依次经脱蛋白、脱色后透析,所得透析液冻干后,经树脂层析分离,得到鲁赫刺蔷薇果多糖。
5.根据权利要求1所述的组合物,其特征在于,所述马鹿茸多肽通过如下方法制备获得:
1)马鹿茸经去皮、粉碎后脱脂,用蒸馏水调节马鹿茸粉末质量浓度为5%-10%,利用Flavourzyme蛋白酶,pH6.5-7条件下,45-50℃酶解1h-3h,得到马鹿茸多肽粗提物,将马鹿茸多肽粗提物经超滤,收集分子量<10KDa的小分子多肽;以马鹿茸质量计,所述Flavourzyme蛋白酶加酶量为5000-6000U/g;
2)采用葡聚糖凝胶对超滤后的小分子多肽进行分离纯化,得到马鹿茸多肽。
6.根据权利要求3所述的组合物,其特征在于,步骤1)所述老山芹粉碎后过40-100目筛。
7.根据权利要求4所述的组合物,其特征在于,步骤2)所述脱蛋白采用Sevage法;所述脱色采用双氧水进行。
8.根据权利要求4所述的组合物,其特征在于,步骤2)所述树脂为DEAE树脂。
9.根据权利要求5所述的组合物,其特征在于,步骤2)所述采用葡聚糖凝胶为SephadexLH-20。
10.权利要求1-9任意一项所述的组合物在制备预防或辅助治疗糖尿病的保健食品或药物中的应用。
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