CN110754601B - 一种高抑菌性葡萄籽提取物的制备方法 - Google Patents
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Abstract
本发明涉及植物提取物改性技术领域,公开了一种高抑菌性葡萄籽提取物的制备方法,步骤为:将葡萄籽粉碎过筛得到葡萄籽粉末;向葡萄籽粉末中加入乙醇,超声提取得到葡萄籽提取液;将葡萄籽提取液真空干燥后得到葡萄籽提取物;用低温等离子体气氛处理葡萄籽提取物,得到高抑菌性葡萄籽提取物。本发明利用低温等离子体的高能粒子和活性组分与葡萄籽提取物表面发生作用,可以对葡萄籽提取物中的多酚类等成分表面进行修饰改性,从而有效提高了葡萄籽提取物的抑菌活性。
Description
技术领域
本发明涉及植物提取物改性技术领域,尤其是涉及一种高抑菌性葡萄籽提取物的制备方法。
背景技术
葡萄是世界上普遍栽培的水果之一,中国年产葡萄约140万t,其中约80%用于酿酒。葡萄籽是葡萄酒酿造工业的副产品,在葡萄酒酿造过程中,会产生葡萄总量3%的葡萄籽。
葡萄籽中含有丰富的多酚类物质,具有一定的抑菌性和抗氧化能力,因此富含多酚类物质的葡萄籽提取物已广泛应用于食品、药品、化妆品等领域进行抑菌。例如,在中国专利文献上公开的“一种利用复配植物多酚对生鲜调理猪肉的抑菌保鲜方法”,其公告号CN109042830A,该发明通过茶多酚和葡萄籽提取物对生鲜调理猪肉进行抑菌,能显著抑制生鲜调理猪肉在贮藏过程中热死环丝菌、假单胞菌、肠杆菌及葡萄球菌的生长,对挥发性氨基氮有显著地抑制作用,并且能够维持生鲜调理猪肉在贮藏过程中的pH、硬度及a值化,保持产品贮藏品质。
但葡萄籽提取物的抑菌性能还有待提高,做抑菌剂时需与其他组分复配抑菌效果才能达到使用要求,因此寻找可以有效提高葡萄籽提取物的方法有重要意义。
发明内容
本发明是为了克服现有技术中葡萄籽中含有丰富的多酚类物质,具有一定的抑菌性,可以广泛应用于食品、药品、化妆品等领域进行抑菌,但其抑菌性能还有待提高,需与其他组分复配抑菌效果才能达到使用要求的问题,提供一种高抑菌性葡萄籽提取物的制备方法,利用低温等离子体技术对葡萄籽提取物进行改性,有效提高了其抑菌活性。
为了实现上述目的,本发明采用以下技术方案:
一种高抑菌性葡萄籽提取物的制备方法,包括如下步骤:
(1)将葡萄籽粉碎过筛得到葡萄籽粉末;
(2)向葡萄籽粉末中加入乙醇,超声提取得到葡萄籽提取液;
(3)将葡萄籽提取液真空干燥后得到葡萄籽提取物;
(4)用低温等离子体气氛处理葡萄籽提取物,得到高抑菌性葡萄籽提取物。
本发明步骤(1)-(3)中以乙醇为溶剂,采用超声提取法提取得到葡萄籽提取物,乙醇对植物细胞的穿透能力也较强,且葡萄籽中的多酚类物质具有多元酚羟基结构和一定的极性,易溶于亲水性有机溶剂乙醇,因此多酚类物质可以被有效提取;而淀粉、蛋白质、果胶、黏液质等不溶于乙醇的物质则不能被提取,因此得到的葡萄籽提取物中主要成分为多酚类物质,具有抑菌性。
再通过步骤(4)中用低温等离子技术处理葡萄籽提取物。等离子体是物质在自然条件下的第四种状态,是气体分子受到辐射、热或者受到外加电场等能量激发而电离生成的包含离子、电子、中性原子(基态或激发态)、自由基等多种粒子的集合体,其在生成过程中会产生具有化学键破坏能力的电磁场、热、紫外线、带电粒子、激发态粒子、亚稳态粒子等活性组分,利用低温等离子体的高能粒子和活性组分与葡萄籽提取物表面发生作用,可以对葡萄籽提取物中的多酚类等成分表面进行修饰改性,如产生醌类等物质,从而提高葡萄籽提取物的抑菌活性。
作为优选,步骤(4)中产生低温等离子体气氛的工作气体为体积比为1:1~3:1的O2和N2。在此气氛下可以有效对葡萄籽提取物中的多酚类成分进行改性,生成醌类等物质,提高其抑菌活性。
作为优选,步骤(4)中处理功率为600~800W。
作为优选,步骤(4)中处理时间为10~15min。在此处理低温等离子体处理功率和时间下,得到的产物的抑菌性能最佳。
作为优选,步骤(1)中过60~100目筛。采用此粒径范围的葡萄籽粉末,有利于对葡萄籽粉末中多酚类成分的提取,从而提高葡萄籽提取物的抑菌性能。
作为优选,步骤(2)中加入乙醇的料液比为1:(7~9),乙醇的体积分数为40~60%。乙醇浓度过低不能有效提取葡萄籽粉末中的多酚类物质,而乙醇浓度过高,溶剂与多酚极性差异增大,醇溶性杂质等亲脂性成分溶出量增加,与多酚类物质竞争溶剂结合,导致多酚类物质的提取率下降,从而使得葡萄籽提取物的抑菌性能下降。
作为优选,步骤(2)中超声频率35~45Hz,超声时间40~50min,提取温度35~55℃。在此超声条件和温度下可以保证葡萄籽中的有效成分不被破坏并且可以被充分提取。
作为优选,步骤(3)中真空干燥温度60~80℃。
因此,本发明具有如下有益效果:以乙醇为溶剂,采用超声提取法得到的葡萄籽提取物中多酚类含量高,具有一定的抑菌性能,然后利用低温等离子体的高能粒子和活性组分与葡萄籽提取物表面发生作用,可以对葡萄籽提取物中的多酚类等成分表面进行修饰改性,如产生醌类等物质,从而有效提高了葡萄籽提取物的抑菌活性。
具体实施方式
下面结合具体实施方式对本发明做进一步的描述。
实施例1:
(1)将葡萄籽粉碎,过80目筛得到葡萄籽粉末;
(2)向葡萄籽粉末中加入体积分数为50%的乙醇,料液比为1:8,40℃,40Hz下超声提取45min得到葡萄籽提取液;
(3)将葡萄籽提取液70℃真空干燥后得到葡萄籽提取物;
(4)用低温等离子体气氛处理葡萄籽提取物得到高抑菌性葡萄籽提取物,处理功率700W,处理时间13min,产生低温等离子体气氛的工作气体为体积比为2:1的O2和N2。
实施例2:
(1)将葡萄籽粉碎,过60目筛得到葡萄籽粉末;
(2)向葡萄籽粉末中加入体积分数为40%的乙醇,料液比为1:7,35℃,45Hz下超声提取50min得到葡萄籽提取液;
(3)将葡萄籽提取液60℃真空干燥后得到葡萄籽提取物;
(4)用低温等离子体气氛处理葡萄籽提取物得到高抑菌性葡萄籽提取物,处理功率600W,处理时间15min,产生低温等离子体气氛的工作气体为体积比为1:1的O2和N2。
实施例3:
(1)将葡萄籽粉碎,过100目筛得到葡萄籽粉末;
(2)向葡萄籽粉末中加入体积分数为60%的乙醇,料液比为1:9,55℃,35Hz下超声提取40min得到葡萄籽提取液;
(3)将葡萄籽提取液80℃真空干燥后得到葡萄籽提取物;
(4)用低温等离子体气氛处理葡萄籽提取物得到高抑菌性葡萄籽提取物,处理功率800W,处理时间10min,产生低温等离子体气氛的工作气体为体积比为3:1的O2和N2。
对比例1:
对比例1与实施例1的区别在于,不用低温等离子体气氛处理葡萄籽提取物,其余均与实施例1中相同。
对比例2:
对比例2与实施例1的区别在于,用低温等离子体气氛处理葡萄籽提取物时,处理时间5min,其余均与实施例1中相同。
对比例3
对比例3与实施例1的区别在于,用低温等离子体气氛处理葡萄籽提取物时,产生低温等离子体气氛的工作气体为体积比为1:2的O2和N2,其余均与实施例1中相同。
对比例4:
对比例4与实施例1的区别在于,用体积分数为90%的乙醇进行提取,其余均与实施例1中相同。
对上述实施例和对比例中制得的葡萄籽提取物或高抑菌性葡萄籽提取物对大肠杆菌(Ec)、金黄色葡萄球菌(Sa)、沙门氏菌(Ty)、酿酒酵母(Sc)、青霉菌(Pe)的抑菌性能进行测试,结果如表1所示。
测试方法:
1、供试菌株的准备:在无菌室中将供试菌种接入相对应的试管斜面培养基上,细菌置于35℃恒温培养箱内培养24h;霉菌、酵母菌置于28℃恒温培养箱内培养45h后置于4℃冰箱冷藏备用。其中细菌培养使用牛肉膏蛋白胨培养基:牛肉膏0.3g,蛋白胨1.0g,NaCl0.5g,琼脂1.5g,水100mL,pH 7.2;酵母菌及霉菌培养使用豆芽汁葡萄糖培养基:黄豆芽10g,葡萄糖5g,琼脂1.5g,水100ml,自然pH。
2、菌悬浮液的准备:用接种环挑取少量菌体于装有9mL无菌水的试管中,制成菌悬浮液,用血球计数板计数,调整菌悬浮液的浓度至含孢子或菌体为106~107个/mL。
3、葡萄籽提取物溶液的制备:用蒸馏水将实施例和对比例中制得的高抑菌性葡萄籽提取物分别配制成浓度为0.1mg/mL、1mg/mL、10mg/mL、100mg/mL、1000mg/mL的溶液待用。
4、抑菌实验:取直径为6mm的灭菌滤纸片放入不同葡萄籽提取物溶液中浸泡过夜,将各种待试菌悬液各取0.5mL与相应固体培养基制成含菌平板,用无菌镊子把葡萄籽提取物溶液的滤纸片贴在含菌平板上,每皿3片,进行三次平行实验。细菌置于37℃恒温培养箱内培养24h,霉菌、酵母菌置于29℃恒温培养箱内培养48h,测量滤纸片的抑菌圈直径,比较各葡萄籽提取物溶液的抑菌性能。
表1:高抑菌性葡萄籽提取物抑菌性能测试结果(抑菌圈直径:mm)。
从表1中可以看出,实施例1-3中,使用本发明中的方法制备出的高抑菌性葡萄籽提取物对五种菌的抑菌圈直径比对比例1中没有经过低温等离子体气氛处理的葡萄籽提取物的抑菌圈直径提高了近两倍;且实施例1-3中在浓度为100mg/mL时对酿酒酵母和青霉菌已经产生抑菌圈;而对比例1中在浓度为1000mg/mL时才对酿酒酵母和青霉菌产生抑菌圈。
对比例2和3改变低温等离子体处理时的条件,使其落在本发明的范围外,得到的葡萄籽提取物的抑菌性能与实施例1相比有明显下降;对比例4中改变提取时的乙醇浓度,最终得到的葡萄籽提取物的抑菌性能与实施例1相比也有明显下降,证明本发明中的提取和处理条件不是常规选择。
Claims (7)
1.一种高抑菌性葡萄籽提取物的制备方法,其特征是,包括如下步骤:
(1)将葡萄籽粉碎过筛得到葡萄籽粉末;
(2)向葡萄籽粉末中加入乙醇,超声提取得到葡萄籽提取液;
(3)将葡萄籽提取液真空干燥后得到葡萄籽提取物;
(4)用低温等离子体气氛处理葡萄籽提取物,得到高抑菌性葡萄籽提取物;产生低温等离子体气氛的工作气体为体积比为1:1~3:1的O2和N2。
2.根据权利要求1所述的一种高抑菌性葡萄籽提取物的制备方法,其特征是,步骤(4)中处理功率为600~800W。
3.根据权利要求1所述的一种高抑菌性葡萄籽提取物的制备方法,其特征是,步骤(4)中处理时间为10~15min。
4.根据权利要求1所述的一种高抑菌性葡萄籽提取物的制备方法,其特征是,步骤(1)中过60~100目筛。
5.根据权利要求1所述的一种高抑菌性葡萄籽提取物的制备方法,其特征是,步骤(2)中加入乙醇的料液比为1:(7~9),乙醇的体积分数为40~60%。
6.根据权利要求1或5所述的一种高抑菌性葡萄籽提取物的制备方法,其特征是,步骤(2)中超声频率35~45Hz,超声时间40~50min,提取温度35~55℃。
7.根据权利要求1所述的一种高抑菌性葡萄籽提取物的制备方法,其特征是,步骤(3)中真空干燥温度60~80℃。
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