CN113384640A - 一种葡萄叶提取物及其制备方法 - Google Patents
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Abstract
本发明公开了一种葡萄叶提取物及其制备方法，其以红色葡萄叶为原料提取精制而成的棕色至红色粉末提取物，具有该品种葡萄叶所特有的风味，在水及不同体积浓度乙醇、甲醇中能完全溶解；所述提取物的总多酚含量≥14％，花色苷含量≥0.3％，白藜芦醇含量≥0.005％。
Description
技术领域
本发明涉及植物提取技术领域，具体涉及一种葡萄叶提取物及其制备方法。
背景技术
葡萄是全球栽培最广的水果之一。国际葡萄与葡萄酒组织(OIV)最新报告显示，2018年全球葡萄栽培面积约为744万公顷，中国葡萄栽培面积约87.5万公顷。葡萄栽培过程伴随着大量副产物的产生，一个生长周期修剪废弃物量约达1～7.5吨/公顷，主要有葡萄叶、卷须、茎等，此类副产物多被丢弃、焚烧或堆肥，这些处理方式既浪费资源，也污染环境。葡萄叶在土耳其、希腊及中东一带大都作为烹饪材料，在美国和阿拉伯一些国家的饮食中，葡萄叶是一种不可或缺的辅料，与中国的粽子叶作用相似，巴勒斯坦西岸当地居民同时食用葡萄浆果和葡萄叶，由此可见，葡萄叶具有可食用的潜力。
现代学者研究已经发现葡萄叶中含有多酚、蛋白质、有机酸、皂苷、黄酮、白藜芦醇等化合物，其中多酚类化合物具有较强的抗氧化性、抗肿瘤、抗辐射、抑菌、免疫调节、保护心血管等作用；白藜芦醇是一种自由基清除剂和强效抗氧化剂，具有抗癌、心脏保护、神经保护和抗氧化等功能。因此，葡萄叶作为一种可利用的新资源，开发其利用价值是未来健康市场的大势所趋。目前国内现有的文献对葡萄籽、葡萄皮的研究较多，对葡萄叶研究寥寥无几，且并未对葡萄叶中的多个有效成分化合物进行同时提取富集，制备形成可广泛用于医药、食品及化妆品等领域的天然植物原料，这也导致葡萄叶资源没有得到充分利用而造成浪费。
因此，研究葡萄叶提取物制备方法，制备出品质较好的葡萄叶提取物，不仅可将葡萄叶资源变废为宝，减少环境污染，而且可为新资源的利用开辟新途径，具有非常广阔的应用前景。
发明内容
本发明的目的是克服了现有技术中的不足，提供了一种葡萄叶提取物的制备方法，该方法能得到含有多个有效成分化合物、品质较好的提取物，并适合工业化大生产，使葡萄叶的利用产业化、规模化，同时减少葡萄叶对环境的污染，增加葡萄种植户收入。
本发明提供的技术方案如下：
一种葡萄叶提取物的制备方法，具体步骤如下：
(1)采收、去除杂质，干燥：将葡萄叶中的杂质挑选出来，再进行自然阴干或低温烘干。
(2)浸泡：加入葡萄叶质量10～25倍的体积分数0.1～40％的乙醇浸泡一定的时间。
(3)提取：将上述浸泡液进行提取，提取温度为50～80℃，提取时间为1～8小时，提取次数为2次。
(4)过滤：将上述提取液过滤，收集滤液。
(5)浓缩：将上述滤液进行浓缩，浓缩温度为40～60℃，单次料液浓缩时间不超过6小时，浓缩液固含量不超过60％。
(6)干燥：将上述浓缩液进行冷冻干燥、低温干燥或瞬时高温干燥，低温干燥温度不超过60℃，瞬时高温干燥温度90～120℃。
(7)包装及储存：将上述干燥品用用薄膜袋包装，置阴凉干燥处避光储存。
优选的，步骤(1)中选用红色葡萄叶，具有该颜色葡萄叶的葡萄品种来源于烟七三、烟七四及蓝宝石中的一种或多种。
本发明还提供了采用上述方法制备得到的葡萄叶提取物。
优选的，葡萄叶提取物中总多酚含量≥14％，花色苷含量≥0.3％，白藜芦醇含量≥0.005％。
本发明与现有技术相比，具有以下有益效果：
本发明充分利用了废弃资源葡萄叶，将其制备成了同时富含较高含量的总多酚、花色苷、白藜芦醇的提取物，该种提取物可作为天然植物原料被广泛于医药、食品、化妆品等领域，开辟了天然植物原料新途径。
本发明制备方法工艺简单易操作，成本低廉，可适用于工业化，使葡萄叶的利用产业化、规模化，增加葡萄种植农户的收入，具有良好的经济效益。
具体实施方式
以下通过具体实施例对本发明技术方案做进一步解释说明。
以下具体实施方式中总多酚、花色苷、白藜芦醇的检测方法如下：
(1)总多酚的检测：准确称取焦性没食子酸标准品25mg至50mL容量瓶中，加入纯净水溶解并定容，分别取1mL、2mL、4mL、6mL、8mL、10mL、12mL、14mL标准品溶液至100mL容量瓶中，用纯净水定容，各吸取1mL，加入0.2mL福林酚和10mL15％碳酸钠溶液，充分混合，准时2min后，以水为对照，用紫外分光光度计在715nm处测定吸光度，绘制标准曲线。准确称取葡萄叶提取物30mg(M)至100mL锥形瓶中，加入30mL纯净水，沸水浴中回流加热30min，取出冷却至室温后，转移至50mL(V)棕色容量瓶中，并用纯净水定容，量取1mL样品试液，加入0.2mL福林酚和10mL15％碳酸钠溶液，充分混合，准时2min后，以水为对照，在715nm处测定吸光度，根据标准曲线读取多酚浓度C。总多酚含量(％)＝4.2×11.2×C×V/M×100。
(2)花色苷的检测：准确称取30mg(E)葡萄叶提取物至20mL容量瓶中，加入适量体积分数0.1％盐酸-甲醇试液，超声使溶解，用0.1％盐酸-甲醇试液定容，量取2mL至10mL容量瓶中，用0.1％盐酸-甲醇试液定容，用紫外分光光度计在528nm处测定吸光度A。花色苷含量(％)＝A×100/(772×E)。
(3)白藜芦醇的检测：以十八烷基硅烷键合硅胶为填充剂；以0.1％磷酸-乙腈(4：1)为流动相；检测波长为306nm；柱温30℃；流速1.0mL/min，进样量20uL。精密称定白藜芦醇对照品6mg(M1)置10mL容量瓶中，加50％乙腈使溶解并定容，精密量取本液5mL，加入50％乙腈至100mL，作为标准液。取本品1g(E)，精密称定，放置圆底烧瓶里，加入30mL的水使溶解。再加入30mL乙酸乙酯，强力搅拌5分钟。移置分液漏斗，分离乙酸乙酯层，在水层加入乙酸乙酯30mL，再两次强力搅拌5分钟，分离乙酸乙酯层，合并所有乙酸乙酯层，水浴挥干溶剂，在残留物加入甲醇5mL，再进行溶剂挥干。最终残留物用50％乙腈溶解转移至容量瓶中并定容至5mL，过滤，作为供试液。各取供试液和标准液20μL，在如上条件下按液相检测法进行检测，求供试液和标准液的峰面积AT和AS。白藜芦醇量
白藜芦醇含量
实施例1
一种葡萄叶提取物的制备方法，具体步骤如下：
(1)采收、去除杂质，干燥：选取蓝宝石品种的葡萄叶，在霜降后第1天采收，挑选出非葡萄叶类杂质，自然阴干。
(2)浸泡：称取500g葡萄叶，加入6000mL纯净水，浸泡1个小时。
(3)提取：将上述药液进行提取，提取温度为60℃，提取时间为8小时，提取次数为2次。
(4)过滤：将上述药液用滤布过滤，收集滤液。
(5)浓缩：将上述滤液进行浓缩，浓缩温度为50℃，单次料液浓缩时间为4小时，浓缩液固含量50％。
(6)干燥：将上述浓缩液进行低温真空干燥，干燥温度设定为60℃，得到固体138.7。
(7)包装及储存：将上述干燥品用用薄膜袋包装，置阴凉干燥处避光储存，即为成品葡萄叶提取物。产品得率27.7％，产品总多酚含量15.27％，花色苷含量0.28％，白藜芦醇含量0.0092％。
实施例2
一种葡萄叶提取物的制备方法，具体步骤如下：
(1)采收、去除杂质，干燥：选取烟七三品种的葡萄叶，在霜降后第2天采收，挑选出非葡萄叶类杂质，自然阴干。
(2)浸泡：称取500g葡萄叶，加入10000mL体积分数40％的乙醇，浸泡1个小时。
(3)提取：将上述药液进行提取，提取温度为70℃，提取时间为2小时，提取次数为2次。
(4)过滤：将上述药液用滤布过滤，收集滤液。
(5)浓缩：将上述滤液进行浓缩，浓缩温度为50℃，单次料液浓缩时间为4小时，浓缩液固含量40％。
(6)干燥：将上述浓缩液进行冷冻干燥，冷冻温度设定为-60℃，得到固体140.0g。
包装及储存：将上述干燥品用用薄膜袋包装，置阴凉干燥处避光储存，即为成品葡萄叶提取物。产品得率28.0％，产品总多酚含量23.3％，花色苷含量0.67％，白藜芦醇含量0.0093％。
实施例3
一种葡萄叶提取物的制备方法，具体步骤如下：
(1)采收、去除杂质，干燥：选取烟七三品种的葡萄叶，在霜降后第2天采收，挑选出非葡萄叶类杂质，自然阴干。
(2)浸泡：称取500g葡萄叶，加入10000mL纯净水，浸泡1个小时。
(3)提取：将上述药液进行提取，提取温度为70℃，提取时间为1小时，提取次数为2次。
(4)过滤：将上述药液用滤布过滤，收集滤液。
(5)浓缩：将上述滤液进行浓缩，浓缩温度为60℃，单次料液浓缩时间为4小时，浓缩液固含量45％。
(6)干燥：将上述浓缩液进行低温真空干燥，温度设定为60℃，得到固体157.5g。
包装及储存：将上述干燥品用用薄膜袋包装，置阴凉干燥处避光储存，即为成品葡萄叶提取物。产品得率31.5％，产品总多酚含量16.0％，花色苷含量0.36％，白藜芦醇含量0.0073％。
对比例1
一种葡萄叶提取物的制备方法，具体步骤如下：
(1)采收、去除杂质，干燥：选取山葡萄品种的葡萄叶，在霜降后2天采收，挑选出非葡萄叶类杂质，自然阴干。
(2)浸泡：称取500g葡萄叶，加入6000mL纯净水，浸泡1个小时。
(3)提取：将上述药液进行提取，提取温度为60℃，提取时间为8小时，提取次数为2次。
(4)过滤：将上述药液用滤布过滤，收集滤液。
(5)浓缩：将上述滤液进行浓缩，浓缩温度为50℃，单次料液浓缩时间为4小时，浓缩液固含量50％。
(6)干燥：将上述浓缩液进行低温真空干燥，温度设定为60℃，得到固体127.5g。
(7)包装及储存：将上述干燥品用用薄膜袋包装，置阴凉干燥处避光储存，即为成品葡萄叶提取物。产品得率25.5％，产品总多酚含量86.0％，花色苷含量0.03％，白藜芦醇含量0.0011％。
本对比例采用了山葡萄品种的葡萄叶制备提取物，与实施例1在提取条件完全相同的情况下，得到的提取物多酚含量虽然较高，但是花色苷及白藜芦醇含量非常低，表明结果不符合。
前述对本发明的具体示例性实施方案的描述是为了说明和例证的目的。这些描述并非想将本发明限定为所公开的精确形式，并且很显然，根据上述教导，可以进行很多改变和变化。对示例性实施例进行选择和描述的目的在于解释本发明的特定原理及其实际应用，从而使得本领域的技术人员能够实现并利用本发明的各种不同的示例性实施方案以及各种不同的选择和改变。本发明的范围意在由权利要求书及其等同形式所限定。
Claims (4)
1.一种葡萄叶提取物的制备方法，其特征在于，具体步骤如下：
(1)采收、去除杂质，干燥：将葡萄叶中的杂质挑选出来，再进行自然阴干或低温烘干。
(2)浸泡：加入葡萄叶质量10～25倍的体积分数0.1～40％的乙醇浸泡一定的时间。
(3)提取：将上述浸泡液进行提取，提取温度为50～80℃，提取时间为1～8小时，提取次数为2次。
(4)过滤：将上述提取液过滤，收集滤液。
(5)浓缩：将上述滤液进行浓缩，浓缩温度为40～60℃，单次料液浓缩时间不超过6小时，浓缩液固含量不超过60％。
(6)干燥：将上述浓缩液进行冷冻干燥、低温干燥或瞬时高温干燥，低温干燥温度不超过60℃，瞬时高温干燥温度90～120℃。
(7)包装及储存：将上述干燥品用用薄膜袋包装，置阴凉干燥处避光储存。
2.根据权利要求1所述的方法，其特征在于，步骤(1)中选用红色葡萄叶，具有该颜色葡萄叶的葡萄品种来源于烟七三、烟七四及蓝宝石中的一种或多种。
3.根据权利要求1或2所述方法制备得到的葡萄叶提取物。
4.根据权利要求3所述的葡萄叶提取物，其特征在于，葡萄叶提取物中总多酚含量≥14％，花色苷含量≥0.3％，白藜芦醇含量≥0.005％。
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