CN111602818A - 一种具有明目抗蓝光效果的枸杞和黑果枸杞组合物 - Google Patents
一种具有明目抗蓝光效果的枸杞和黑果枸杞组合物 Download PDFInfo
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Abstract
本发明提供了一种明目抗蓝光的枸杞和黑果枸杞的组合物。包括以下重量份数的各组分：枸杞提取物1份～9份、黑果枸杞1份～9份。采用枸杞为原料，通过亚临界提取技术制备了枸杞提取物。采用黑果枸杞为原料，通过高速剪切技术、膜浓缩技术及大孔树脂层析技术制备了黑果枸杞提取物。并将枸杞提取物和黑果枸杞提取物以合适的比例混合，制备出一种具有明目抗蓝光活性的枸杞和黑果枸杞的组合物，能够有效保护长期使用电子产品的人群的视力，预防眼科疾病的发生。
Description
技术领域
本发明属于保健食品技术领域，具体涉及一种具有明目抗蓝光效果的枸杞和黑果枸杞组合物。
背景技术
随着各种电子设备的普及，波长在400nm～500nm之间的高能短波光对人体的伤害不容忽视，主要表现在造成视疲劳、近视、白内障以及黄斑病变等病理伤害。蓝光的波长短，在成像时聚焦点落在离视网膜更靠前的位置，因此长时间在蓝光照射下，眼球会处于紧张状态，引起视疲劳，可能导致人们出现视力模糊、下降等症状，从而影响人们的正常学习与工作。
目前，市场上已有的缓解视疲劳、保护视力的保健食品所用原料主要以枸杞子、决明子、菊花、越橘提取物等为主。但这些产品在某一方面都存在着一定的缺陷。枸杞属茄科枸杞属植物，浆果中富含多种枸杞色素，研究表明，枸杞色素具有多种生理活性，如缓解视力疲劳、提高人体免疫功能、预防和抑制肿瘤及预防动脉粥样硬化等作用。枸杞虽然具有多种生理活性，但是总体上来说药效还是偏热，如果长期大量的服用，会引起“上火”的症状。决明子具有明目养肝的功效，但是同时也具有一定的降血压作用，会使很多身体虚弱或者血压过低的患者病情加重，身体会越来越差，甚至有生命危险。因此，就需要不断寻求新的保护视力且副作用小的产品。
发明内容
本发明的目的是提供一种明目抗蓝光的枸杞和黑果枸杞的组合物，以预防眼科疾病的发生。
本发明所提供的具有明目抗蓝光效果的枸杞和黑果枸杞的组合物，包含有枸杞提取物和黑果枸杞提取物；
其中所述枸杞提取物制备方法如下：
将枸杞粉碎，在枸杞粉末中加入萃取溶剂后置于亚临界萃取设备中进行萃取，然后收集脂溶性成分，灭菌，得到枸杞提取物。
其中萃取溶剂为食品添加剂丙烷或丁烷，
枸杞粉末与萃取溶剂质量体积比为1:10～1:40(g/ml)，萃取压力为1Mpa～10Mpa，萃取时间为2～5h，萃取温度为20℃～50℃；
所述黑果枸杞提取物的制备方法，是将黑果枸杞用水为提取溶剂进行高速剪切提取，离心后，获得黑果枸杞提取液；然后将黑果枸杞提取液先用10000～100000Da的膜去除大分子物质，再用孔径大小为160～360Da的膜进行浓缩；浓缩液采用大孔树脂层析柱吸附，用乙醇溶液洗脱，得到黑果枸杞洗脱液；洗脱液喷雾干燥，得到黑果枸杞提取物。
其中黑果枸杞提取物的制备方法中，所述的提取料液比1:8～1:12，低温高速剪切HSDE条件如下：15000～20000rpm，时间30-90min；离心条件如下：10000～16000r/min，时间10～30min。
所述的大孔树脂层析柱选用D101、LX-11或LX-60。
更进一步的，所述的组合物中的分数比如下：枸杞提取物1份～9份，黑果枸杞提取物1份～9份。
本发明的组合物可用于制备用于明目抗蓝光的制品。
本发明所提供的组合物具有很好的保护视力的作用，可以作为明目抗蓝光产品开发的原料。本发明通过将枸杞提取物和黑果枸杞提取物以最佳比例配比，制备了一种具有明目抗蓝光活性的枸杞和黑果枸杞的组合物，能够有效保护长期使用电子产品的人群的视力，预防眼科疾病的发生。按照现代医学解释，“上火”是过敏性应激反应，枸杞“上火”即食物过敏，主要取决于枸杞中的过敏原介质，而本发明中制备的枸杞提取物避免了食用枸杞过量导致的“上火”的问题，也解决了因服用剂量小而不能起到明目的作用的问题。
附图说明
图1为实施例1的枸杞提取物HPLC色谱图；
图2为实施例1的原花青素标准曲线图；
图3为实施例1的细胞内ROS检测图，其中A为空白对照组、B.为10μg/mL样品组、C为蓝光组，D为蓝光+样品组。
具体实施方式
本发明通过提取、分离等技术获得了枸杞提取物和黑果枸杞提取物，再将二者复配，获得了明目抗蓝光的组合物。
以下通过具体实施例对本发明进一步解释和说明。
实施例1：
1、枸杞提取物制备
将10kg枸杞粉碎，过筛至15目以下，加入亚临界萃取装置中，再加入300L的丁烷，充氮气3h后，进行亚临界萃取；萃取压力为4Mpa，萃取温度为30℃，萃取3h后脱溶剂，收集枸杞脂溶性成分，辐照灭菌，得到黄色油状的枸杞提取物。
将枸杞提取物用二氯甲烷溶解，终浓度为0.1mg/mL，吸取枸杞提取物溶液20μL注入液相色谱仪，色谱条件如下：色谱柱为WatersYMC Carotenoid(5μm,4.6×150mm)，流速0.5mL/min，波长446nm，流动相为甲醇(A):甲基叔丁基醚(B)，流动相比例：0～10min A(90～25％)：B(10～75％)，10～30min A:B＝25:75。从HPLC谱图(图1)可以看出，枸杞提取物中主要含有β-胡萝卜素、叶黄素、玉米黄质。
2、黑果枸杞提取物制备
称黑果枸杞1kg，用8L的水浸泡3小时，利用高速剪切机处理60min(19000rpm)，离心，取上清液，提取2次，得到黑果枸杞提取液。将上述黑果枸杞提取液先过10000Da的膜，去除＞10000分子量的物质，再过160Da的膜，浓缩至50mL，待用。称取1kg D101大孔吸附树脂，在水溶液中均匀混合，湿法装入一根内径15cm、柱高200cm的树脂柱中，将上述浓缩液直接泵入树脂柱，进行反复多次吸附，流速为5～10BV/h(柱体积，简称BV)，收集吸附残液；吸附结束后，用30BV的洗脱液(60％乙醇)进行洗脱，流速为5BV/h，收集洗脱液，将洗脱液喷雾干燥，得到黑果枸杞提取物652g。
以原花青素浓度(mg/mL)为横坐标，吸光度值为纵坐标绘制标准曲线(图2)，得回归方程：y＝4.4614x-0.00 45，R2＝0.9987。原花青素浓度在0～0.200mg/mL范围内与吸光度线性关系良好。精密称取适量原花青素样品，用甲醇定容于50mL容量瓶中，得样品溶液。将正丁醇与盐酸按体积比95:5混合后，取出6mL置于具塞锥形瓶中，加入0.2mL硫酸铁铵溶液和1mL样品溶液，混匀，置沸水浴回流，加热40min后，立即置冰水中冷却，在加热完毕15min后，于546nm波长处测定吸光度，计算原花青素含量＝C×T×V/M×100％(式中：C，测得原花青素含量，mg/mL；T，稀释倍数；V，定容体积，mL；M，样品质量，mg)，结果显示，本实施例中高速剪切提取的原花青素含量(3.54％)高于超声提取的原花青素含量(1.28％)和冷浸提取的原花青素含量(0.47％)
实施例2：枸杞和黑果枸杞的组合物明目抗蓝光活性测定
本发明中的明目抗蓝光的枸杞和黑果枸杞的组合物，其中枸杞提取物取2份，黑果枸杞提取物取1份；该组合物具有很好的保护视力的作用，可以作为明目抗蓝光产品开发的原料。
以枸杞和黑果枸杞的组合物为研究对象，培养ARPE-19人视网膜上皮细胞，设置以下4个组别，包含有：A.空白对照组：在同等实验条件下避光培养；B.样品组：10μg/mL枸杞和黑果枸杞的组合物无菌处理后加入到细胞中避光培养；C.蓝光组：采用波长405nm，照射强度160μW/cm 2的LED蓝光照射，连续3天照射40min后继续培养，最后一次照射完成24小时后进行检测；D.蓝光+样品组：采用10μg/mL枸杞和黑果枸杞的组合物孵育2h后进行蓝光照射，具体操作同C组。然后采用活性氧检测试剂盒检测细胞内ROS生成水平，最后在倒置荧光显微镜下观察。
结果如图3所示，空白对照组、样品组、蓝光+样品组荧光染色较弱，蓝光组荧光染色较强。与对照组相比，蓝光组细胞荧光增强，表明蓝光照射后细胞内ROS生成增加。蓝光+样品组比蓝光组的荧光染色减弱。表明浓度为10μg/mL的枸杞和黑果枸杞的组合物溶液能降低蓝光照射诱导的细胞内ROS水平。有研究发现报道LED蓝光照射能导致视网膜上皮细胞的线粒体DNA发生损伤，使细胞增殖活力降低，其机制可能与蓝光照射视网膜上皮细胞后诱导细胞内ROS及炎症因子的产生增加所致。因此，枸杞和黑果枸杞组合物具有抗蓝光明目的功效，可以作为明目抗蓝光产品开发的原料。
Claims (9)
1.一种枸杞和黑果枸杞的组合物，其特征在于，所述的组合物包含有枸杞提取物和黑果枸杞提取物；
其中枸杞提取物的制备方法如下：
将枸杞粉碎，在枸杞粉末中加入萃取溶剂后置于亚临界萃取设备中进行萃取，然后收集脂溶性成分，灭菌，得到枸杞提取物；
所述黑果枸杞提取物的制备方法，是将黑果枸杞用水为提取溶剂进行高速剪切提取，离心后，获得黑果枸杞提取液；然后将黑果枸杞提取液先用10000～100000Da的膜去除大分子物质，再用孔径大小为160～360Da的膜进行浓缩；浓缩液采用大孔树脂层析柱吸附，用乙醇溶液洗脱，得到黑果枸杞洗脱液；洗脱液喷雾干燥，得到黑果枸杞提取物。
2.如权利要求1所述的枸杞和黑果枸杞的组合物，其特征在于，所述的枸杞提取物的制备方法中，萃取溶剂为食品添加剂丙烷或丁烷。
3.如权利要求1所述的枸杞和黑果枸杞的组合物，其特征在于，所述的枸杞提取物的制备方法中，枸杞粉末与萃取溶剂质量体积比为1:10～1:40。
4.如权利要求1所述的枸杞和黑果枸杞的组合物，其特征在于，所述的枸杞提取物的制备方法中，萃取压力为1～10Mpa，萃取时间为2～5h，萃取温度为20℃～50℃。
5.如权利要求1所述的枸杞和黑果枸杞的组合物，其特征在于，所述的黑果枸杞提取物的制备方法中，所述的提取料液比1:8～1:12，低温高速剪切HSDE条件如下：15000～20000rpm，时间15-30min；离心条件如下：10000～16000r/min，时间30-90min。
6.如权利要求1所述的枸杞和黑果枸杞的组合物，其特征在于，所述的黑果枸杞提取物的制备方法中，大孔树脂层析柱选用D101、LX-11或LX-60。
7.一种枸杞和黑果枸杞的组合物，其特征在于，所述的组合物中的枸杞提取物为1份～9份，黑果枸杞提取物为1份～9份。
8.权利要求1所述的枸杞和黑果枸杞的组合物在制备用于明目抗蓝光的制品中的应用。
9.一种用于明目抗蓝光的制品，其特征在于，所述的制品中包含有药理有效浓度的权利要求1所述的枸杞和黑果枸杞的组合物。
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