CN112056544A - 一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法 - Google Patents
一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法 Download PDFInfo
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Abstract
本发明提供了一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，属于农产品加工技术领域。该方法以温州蜜柑皮作为原料，采用超高压辅助酶法对果胶进行低酯化改性，制备高凝胶性和乳化稳定性的低酯果胶，可稳定负载类胡萝卜素等脂溶性营养成分；同时利用钙‑糖醇诱导果胶乳液凝胶负载脂溶性活性成分，制得的产品稳定性好、分散性高、生物利用度高，适用于低热量功能性半固态食品的开发。
Description
技术领域
本发明涉及凝胶加工技术领域，具体而言，涉及一种乳液凝胶，尤其涉及一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法。
背景技术
类胡萝卜素、黄酮等天然脂溶性活性成分，具有抗氧化、抗病毒、增强人体免疫力和防癌抗癌等多种生理功能，但由于脂溶性营养素在消化过程中溶解度低、稳定性差，易发生转化或降解，从而导致膳食脂溶性营养素的稳定性和生物利用度较低。
果胶是广泛存在于植物细胞壁中的多糖物质，具有优良的凝胶、乳化、稳定等功能特性。FAO/WHO食品添加剂联合委员会将果胶推荐为安全的天然食品添加剂，且不限制每日允许摄入量。根据酯化度的高低，可将果胶分为高酯果胶和低酯果胶两大类，天然果胶大都为高酯果胶，按“糖-酸凝胶机制”需在可溶性固形物高于55％、pH值3.0左右凝胶，主要用于高糖食品生产；天然低酯果胶很少，主要与体系中Ca2+交联并通过钙桥形成“蛋盒型”结构凝胶。由于在低糖或无糖条件下可实现凝胶，低酯果胶被广泛用于低糖果酱、果冻、酸奶等保健食品的生产。
乳液凝胶(emulsiongel)是在稳定剂存在下由分散相内相与连续相外相构成的一种被乳状液滴填充的凝胶网状结构。乳液凝胶作为一个重要的产品体系在基础研究及实际应用中受到越来越多的关注，可有效提高营养素在食物中的分散性、稳定性，同时在胃肠消化过程中具有缓释作用，利于机体对营养素活性成分的吸收利用。乳液凝胶可作为脂肪替代物，能够有效降低饱和脂肪酸的含量，提高多不饱和脂肪酸、人体必需氨基酸、矿物质等含量；还可作为功能因子的传递体系，负载类胡萝卜素、植物甾醇、鱼油、精油和脂溶性维生素等活性物质，并能显著的提高生物活性因子的稳定性。已有研究表明乳液凝胶可显著提高与其共同摄入的活性成分生物利用率。
由于果胶同时具有乳化和凝胶功能，因此果胶基乳液凝胶具有潜在的递送脂溶性营养素的功能。目前，对于低酯果胶乳液凝胶负载类胡萝卜素等脂溶性活性成分的体系制备及其在在低热量食品中的应用鲜有报道。
发明内容
鉴于现有技术的不足，本发明人利用钙-糖醇诱导果胶乳液凝胶负载脂溶性活性成分，制得的产品稳定性好、分散性高、生物利用度高，适用于低热量功能性半固态食品的开发。
为了实现上述发明目的，发明人结合多年来乳液凝胶技术的研究经验，并通过大量实验反复研究，最终获得了以下技术方案：一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，该方法包括以下步骤：
1)果胶低酯化改性：将提取的天然果胶溶解于水中配制成质量浓度0.05-2％的果胶溶液，添加果胶甲基酯酶使其浓度为50-500PEU/mL，密封包装后采用高静压处理，压力200-400MPa，温度30-40℃，时间8-12min，然后立即沸水处理3-8min，离心，取上清液，加无水乙醇混匀，离心，所得沉淀干燥得高乳化性的低酯果胶；
2)果胶乳液制备：取步骤1)得到的低酯果胶，采用柠檬酸盐缓冲液调节pH，作为水相；将脂溶性活性成分溶于大豆油中，作为油相；将油相与水相混合，超声波乳化8-13min，所述超声波的频率为18-22kHz，功率为350-450W；
3)果胶乳液凝胶制备：取步骤2)制备的乳液，加入钙盐与糖醇混合配制的乳液凝胶诱导剂，在80-90℃条件下均匀混合后，2-6℃冷藏18-30h后得到果胶乳液凝胶。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中提取所述天然果胶的原料为温州蜜柑皮。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中天然果胶的提取方法为：取温州蜜柑皮热烫钝酶后冻干磨粉，加入1M的柠檬酸盐缓冲液中，调pH＝1.3-2.0，80-90℃水浴提取50-90min，离心，加无水乙醇混匀，离心，所得沉淀为天然果胶。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中步骤1)中所述低酯果胶的酯化度为20-40％。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中步骤2)中所述水溶液中低酯果胶的浓度为0.5-2.0％，pH为2.0-4.0。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中步骤2)制备的果胶乳液中，油相与水相比例为1：9。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中步骤3)中所述的糖醇选自如下的一种或多种：赤藓糖醇、山梨糖醇、木糖醇。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中步骤3)中所述的糖醇为赤藓糖醇。
进一步优选地，如上所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其中步骤3)中所述的钙盐为氯化钙、碳酸钙、柠檬酸钙、草酸钙中的至少一种。
与现有技术相比，本发明提供的一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法具有如下优点和显著进步性：
(1)提取天然果胶的原料采用温州蜜柑皮，果胶乳化性显著优于商业柑橘果胶；
(2)采用超高压辅助酶法对果胶进行低酯化改性，制备高凝胶性和乳化稳定性的低酯果胶，可稳定负载类胡萝卜素等脂溶性营养成分；
(3)采用钙-糖醇联合诱导果胶乳液进行凝胶化，形成的产品具有更好的硬度和负载稳定性，且热量更低，可用于低热量功能性半固态食品的生产。
具体实施方式
以下采用实施例进一步描述本发明方法的实施过程和有益效果，试验例仅用于例证的目的，不限制本发明的保护范围，同时本领域普通技术人员根据实施例所做的显而易见的改变也包含在本发明范围之内。
实施例一
1)果胶提取：将温州蜜柑皮热烫钝酶，条件是100℃，5min，然后冻干磨粉，取10g温州蜜柑皮粉与1M的柠檬酸-柠檬酸钠缓冲液按1：40混合，然后调整pH为1.4，采用85℃水浴提取70min，离心后2倍体积无水乙醇沉淀、离心、干燥备用；
2)果胶低酯化改性：将步骤1)提取的果胶配制1％(W/W)果胶溶液，添加果胶甲基酯酶使其浓度为100PEU/mL，密封包装后采用高静压处理(400MPa/30℃/8min)后立即沸水处理5min，离心后2倍体积无水乙醇沉淀、离心、干燥,得低酯柑橘果胶；
3)果胶乳液制备：将步骤2)制备的低酯柑橘果胶配制浓度为1.0％(W/W)的水溶液，采用柠檬酸-柠檬酸钠缓冲液调节pH为3.0，作为水相；取叶黄素溶于大豆油中，以油相：水相按1：9的体积比混合，超声波乳化(20kHz/400W/10min)；
4)果胶乳液凝胶：将步骤3)制备的乳液与氯化钙(40mg/g)与赤藓糖醇水溶液(10％,w/w)，在90℃温度条件下均匀混合后，4℃冷藏24h后得到稳定的产品，硬度42.5g、叶黄素负载率98.3％；体外释放试验显示，模拟胃液体系中叶黄素的释放率仅为5.1％，而模拟肠液体系中2h后释放率75％。
实施例二
1)果胶提取：将温州蜜柑皮热烫钝酶，条件是100℃，5min，然后冻干磨粉，取10g温州蜜柑皮粉与1M的柠檬酸-柠檬酸钠缓冲液按1：40混合，然后调整pH为1.4，采用85℃水浴提取70min，离心后2倍体积无水乙醇沉淀、离心、干燥备用；
2)果胶低酯化改性：将步骤1)提取的果胶配制1％(W/W)果胶溶液，添加果胶甲基酯酶使其浓度为100PEU/mL，密封包装后采用高静压处理(200MPa/40℃/12min)后立即沸水处理5min，离心后2倍体积无水乙醇沉淀、离心、干燥,得低酯柑橘果胶；
3)果胶乳液制备：将步骤2)制备的低酯柑橘果胶配制浓度为1.0％(W/W)的水溶液，采用柠檬酸-柠檬酸钠缓冲液调节pH为3.0，作为水相；取姜黄素溶于大豆油中，以油相：水相按1：9的体积比混合，超声波乳化(20kHz/400W/10min)；
4)果胶乳液凝胶：将步骤3)制备的乳液与氯化钙(30mg/g)与赤藓糖醇水溶液(15％,w/w)，在90℃温度条件下均匀混合后，4℃冷藏24h后得到稳定的产品，硬度40.5g、姜黄素负载率98.7％；体外释放试验显示，模拟胃液体系中叶黄素的释放率仅为4.6％，而模拟肠液体系中2h后释放率72％。
实施例三
1)果胶提取：将温州蜜柑皮热烫钝酶，条件是100℃，5min，然后冻干磨粉，取10g温州蜜柑皮粉与1M的柠檬酸-柠檬酸钠缓冲液按1：40混合，然后调整pH为1.4，采用85℃水浴提取70min，离心后2倍体积无水乙醇沉淀、离心、干燥备用；
2)果胶低酯化改性：将步骤1)提取的果胶配制1％(W/W)果胶溶液，添加果胶甲基酯酶使其浓度为100PEU/mL，密封包装后采用高静压处理(300MPa/30℃/10min)后立即沸水处理5min，离心后2倍体积无水乙醇沉淀、离心、干燥,得低酯柑橘果胶；
3)果胶乳液制备：将步骤2)制备的低酯柑橘果胶配制浓度为1.0％(W/W)的水溶液，采用柠檬酸-柠檬酸钠缓冲液调节pH为3.0，作为水相；取β-胡萝卜素溶于大豆油中，以油相：水相按1：9的体积比混合，超声波乳化(20kHz/400W/10min)；
4)果胶乳液凝胶：将步骤3)制备的乳液与氯化钙(50mg/g)与赤藓糖醇水溶液(10％,w/w)，在90℃温度条件下均匀混合后，4℃冷藏24h后得到稳定的产品，硬度39.2g、β-胡萝卜素负载率98.0％；体外释放试验显示，模拟胃液体系中叶黄素的释放率仅为6.3％，而模拟肠液体系中2h后释放率80％。
对比例一：温州蜜柑果胶替换为市售柑橘果胶
1)果胶采用购买的市售柑橘果胶(sigmap5400)；
2)果胶低酯化改性：将步骤1)的果胶配制1％(W/W)果胶溶液，添加果胶甲基酯酶使其浓度为100PEU/mL，密封包装后采用高静压处理(300MPa/30℃/10min)后立即沸水处理5min，离心后2倍体积无水乙醇沉淀、离心、干燥,得低酯柑橘果胶；
3)果胶乳液制备：将步骤2)制备的低酯柑橘果胶配制浓度为1.0％(W/W)的水溶液，采用柠檬酸-柠檬酸钠缓冲液调节pH为3.0，作为水相；取叶黄素溶于大豆油中，以油相：水相按1：9的体积比混合，超声波乳化(20kHz/400W/10min)；
4)果胶乳液凝胶：将步骤3)制备的乳液与氯化钙(50mg/g)与赤藓糖醇水溶液(10％,w/w)，在90℃温度条件下均匀混合后，4℃冷藏24h后得到稳定的产品，硬度29.4g、叶黄素负载率75.0％；体外释放试验显示，模拟胃液体系中叶黄素的释放率为40.1％。
对比例二：果胶低酯化方法替换为常压酶法
1)果胶提取：将温州蜜柑皮热烫钝酶，条件是100℃，5min，然后冻干磨粉，取10g温州蜜柑皮粉与1M的柠檬酸-柠檬酸钠缓冲液按1：40混合，然后调整pH为1.4，采用85℃水浴提取70min，离心后2倍体积无水乙醇沉淀、离心、干燥备用；
2)果胶低酯化改性：将步骤1)提取的果胶配制1％(W/W)果胶溶液，添加果胶甲基酯酶使其浓度为100PEU/mL，常压30℃低酯反应2h后立即沸水处理5min，离心后2倍体积无水乙醇沉淀、离心、干燥，得低酯柑橘果胶；
3)果胶乳液制备：将步骤2)制备的低酯柑橘果胶配制浓度为1.0％(W/W)的水溶液，采用柠檬酸-柠檬酸钠缓冲液调节pH为3.0，作为水相；取叶黄素溶于大豆油中，以油相：水相按1：9的体积比混合，超声波乳化(20kHz/400W/10min)；
4)果胶乳液凝胶：将步骤3)制备的乳液与氯化钙(50mg/g)与赤藓糖醇水溶液(10％,w/w)，在90℃温度条件下均匀混合后，4℃冷藏24h后得到稳定的产品，硬度32.7g、叶黄素负载率88.2％；体外释放试验显示，模拟胃液体系中叶黄素的释放率为36.2％。
对比例三：乳液凝胶诱导剂替换为单独使用氯化钙溶液
1)果胶提取：将温州蜜柑皮热烫钝酶，条件是100℃，5min，然后冻干磨粉，取10g温州蜜柑皮粉与1M的柠檬酸-柠檬酸钠缓冲液按1：40混合，然后调整pH为1.4，采用85℃水浴提取70min，离心后2倍体积无水乙醇沉淀、离心、干燥备用；
2)果胶低酯化改性：将步骤1)提取的果胶配制1％(W/W)果胶溶液，添加果胶甲基酯酶使其浓度为100PEU/mL，常压30℃低酯反应2h后立即沸水处理5min，离心后2倍体积无水乙醇沉淀、离心、干燥，得低酯柑橘果胶；
3)果胶乳液制备：将步骤2)制备的低酯柑橘果胶配制浓度为1.0％(W/W)的水溶液，采用柠檬酸-柠檬酸钠缓冲液调节pH为3.0，作为水相；取叶黄素溶于大豆油中，以油相：水相按1：9的体积比混合，超声波乳化(20kHz/400W/10min)；
4)果胶乳液凝胶：将步骤3)制备的乳液与氯化钙(50mg/g)水溶液，在90℃温度条件下均匀混合后，4℃冷藏24h后得到稳定的产品，硬度31.8g、叶黄素负载率90.5％；体外释放试验显示，模拟胃液体系中叶黄素的释放率为37.5％。
Claims (8)
1.一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其特征在于，该方法包括以下步骤：
1)果胶低酯化改性：将提取的天然果胶溶解于水中配制成质量浓度0.05-2％的果胶溶液，添加果胶甲基酯酶使其浓度为50-500PEU/mL，密封包装后采用高静压处理，压力200-400MPa，温度30-40℃，时间8-12min，然后立即沸水处理3-8min，离心，取上清液，加无水乙醇混匀，离心，所得沉淀干燥得高乳化性的低酯果胶；
2)果胶乳液制备：取步骤1)得到的低酯果胶，采用柠檬酸盐缓冲液调节pH，作为水相；将脂溶性活性成分溶于大豆油中，作为油相；将油相与水相混合，超声波乳化8-13min，所述超声波的频率为18-22kHz，功率为350-450W；
3)果胶乳液凝胶制备：取步骤2)制备的乳液，加入钙盐与糖醇混合配制的乳液凝胶诱导剂，在80-90℃条件下均匀混合后，2-6℃冷藏18-30h后得到果胶乳液凝胶。
2.根据权利要求1所述负载类胡萝卜素的果胶乳液凝胶的制备方法，其特征在于，提取所述天然果胶的原料为温州蜜柑皮。
3.根据权利要求2所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其特征在于，所述的天然果胶的提取方法为：取温州蜜柑皮热烫钝酶后冻干磨粉，加入1M的柠檬酸盐缓冲液中，调pH＝1.3-2.0，80-90℃水浴提取50-90min，离心，加无水乙醇混匀，离心，所得沉淀为天然果胶。
4.根据权利要求1所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其特征在于，步骤1)中所述水溶液中低酯果胶的酯化度为20％-40％。
5.根据权利要求1所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其特征在于，步骤2)中所述水溶液中低酯果胶的浓度为0.5-2.0％，pH为2.0-4.0。
6.根据权利要求1所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其特征在于，步骤2)制备的果胶乳液中，油相与水相比例为1：9。
7.根据权利要求1所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其特征在于，步骤3)中所述的糖醇选自如下的一种或多种：赤藓糖醇、山梨糖醇、木糖醇，浓度为10-15％；所述的钙盐为氯化钙、碳酸钙、柠檬酸钙、草酸钙中的至少一种，浓度为20-50mg/g。
8.根据权利要求1所述一种可稳定负载脂溶性活性成分的果胶乳液凝胶的制备方法，其特征在于，步骤3)中所述的糖醇为赤藓糖醇。
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