CN108157970A - 蚕蛹复合油脂微胶囊的制备方法 - Google Patents
蚕蛹复合油脂微胶囊的制备方法 Download PDFInfo
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Abstract
本发明涉及一种蚕蛹复合油脂微胶囊的制备方法，采用超临界CO2萃取和梯度冷冻结晶制备高质量蚕蛹复合油脂，再通过微囊化技术制备蚕蛹复合油脂微胶囊。有益效果为：生产工艺简单，自动化高，可快速从蚕蛹油中提取α－亚麻酸含量极高的复合油脂。提取得到的复合油脂清香自然、有机溶剂残留少、α－亚麻酸含量极高、性质稳定，可有效预防及治疗高血脂症和促进肝细胞再生，预防和治疗非酒精性脂肪肝。
Description
技术领域
本发明涉及食品加工领域，具体是一种蚕蛹复合油脂微胶囊的制备方法。
背景技术
在《中药大辞典》中有如下记载：蚕蛹油可治小儿疳热、消瘦、消渴；蚕蛹油富含多种氨基酸、微量元素及n-3和n-6系列多不饱和脂肪酸，尤其含有人体所必需的多不饱和脂肪酸。目前，对蚕蛹油的开发研究，更多的是侧重降低血浆，胆固醇，改善肝脏功能的作用，增加体内二十碳五烯酸(EPA)、二十二碳六烯酸(DHA)的生成，并且具有调整血脂、抗氧化作用；同时，蚕蛹当中含有大量的氨基酸，即关注点放在了降脂方面和营养保健当中。
目前蚕蛹油提取的方法主要有溶剂浸出法和溶剂压榨法，然而这些方法都存在有一定的缺陷。蚕蛹是高蛋白、高油脂物质，粗纤维含量低，质地比较软，所以溶剂压榨提取油脂困难较大，同时蚕蛹油得率也很低。溶剂浸出法提取油脂，虽然不受油料质地软硬的影响，油料经适当的预处理后，选用合适的溶剂.，即可浸出油脂，但现在的溶剂浸出法普遍采用二氯甲烷、丙酮等毒性较大的单一溶剂提取蚕蛹油.，这样不仅污染大.，而且产率低，不利于工业化的生产；即使经过改进后的方法，(例鄂旭等利用环己烷-石油醚混合溶剂提取蚕蛹毛油)但对溶剂的需求量大，消耗时间长。
在现有技术中，不饱和脂肪酸在原料中含量一般不是很高，例如DHA，EPA在鱼油中的含量在15％-30％。不饱和脂肪酸在分离以及纯化过程中，容易受氧、热等作用而发生氧化、聚合、降解、异构化等反应，产生一些对人体有害的物质。同时，作为医药工业和保健食品原料时，必须除去原料中含有的色素和产生臭味的化合物如醛、酮类，产品内也不能残留分离过程中引入的有毒溶剂。
发明内容
本发明的目的在于提供生产工艺简单，可快速从蚕蛹油中提取α－亚麻酸含量极高的蚕蛹复合油脂微胶囊的制备方法。制备得到的复合油脂微胶囊中α－亚麻酸含量极高且性质稳定，可有效预防及治疗高血脂症和促进肝细胞再生，预防和治疗非酒精性脂肪肝。
为达到上述目的，本发明采取的技术方案为：蚕蛹复合油脂微胶囊的制备方法，采用超临界CO2萃取和梯度冷冻结晶制备高质量蚕蛹复合油脂，再通过微囊化技术制备蚕蛹复合油脂微胶囊；其中，超临界CO2萃取具体为将蚕蛹油置于超临界CO2装置中，进行超临界萃取，采用的夹带剂为含有5~10%乙酸乙酯和0.01~0.02%氧氟沙星的乙醇溶液，收集萃余液，得到初步精制油。超临界CO2萃取法是一种新型的无毒溶剂萃取技术，利用超临界CO2流体在不同温度和压力下利用对不同物质的溶解度差别将其分离，工艺简单，生产成本低，特别适用于脂溶性、高沸点、热敏性物质的提取。采用超临界CO2萃取法从蚕蛹中提取得到的复合油脂，无腥味臭味、色泽透明亮黄、无有机溶剂残留，α－亚麻酸含量高，对血脂有较好的调节作用，降低大鼠血清中TC及MDA含量、升高HDLC/TC比值、降低肝组织中MDA含量，并增加人体体内EPA和DHA合成，具有调整血脂、抗脂质过氧化作用，可有效预防及治疗高血脂症。具有促进肝细胞再生作用，降低血清谷丙转氨酶作用，通过调整脂质代谢，具有十分显著的预防和治疗非酒精性脂肪肝。
超临界萃取条件为：萃取温度为35~40℃，萃取压力为30~40MPa，萃取时间0.6~1.0h；分离釜Ⅰ压力10~12MPa，温度40~45℃；分离釜Ⅱ压力5~7MPa，温度15~20℃。上述萃取条件下游离脂肪酸、醛和酮类等杂质在超临界CO2流体中的溶解度高，油酸、亚油酸等复合油脂，特别是α－亚麻酸在超临界CO2流体中的溶解度极低，当超临界CO2流体进入解压釜中，调节到上述解压条件时杂质析出，CO2再经过冷凝器冷凝后进入CO2储罐中循环使用。
采用的夹带剂为含有5~10%乙酸乙酯和0.01~0.02%氧氟沙星的乙醇溶液。其中，氧氟沙星的组成为80~90%左旋氧氟沙星和10~20%右旋氧氟沙星。上述夹带剂中各成分相互作用，通过化学谛合作用，大大提高游离脂肪酸、醛和酮类等杂质在超临界CO2流体中的溶解度对的温度、压力的敏感程度，提高最终得到的复合油脂的纯度。在夹带剂乙醇溶液中加入0.01~0.02%氧氟沙星，特别是氧氟沙星的组成为80~90%左旋氧氟沙星和10~20%右旋氧氟沙星，能把提高夹带剂的抑菌性，减缓提取过程中复合油脂的腐败速度。上述特定配比的氧氟沙星可缠绕在复合油脂的双键处，通过范德华力和氢键作用，将其与超临界CO2流体分子隔离开，降低提取的复合油脂对氧的敏感程度，提高超临界CO2的萃取效率。
梯度冷冻结晶：初步精制油与质量分数为20~30％的尿素乙醇溶液按质量比为1：7~12混合均匀，梯度冷冻结晶；抽滤，滤液重复梯度冷冻一次，过滤，滤液用HCl溶液酸化至pH值为2～3，用石油醚萃取，分离石油醚相，用蒸馏水冲洗石油醚相至中性，干燥石油醚相，减压蒸馏回收石油醚，制备得到蚕蛹油复合油脂。梯度冷冻干燥：8~10℃，2.5~3h；4~5℃，1.5~2h；1~1.5℃，0.8~1h；-2~-1.5℃，0.5~0.8h；-4~-3.5℃，0.5~0.8h；-6~-5.5℃，0.5~0.8h；-8~-7.5℃，0.5~0.8h；-12~-11.5℃，0.5~0.8h。利用低温下不同的脂肪酸或脂肪酸盐在有机溶剂中溶解度不同来进行分离纯化。通过降温就可过滤除去其中大量的饱和脂肪酸和部分单复合油脂，从而获得所需的多价复合油脂。
微囊化技术为：在蚕蛹油复合油脂中加入油脂0.5~0.8倍体积的MCT和7~10倍体积的Cremophor EL 35，超声混合后，在磁力搅拌下滴加混合液0.1~0.4倍的无水乙醇，得到蚕蛹复合油脂微胶囊。超声波频率为15~30kHZ，功率为150~200W，超声时间为5~10min。无水乙醇的滴加速度为1~2滴/秒。上述操作制备的蚕蛹复合油脂微胶囊为微乳液，MCT和Cremophor EL 35起到稳定乳化作用，形成微乳液为复乳结构（W1/O/W2）结构，这种三相结构性质的存在使得复乳结构中的活性成分性质稳定，提高蚕蛹复合油脂，特别是α－亚麻酸的稳定性，延长其保质期。
与现有技术相比，本发明的优点在于：生产工艺简单，自动化高，可快速从蚕蛹油中提取α－亚麻酸含量极高的复合油脂。本发明超临界CO2萃取采用的夹带剂不仅能大大提高游离脂肪酸、醛和酮类等杂质在超临界CO2流体中的溶解度对的温度、压力的敏感程度，提高最终得到的复合油脂的纯度，还能降低提取的复合油脂对氧的敏感程度，提高超临界CO2的萃取效率。提取得到的复合油脂清香自然、有机溶剂残留少、α－亚麻酸含量极高、性质稳定，可有效预防及治疗高血脂症和促进肝细胞再生，预防和治疗非酒精性脂肪肝。
具体实施方式
下面通过实施例对本发明方案作进一步说明：
实施例1：
蚕蛹复合油脂微胶囊的制备方法，最优选步骤为：
1）超临界萃取：将蚕蛹油置于超临界CO2装置中，进行超临界萃取，采用的夹带剂为含有7%乙酸乙酯和0.01%氧氟沙星的乙醇溶液，收集萃余液，得到精制油。其中，氧氟沙星的组成为90%左旋氧氟沙星和10%右旋氧氟沙星。超临界萃取条件为：萃取温度为37℃，萃取压力为35MPa，萃取时间0.8h；分离釜Ⅰ压力11MPa，温度42℃；分离釜Ⅱ压力6MPa，温度16℃。上述萃取条件下游离脂肪酸、醛和酮类等杂质在超临界CO2流体中的溶解度高，油酸、亚油酸等复合油脂，特别是α－亚麻酸在超临界CO2流体中的溶解度极低，当超临界CO2流体进入解压釜中，调节到上述解压条件时杂质析出，CO2再经过冷凝器冷凝后进入CO2储罐中循环使用。提取得到的复合油脂，无腥味臭味、色泽透明亮黄、无有机溶剂残留，α－亚麻酸含量高，对血脂有较好的调节作用，降低大鼠血清中TC及MDA含量、升高HDLC/TC比值、降低肝组织中MDA含量，并增加人体体内EPA和DHA合成，具有调整血脂、抗脂质过氧化作用，可有效预防及治疗高血脂症。具有促进肝细胞再生作用，降低血清谷丙转氨酶作用，通过调整脂质代谢，具有十分显著的预防和治疗非酒精性脂肪肝；
2）梯度冷冻结晶:初步精制油与质量分数为25％的尿素乙醇溶液按质量比为1：10混合均匀，梯度冷冻干燥：9℃，3h；4.5℃，1.5h；1℃，1h；-2℃，0.5h；-4℃，0.5h；-6℃，0.8h；-8℃，0.8h；-12℃，0.5h。抽滤，滤液重复梯度冷冻一次，过滤，滤液用质量分数为10％的HCl溶液酸化至pH值为2～3，用石油醚萃取，分离石油醚相，用蒸馏水冲洗石油醚相至中性，按石油醚相与无水硫酸钠的质量比为25∶1将石油醚相通过装有无水硫酸钠的漏斗过滤干燥，再用减压蒸发器40～50℃、-0.08MPa减压蒸馏，回收石油醚，制备成蚕蛹油复合油脂；
3）微囊化：在蚕蛹油复合油脂中加入油脂0.6倍体积的MCT和8倍体积的Cremophor EL35，超声混合后，在磁力搅拌下滴加混合液0.3倍的无水乙醇，得到蚕蛹复合油脂微胶囊。超声波频率为20kHZ，功率为200W，超声时间为8min。无水乙醇的滴加速度为1~2滴/秒。
实施例2：
蚕蛹复合油脂微胶囊的制备方法，具体操作为：
1）超临界萃取：将蚕蛹油置于超临界CO2装置中，进行超临界萃取，采用的夹带剂为含有5%乙酸乙酯和0.02%氧氟沙星的乙醇溶液，收集萃余液，得到精制油。其中，氧氟沙星的组成为80%左旋氧氟沙星和20%右旋氧氟沙星。超临界萃取条件为：萃取温度为35℃，萃取压力为40MPa，萃取时间1.0h；分离釜Ⅰ压力10MPa，温度40℃；分离釜Ⅱ压力5MPa，温度15℃。上述萃取条件下游离脂肪酸、醛和酮类等杂质在超临界CO2流体中的溶解度高，油酸、亚油酸等复合油脂，特别是α－亚麻酸在超临界CO2流体中的溶解度极低，当超临界CO2流体进入解压釜中，调节到上述解压条件时杂质析出，CO2再经过冷凝器冷凝后进入CO2储罐中循环使用。提取得到的复合油脂，无腥味臭味、色泽透明亮黄、无有机溶剂残留，α－亚麻酸含量高，对血脂有较好的调节作用，降低大鼠血清中TC及MDA含量、升高HDLC/TC比值、降低肝组织中MDA含量，并增加人体体内EPA和DHA合成，具有调整血脂、抗脂质过氧化作用，可有效预防及治疗高血脂症。具有促进肝细胞再生作用，降低血清谷丙转氨酶作用，通过调整脂质代谢，具有十分显著的预防和治疗非酒精性脂肪肝；
2）梯度冷冻结晶:初步精制油与质量分数为25％的尿素乙醇溶液按质量比为1：10混合均匀，梯度冷冻干燥：9℃，3h；4.5℃，1.5h；1℃，1h；-2℃，0.5h；-4℃，0.5h；-6℃，0.8h；-8℃，0.8h；-12℃，0.5h。抽滤，滤液重复梯度冷冻一次，过滤，滤液用质量分数为10％的HCl溶液酸化至pH值为2～3，用石油醚萃取，分离石油醚相，用蒸馏水冲洗石油醚相至中性，按石油醚相与无水硫酸钠的质量比为25∶1将石油醚相通过装有无水硫酸钠的漏斗过滤干燥，再用减压蒸发器40～50℃、-0.08MPa减压蒸馏，回收石油醚，制备成蚕蛹油复合油脂；
3）微囊化：在蚕蛹油复合油脂中加入油脂0.6倍体积的MCT和8倍体积的Cremophor EL35，超声混合后，在磁力搅拌下滴加混合液0.3倍的无水乙醇，得到蚕蛹复合油脂微胶囊。超声波频率为20kHZ，功率为200W，超声时间为8min。无水乙醇的滴加速度为1~2滴/秒。
实施例3：
蚕蛹复合油脂微胶囊的制备方法，具体操作为：
1）超临界萃取：将蚕蛹油置于超临界CO2装置中，进行超临界萃取，采用的夹带剂为含有6%乙酸乙酯和0.01%氧氟沙星的乙醇溶液，收集萃余液，得到精制油。其中，氧氟沙星的组成为80%左旋氧氟沙星和20%右旋氧氟沙星。超临界萃取条件为：萃取温度为37℃，萃取压力为40MPa，萃取时间1.0h；分离釜Ⅰ压力10MPa，温度40℃；分离釜Ⅱ压力5MPa，温度17℃。上述萃取条件下游离脂肪酸、醛和酮类等杂质在超临界CO2流体中的溶解度高，油酸、亚油酸等复合油脂，特别是α－亚麻酸在超临界CO2流体中的溶解度极低，当超临界CO2流体进入解压釜中，调节到上述解压条件时杂质析出，CO2再经过冷凝器冷凝后进入CO2储罐中循环使用。提取得到的复合油脂，无腥味臭味、色泽透明亮黄、无有机溶剂残留，α－亚麻酸含量高，对血脂有较好的调节作用，降低大鼠血清中TC及MDA含量、升高HDLC/TC比值、降低肝组织中MDA含量，并增加人体体内EPA和DHA合成，具有调整血脂、抗脂质过氧化作用，可有效预防及治疗高血脂症。具有促进肝细胞再生作用，降低血清谷丙转氨酶作用，通过调整脂质代谢，具有十分显著的预防和治疗非酒精性脂肪肝；
2）梯度冷冻结晶:初步精制油与质量分数为25％的尿素乙醇溶液按质量比为1：10混合均匀，梯度冷冻干燥：9℃，3h；4.5℃，1.5h；1℃，1h；-2℃，0.5h；-4℃，0.5h；-6℃，0.8h；-8℃，0.8h；-12℃，0.5h。抽滤，滤液重复梯度冷冻一次，过滤，滤液用质量分数为10％的HCl溶液酸化至pH值为2～3，用石油醚萃取，分离石油醚相，用蒸馏水冲洗石油醚相至中性，按石油醚相与无水硫酸钠的质量比为25∶1将石油醚相通过装有无水硫酸钠的漏斗过滤干燥，再用减压蒸发器40～50℃、-0.08MPa减压蒸馏，回收石油醚，制备成蚕蛹油复合油脂；
3）微囊化：在蚕蛹油复合油脂中加入油脂0.6倍体积的MCT和8倍体积的Cremophor EL35，超声混合后，在磁力搅拌下滴加混合液0.3倍的无水乙醇，得到蚕蛹复合油脂微胶囊。超声波频率为20kHZ，功率为200W，超声时间为8min。无水乙醇的滴加速度为1~2滴/秒。
实施例4：降血脂效果测试
Wister雄性小鼠100只，体重18~24g，基础饲料饲养1周，以适应环境，然后按体重随机分成2个处理，灌喂号申请公布号为CN101892119A从蚕蛹油中提取的复合油脂制品和本发明实施例1提取的成品油。对照组:[高脂饲料+申请公布号为CN101892119A从蚕蛹油中提取的复合油脂制品(1.5g/kg)]；试验组：[高脂饲料+本发明实施例1制备的蚕蛹复合油脂微胶囊(1.5g/kg)]；空白对照组：[高脂饲料+生理盐水]。各组试验小鼠均自由饮水，在自然光照、室温25℃条件下饲养，每日灌胃给药2次，0.6mL/次。高脂饲料:在基础饲料中加入1%胆固醇、0.3%胆酸、10%猪油、4%吐温80制成。
动物血样采集和检测：试验结束前1天所有小鼠禁食24h后摘眼球取血，血液样品5000r/min离心10min，取血清检测TC、TG、LDL-c、HDL-c。
统计方法：试验数据以x±s表示，采用SPSS10.0软件进行组间单因素方差分析，多重比较用LSD法和S-N-K法。
复合油脂制品对小鼠血脂的影响（x±s）
| 组别 | 总胆固醇 | 甘油三酯 | 低密度脂蛋白 | 高密度脂蛋白 |
| 空白对照组 | 3.351±0.335a | 1.827±0.416a | 2.682±0.113a | 0.946±0.0641d |
| 对照组 | 2.225±0.536b | 1.342±0.097a | 0.785±0.132b | 1.396±0.138d |
| 试验组 | 0.526±0.038b | 0.298±0.195b | 0.704±0.083a | 1.148±0.075d |
由上表可知，本发明制备的蚕蛹复合油脂微胶囊降低总胆固醇和甘油三酯的效果要远优于现有技术，具有显著的进步。
本发明的操作步骤中的常规操作为本领域技术人员所熟知，在此不进行赘述。
以上所述的实施例对本发明的技术方案进行了详细说明，应理解的是以上所述仅为本发明的具体实施例，并不用于限制本发明，凡在本发明的原则范围内所做的任何修改、补充或类似方式替代等，均应包含在本发明的保护范围之内。
Claims (8)
1.蚕蛹复合油脂微胶囊的制备方法，其特征在于采用超临界CO2萃取和梯度冷冻结晶制备高质量蚕蛹复合油脂，再通过微囊化技术制备蚕蛹复合油脂微胶囊；其中，超临界CO2萃取具体为将蚕蛹油置于超临界CO2装置中，进行超临界萃取，采用的夹带剂为含有5~10%乙酸乙酯和0.01~0.02%氧氟沙星的乙醇溶液，收集萃余液，得到初步精制油。
2.根据权利要求1所述的蚕蛹复合油脂微胶囊的制备方法，其特征在于：所述的超临界萃取条件为：萃取温度为35~40℃，萃取压力为30~40MPa，萃取时间0.6~1.0h；分离釜Ⅰ压力10~12MPa，温度40~45℃；分离釜Ⅱ压力5~7MPa，温度15~20℃。
3.根据权利要求1所述的蚕蛹复合油脂微胶囊的制备方法，其特征在于：所述的氧氟沙星的组成为80~90%左旋氧氟沙星和10~20%右旋氧氟沙星。
4.根据权利要求1所述的蚕蛹复合油脂微胶囊的制备方法，其特征在于：所述的梯度冷冻结晶：初步精制油与质量分数为20~30％的尿素乙醇溶液按质量比为1：7~12混合均匀，梯度冷冻结晶；抽滤，滤液重复梯度冷冻一次，过滤，滤液用HCl溶液酸化至pH值为2～3，用石油醚萃取，分离石油醚相，用蒸馏水冲洗石油醚相至中性，干燥石油醚相，减压蒸馏回收石油醚，制备得到蚕蛹油复合油脂。
5.根据权利要求4所述的蚕蛹复合油脂微胶囊的制备方法，其特征在于：所述的梯度冷冻干燥：8~10℃，2.5~3h；4~5℃，1.5~2h；1~1.5℃，0.8~1h；-2~-1.5℃，0.5~0.8h；-4~-3.5℃，0.5~0.8h；-6~-5.5℃，0.5~0.8h；-8~-7.5℃，0.5~0.8h；-12~-11.5℃，0.5~0.8h。
6. 根据权利要求1所述的蚕蛹复合油脂微胶囊的制备方法，其特征在于：所述的微囊化技术为：在蚕蛹油复合油脂中加入油脂0.5~0.8倍体积的MCT和7~10倍体积的CremophorEL 35，超声混合后，在磁力搅拌下滴加混合液0.1~0.4倍的无水乙醇，得到蚕蛹复合油脂微胶囊。
7.根据权利要求6所述的蚕蛹复合油脂微胶囊的制备方法，其特征在于：所述的超声波频率为15~30kHZ，功率为150~200W，超声时间为5~10min。
8.根据权利要求6所述的蚕蛹复合油脂微胶囊的制备方法，其特征在于：所述的无水乙醇的滴加速度为1~2滴/秒。
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