CN110498935B - 一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液及其制备方法 - Google Patents
一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液及其制备方法 Download PDFInfo
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Abstract
本发明属于乳液技术领域,公开了一种大豆分离蛋白‑果胶复合物稳定槲皮素的高内相乳液及其制备方法,所述方法包括将大豆分离蛋白溶于蒸馏水中,获得大豆分离蛋白溶液;将果胶溶于蒸馏水中,调节pH,获得果胶溶液;将大豆分离蛋白溶液与果胶溶液混合;调节溶液的pH为3.0~11.0;所得溶液与含槲皮素的植物油按比例混合后进行超声处理,离心后获得大豆分离蛋白‑果胶复合物稳定槲皮素的高内相乳液。本发明无需任何表面活性剂,所得水包油型乳液粒径小、粘弹性好、稳定性高,对槲皮素具有优异的保护或控释效果,并且本发明的制备方法对仪器要求低,方法简单且易于操作,在新型营养物质输送载体及食品结构修饰基料应用方面具有较好的前景。
Description
技术领域
本发明涉及乳液制备技术领域,更具体的说是涉及一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液及其制备方法。
背景技术
槲皮素是自然界广泛存在的多醇羟基黄酮类化合物,其酚羟基活络位点赋予了槲皮素抗癌、抗炎、抗氧化性等生理功能,然而槲皮素水溶性极差,遇光不稳定,具有较差的理化及药代动力学性质,静脉给药在体内难以运输以及口服生物利用度低,大大限制了槲皮素在食品工业、医药工业上的应用。因此,如何有效包埋槲皮素来提高其稳定性并以此提高槲皮素的生物利用度成为问题,面对此类问题,传统的方法是改变其化学结构或添加表面活性剂,但此种方法难以保障产物安全性,而构建食物蛋白-多糖稳定的乳液输送体系是提高疏水性活性成分的生物利用度及稳定性的一个重要途径,并且食物蛋白-多糖乳液本身是一类重要的食品基质地,具有良好的营养价值。
中国发明专利申请201610375510.6公开了一种可食性蛋白稳定Pickering乳液的制备方法,该申请是通过添加小麦醇溶蛋白-壳聚糖来稳定乳液,不含任何表面活性剂,但是该方法制备出的乳液粒径大,所选用的基材是小麦醇溶蛋白,小麦醇溶蛋白不耐强酸,荷载的活性物质到达胃部就已经释放,不能在肠释放,只适合胃部靶向缓释或者用于外部涂抹。
中国发明专利申请201611004761.X公开了利用玉米醇溶蛋白制备高内相凝胶状型乳液的方法,但这种方法建立在首先通过繁琐的化学或物理方法处理(如反溶剂法)制备蛋白胶体颗粒的基础上,操作不便,成本较高,且玉米醇溶蛋白等电点在6.2左右,具有较强的疏水性,导致在形成高内相乳液的过程中极易发生相反转,因此,利用天然或简单处理的蛋白-多糖制备新型高内相水包油型乳液具有更重要的意义。
大豆分离蛋白是世界上产量最高的植物蛋白质之一,具有优良的功能特性,包括溶解性、乳化性、起泡性、凝胶性和聚集性等,具有良好的生物相容性,被广泛应用于食品加工中,改善食品的质构特性;果胶是一种天然的植物多糖,存在于所有的高等植物中,具有良好的乳化、增稠、稳定和凝胶作用,能够降低血糖和胆固醇,有较好的生物降解性,被广泛应用于食品添加剂、药品和化妆品行业。
因此,如何构建大豆分离蛋白-果胶复合物稳定高内相乳液对活性物质槲皮素包埋输送,在食品药品领域的发展具有重要的意义。
发明内容
有鉴于此,本发明提供了一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法,制备得到的乳液将液态油转变成固态油,具有粘弹性,无表面活性剂的添加且能长时间稳定而不破乳反乳,大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液贮存稳定性强,实现活性物质的有效释放,可应用于食品和医药领域。
为了达到上述目的,本发明采用如下技术方案:
一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法,包括以下步骤:
(1)将大豆分离蛋白溶于蒸馏水中,获得大豆分离蛋白溶液;
(2)将果胶溶于蒸馏水中,调节pH为2.0~4.0,获得果胶溶液;
(3)将所述大豆分离蛋白溶液与所述果胶溶液混合,调节混合溶液的pH为3.0~11.0,得到溶液体系;
(4)将所述溶液体系与含槲皮素的植物油混合,进行超声均质乳化,离心后获得大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(1)中所述大豆分离蛋白溶液的质量浓度为0.2%~5.0%;
进一步优选的,步骤(1)中所述大豆分离蛋白溶液的质量浓度为5.0%。
上述技术方案的有益效果是:若大豆分离蛋白浓度太低,颗粒不足以稳定油滴界面,油滴易于发生融合,导致粒径增大,上述大豆分离蛋白溶液的浓度可以保持粒径在合适范围内。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(2)中所述果胶溶液的质量浓度为0.1%~2%;
进一步优选的,步骤(2)中所述果胶溶液的质量浓度为1%。
上述技术方案的有益效果是:在低果胶浓度下,吸附到蛋白表面的果胶颗粒较少,因此复合颗粒的疏水性较强,在乳化过程中,较多颗粒分布在连续相中而不是油水界面,因此容易发生油滴的聚合,并且如果果胶浓度太低会导致乳液内相体积比降低,体积比范围不在高内相乳液范围内,上述果胶溶液质量浓度的限定可以使乳液内相体积比保持在合适范围内。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(3)中所述大豆分离蛋白溶液与所述果胶溶液的体积比为(1:1)~(1:0.3);
进一步优选的,步骤(3)中所述大豆分离蛋白溶液与所述果胶溶液的体积比为1:1。
上述技术方案的有益效果是:确保乳液内相体积比达到高内相乳液内相体积比要求。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(4)中所述槲皮素在植物油中的浓度为0.01~0.1mg/mL;
进一步优选的,步骤(4)中所述槲皮素在植物油中的浓度为0.04mg/mL。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(4)中所述溶液体系与所述含槲皮素的植物油的体积比为(1:2)~(2:1);
进一步优选的,步骤(4)中所述溶液体系与所述含槲皮素的植物油的体积比为1:1。
上述技术方案的有益效果是:如果油相体积比太高,蛋白-果胶复合粒子不能完全包裹油滴,如果油相体积比太低,导致乳液内相体积比降低。上述溶液体系与含槲皮素的植物油的体积比可以确保油相被完全包裹。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(4)中所述超声均质乳化的功率为200~900W,间歇时间为1~5s,总时间为4~12min;
进一步优选的,步骤(4)中所述超声均质乳化的功率为600W,间歇时间为5s,总时间为6min。
上述技术方案的有益效果是:间歇时间为5s是主要由于连续超声会产生大量的热量,导致大豆分离蛋白变质,从而降低产品质量,上述条件能够确保能量消耗低,产品质量高。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(4)中所述离心的转速为8000rpm~12000rpm,离心时间为8min~15min;
进一步优选的,步骤(4)中所述离心的转速为10000rpm,离心时间为10min。
上述技术方案的有益效果是:离心后在水相顶部可以形成乳膏层,即高内相乳液。
优选的,在上述一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法中,步骤(4)中所述植物油为橄榄油、大豆油、玉米油等。
本发明还公开了一种上述方法制备得到的大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液,所述大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的平均粒径为0.5-3μm,内相体积比为79-85%,包封率为72-80%。
经由上述的技术方案可知,与现有技术相比,本发明具有以下优点和有益效果:
(1)本发明通过大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液,该凝胶状乳液内相体积比高达85%,表明乳液可有效包埋疏水性物质;
(2)本发明的乳液实现液态油向固态油的转变,没有使用任何固化剂或者表面活性添加剂;
(3)本发明中果胶的使用增加了乳液的絮凝作用,且用于稳定高内相乳液的蛋白多糖均为天然物质,具有多种优异的生理生化功能性质,绿色健康。
(4)本发明所得高内相水包油型乳液粒径小、粘弹性好,贮存稳定性高,包封率高达80%,该乳液体系在体外胃肠模拟消化中能够提高槲皮素的生物利用度,表明该乳液对活性物质具有优异的保护或控释效果;
(5)本发明的制备方法对设备要求低、方法简单易于操作,适合大规模工业化生产与加工,所得高内相水包油型乳液在化妆品、食品和药品行业具有广阔的应用空间,为药物靶向输送和功能性食品提供了一种新的思路。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案,下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍,显而易见地,下面描述中的附图仅仅是本发明的实施例,对于本领域普通技术人员来讲,在不付出创造性劳动的前提下,还可以根据提供的附图获得其他的附图。
图1-1为本发明不同pH的大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的新制外观图;
图1-2为本发明不同pH的大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的储藏1个月后外观图;
图2为实施例2大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液粒径分布图;
图3为实施例3大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液流变应力扫描图;
图4为实施例3大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液流变频率扫描图。
具体实施方式
下面将对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
实施例1
一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法,包括以下步骤:
(1)准确称取5.0g大豆分离蛋白溶解于100ml蒸馏水,得到质量浓度为5%的大豆分离蛋白溶液;将1.0g果胶溶解于100ml蒸馏水中,调节pH为2.0,得到质量浓度为1%的果胶溶液;将大豆分离蛋白溶液与果胶溶液按1:1混合;调节pH为3.0;
(2)取5.0ml步骤(1)所得溶液和含0.2mg槲皮素的5.0ml橄榄油混合,超声均质乳化的功率为570W,间歇时间为5s,时间为6min,超声后进行离心,离心的转速为10000rpm,时间为10min,得到大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液。
本实施例得到pH为3.0的O/W型高内相乳液,内相体积比达85%,制备出的乳液平均粒径为1.43±0.49μm。由图1可知,新制备的4种高内相乳液表面无漏油现象,粘稠性大,呈现凝胶状,可实现倒置,放置一个月后,pH为3的高内相乳液外观没有发生变化,无大的油滴出现,未出现破乳现象,这是由于pH值为3时果胶的凝胶效果达到最佳,其余乳液表面有黄色油滴出现,表明在较低pH值下的高内相乳液较稳定,具有良好的储藏稳定性,可放置更长时间。通过高内相乳液包埋槲皮素,包封率达80%,槲皮素的生物利用率达11.80%,为槲皮素合理剂型的开发和药理研究提供有力的证据,在新型营养物质输送载体及食品结构修饰基料应用方面具有较好的应用前景。
实施例2
一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法,包括以下步骤:
(1)准确称取5.0g大豆分离蛋白溶解于100ml蒸馏水,得到质量浓度为5%的大豆分离蛋白溶液;将1.0g果胶溶解于100ml蒸馏水中,调节pH为2.0,得到质量浓度为1%的果胶溶液;将大豆分离蛋白溶液与果胶溶液按1:1混合;调节pH为7.0;
(2)取5.0ml步骤(1)所得溶液和含0.2mg槲皮素的5.0ml橄榄油混合,超声均质乳化的功率为550W,间歇时间为5s,时间为8min。超声后进行离心,离心的转速为10000rpm,时间为8min,得到大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液。
采用Image J方法测量高内相乳液的直径,pH为7.0的乳液粒径分布如图2所示。
本实施例得到pH为7.0的O/W型高内相乳液,内相体积比达81%,包封率达77%,经体外模拟消化槲皮素的生物利用率达15.49%。制备得到的高内相乳液外观呈现均匀统一,倒置不发生掉落,放置一个月后,高内相乳液表面有黄色油滴出现,开始出现溢油现象。图2表明,pH为7.0的大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液在新制时粒径大小一致,均匀分布在0.5um~3um范围内。乳液粒径越小,则表明乳液越稳定,说明本发明制备的高内相乳液有望稳定荷载易被降解的活性物质构建一套符合人体消化吸收特点的输送体系,提高活性物质的生物利用率,达到缓控释的效果。
实施例3
一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法,包括以下步骤:
(1)准确称取5.0g大豆分离蛋白溶解于100ml蒸馏水,得到质量浓度为5%的大豆分离蛋白溶液;将1.0g果胶溶解于100ml蒸馏水中,调节pH为2.0,得到质量浓度为1%的果胶溶液;将大豆分离蛋白溶液与果胶溶液按1:1混合;调节pH为9.0;
(2)取5.0ml步骤(1)所得溶液和含0.2mg槲皮素的5.0ml橄榄油混合,超声均质乳化的功率为600W,间歇时间为5s,时间为10min。超声后进行离心,离心的转速为10000rpm,时间为10min,得到大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液。
取制得的乳液2ml进行流变学特性测试,使用直径为30mm的平板,温度为25℃,应力扫描:设置应力范围为0.1Pa到1000Pa,固定频率为1Hz,其流变学特性图如图3所示。频率扫描:固定应力为1Pa,频率范围为0.1Hz到10Hz,其流变学特性如图4所示。
本实施例得到pH为9.0的O/W型高内相乳液,内相体积比达79%,包封率达72%。制备得到的高内相乳液外观呈现均匀统一的乳白色,可以实现倒置,乳液形成了凝胶状网络结构,从图3、4可知,乳液表现出凝胶特性(弹性模量远大于粘性模量),并且随着应力的增大,屈服点也随之出现,具有较宽的线性粘弹区域。该高内相乳液的应力扫描弹性模量超过2000Pa,在压力增大到150Pa时才出现临界交点,频率扫描弹性模量也超过2000Pa,而在流变学特性实验中,一般高内相乳液的应力扫描弹性模量在1000Pa左右,频率扫描的弹性模量也在1000Pa左右,这表明通过超声法制备的高内相乳液具有优异的稳定性、粘弹性和凝胶性。
本发明通过超声波均质技术,蛋白-多糖胶体粒子可同时稳定多个油滴,使得油滴与油滴之间被胶体粒子固定下来,失去流动性,形成了更为固定的三维网络结构,达到较强的凝胶强度。乳液实现了液态油脂向固态油脂的转变,可取代具有反式酸的固态油脂,在食品、药品和化妆品领域有较好的应用前景。
本说明书中各个实施例采用递进的方式描述,每个实施例重点说明的都是与其他实施例的不同之处,各个实施例之间相同相似部分互相参见即可。对于实施例公开的装置而言,由于其与实施例公开的方法相对应,所以描述的比较简单,相关之处参见方法部分说明即可。
对所公开的实施例的上述说明,使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下,在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
Claims (2)
1.一种大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的制备方法,其特征在于,包括以下步骤:
(1)将大豆分离蛋白溶于蒸馏水中,获得质量浓度为5.0%的大豆分离蛋白溶液;
(2)将果胶溶于蒸馏水中,调节pH为2.0~4.0,获得质量浓度为1%的果胶溶液;
(3)将所述大豆分离蛋白溶液与所述果胶溶液按体积比为1:1混合,调节混合溶液的pH为3.0~11.0,得到溶液体系;
(4)将所述溶液体系与含槲皮素的植物油混合,进行超声均质乳化,离心后获得大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液;
所述槲皮素在植物油中的浓度为0.04mg/mL,所述溶液体系与所述含槲皮素的植物油的体积比为1:1;所述超声均质乳化的功率为200~900W,间歇时间为5s,总时间为4~12min;所述离心的转速为10000rpm,离心时间为8min~15min。
2.一种权利要求1所述方法制备得到的大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液,其特征在于,所述大豆分离蛋白-果胶复合物稳定槲皮素的高内相乳液的粒径为0.5-3μm,内相体积比为79-85%,包封率为72-80%。
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