CN114432263A - 一种壳聚糖修饰的叶黄素纳米结构脂质载体及其制备方法 - Google Patents
一种壳聚糖修饰的叶黄素纳米结构脂质载体及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种壳聚糖修饰的叶黄素纳米结构脂质载体及其制备方法,通过小分子表面活性剂吐温‑80与天然生物大分子乳清分离蛋白(Whey protein isolate,WPI)进行复配,构建复合界面的叶黄素纳米结构脂质载体,再通过壳聚糖对其进行二次界面修饰,获得壳聚糖修饰的叶黄素纳米结构脂质载体,进一步增强了叶黄素对光、热和胃肠道环境的耐受性,在胃肠液环境中,经壳聚糖二次修饰后的叶黄素纳米结构脂质载体具有良好的控释行为,提高了叶黄素生物可给率,可有效避免常规乳液等运载体系在消化过程中出现的脂肪快速消化和活性物质“突释”的问题。
Description
技术领域
本发明涉及食品加工技术领域,尤其是一种壳聚糖修饰的叶黄素纳米结构脂质载体及其制备方法。
背景技术
我国目前面临着非常严峻的用眼健康问题,截至2020年12月,我国手机网民规模达9.86亿,视疲劳、白内障和老年黄斑变性病等眼部疾病在不同年龄人群中越发增多。叶黄素(Lutein)是一种脂溶性类胡萝卜素,具有较强的抗氧化能力,可以特异性积聚在黄斑区,降低湿性年龄相关性黄斑变性,可保护眼睛免受高能量光线伤害,阻止眼晶状体氧化、降低白内障风险等多种生理功能。然而,叶黄素分子含有多个共轭双键,稳定性差、水溶性低,易分解、聚集、结晶析出而无法与胆汁盐形成叶黄素混合胶束,降低了在人体的吸收利用率,这在很大程度上限制了叶黄素作为一种功能性营养素在食品和医药等领域的应用。叶黄素的生物可给率指的是膳食中叶黄素摄入后被载入混合胶束中成为可被小肠吸收的比例。因此,促进叶黄素在小肠中形成混合胶束是提高其生物可及性的关键。研究发现,构建合适的纳米脂质递送载体可有效改善叶黄素生物可及性并起到缓释作用。
纳米结构脂质载体(Nanostructured lipid carriers,NLCs)作为新一代脂质纳米粒子,因具有突出的控释效果,可增加脂溶性药物的作用时间,提高生物可给率等优点,成为活性成分递送体系领域的研究热点。负载叶黄素的NLCs摄入到人体进入肠相后,胆盐首先吸附在油水界面上发生界面取代,随后脂肪酶吸附在脂滴界面上来水解脂质,同时释放类胡萝卜素,并与胆盐、FFAs、MAGs混合形成胶束离开油水界面层,再进一步被人体吸收。胰脂肪酶是一种界面酶,需要吸附在油水界面上才能发挥催化作用,在消化过程中,NLCs界面层是最先接触胃肠道环境(低pH、消化酶和金属离子)的部分,因此,胆盐在油水界面对表面活性剂的取代作用影响脂肪酶对油脂的消化水解,而胆盐的取代作用又取决于界面性质。表面活性剂是构成NLCs界面的主要成分,它可以降低互不相容两相间的界面张力,起到稳定乳液的作用。
目前,针对脂质运载体系,国内外研究热点主要集中在设计载体的精细界面结构,来调控其稳定性和消化特性,由小分子表面活性剂形成的单界面层存在界面层较薄的缺陷,极易被胆盐和胰脂肪酶取代,从而载体脂质容易被脂肪酶快速水解。研究发现,利用生物大分子蛋白或多糖对界面进行修饰可提高界面层致密度和厚度,增加界面阻隔性能,进而抑制胆盐、胰脂肪酶以及Ca2+的界面扩散,调控脂质运载体系中活性物质的释放。通常,研究者多数采用小分子表面活性剂作为单一乳化剂构建单界面层NLCs,近年的研究表明,通过天然大分子比如蛋白质类、多糖类等对NLCs界面层进行适当修饰,可以有效改善脂溶性活性物质的稳定性、缓释效果以及生物可给率,其中壳聚糖及其衍生物应用最为广泛。
发明内容
本发明要解决的技术问题是:针对常规技术制备的叶黄素递送体系存在对热和酸敏感,粒径变大、絮凝、芯材渗漏等不稳定问题,本技术提供一种壳聚糖修饰的叶黄素纳米结构脂质载体及其制备方法,旨在提高叶黄素对光、热和胃肠道环境的耐受性,提高其稳定性,增强叶黄素在结肠中发挥更长效的释放。
本发明解决其技术问题所采用的技术方案是:一种壳聚糖修饰的叶黄素纳米结构脂质载体包括以下重量份的组分:叶黄素0.2~1份,亚麻籽油和山嵛酸甘油酯混合相19~20份,吐温-80溶液0.2~0.4份,WPI溶液0.2~0.4份,水50~70份,壳聚糖稀酸溶液10~30份,其制备方法采用高压微射流法制备法,具体操作步骤如下:
步骤(1),将亚麻籽油和山嵛酸甘油酯按照一定的比例混合后,80℃水浴加热,搅拌直至完全熔化;
步骤(2),将一定质量的叶黄素加入到步骤(1)所述的混合油相,通过磁力搅拌使其充分溶解在混合油相中;
步骤(3),将一定浓度的WPI溶液与1%的吐温-80溶液等比例混合,预热到80℃后迅速加入到步骤(2)所述的混合油相中,搅拌形成分散液;
步骤(4),将步骤(3)所述的分散液通过高速剪切一定时间形成预乳液,再通过高压微射流均质处理后,得到预乳液;
步骤(5),精确称取壳聚糖粉末溶于稀酸溶液中,在50℃加热搅拌8h,过滤取出少量不溶物质;
步骤(6),将步骤(4)所述的预乳液置于烧杯中,室温搅拌,匀速滴加一定质量步骤(5)所述的壳聚糖稀酸溶液,继续搅拌5~8h,迅速冷却得到壳聚糖修饰的叶黄素纳米结构脂质载体,充入N2密封,置于冰箱冷藏保存。
作为优选的技术方案,步骤(1)中,亚麻籽油和山嵛酸甘油酯质量比为3:7。
作为优选的技术方案,步骤(2)中,叶黄素占混合油相的质量分数为1%~5%。
作为优选的技术方案,步骤(3)中,WPI溶液的浓度为1%~4%。
作为优选的技术方案,步骤(3)中,混合油相粘20%(w/w),吐温-80溶液与WPI溶液占80%(w/w)。
作为优选的技术方案,步骤(4)中,高速剪切转速为14000r/min,搅拌时间为20min;高压微射流压力为20400psi,循环次数3次。
作为优选的技术方案,步骤(5)中,壳聚糖稀酸溶液的浓度为0.5mg/L~3mg/L,壳聚糖稀酸溶液占预乳液的质量分数为10%~30%。
作为优选的技术方案,步骤(5)中,稀酸溶液包括75mmol/LNaCl、0.01mol/L磷酸盐、0.1mol/L醋酸。
本发明的一种壳聚糖修饰的叶黄素纳米结构脂质载体及其制备方法,其有益效果是:
1.本发明制备的壳聚糖修饰的叶黄素纳米结构脂质载体,材料组成简单,安全可靠,有效增强了叶黄素对光、热和胃肠道环境的耐受性,提高了稳定性,有效降低释放速率,解决了常规纳米乳液体系易“突释”的问题,显著提高了叶黄素生物可给率。
2.本发明制备的壳聚糖修饰的叶黄素纳米结构脂质载体粒径为150nm~320nm,叶黄素加热后保留率为64.6%~75.8%,光照条件下叶黄素的保留率达65.3%~77.4%,释放时间为12~24h,叶黄素生物可给率为53.7%~65.9%。
附图说明
下面结合附图和实施例对本发明进一步说明。
图1为本发明的壳聚糖修饰的叶黄素纳米结构脂质载体的透射电镜图;
图2为本发明的壳聚糖修饰的叶黄素纳米结构脂质载体加热条件下叶黄素保留率图。
具体实施方式
现在实施例对本发明作进一步详细的说明。
一种壳聚糖修饰的叶黄素纳米结构脂质载体,包括以下重量份的组分:叶黄素0.2~1份,亚麻籽油和山嵛酸甘油酯混合相19~19.8份,吐温-80溶液0.28~0.36份,WPI溶液0.28~0.36份,水55.44~71.28份,壳聚糖稀酸溶液10~30份。
一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,具体包括以下步骤:
步骤(1),将亚麻籽油和山嵛酸甘油酯按照一定的比例混合后,80℃水浴加热,搅拌直至完全熔化;
步骤(2),将一定质量的叶黄素加入到步骤(1)所述的混合油相,通过磁力搅拌使其充分溶解在混合油相中;
步骤(3),将一定浓度的WPI溶液与1%的吐温-80溶液等比例混合,预热到80℃后迅速加入到步骤(2)所述的混合油相中,搅拌形成分散液;
步骤(4),将步骤(3)所述的分散液通过高速剪切一定时间形成预乳液,再通过高压微射流均质处理后,得到预乳液;
步骤(5),精确称取壳聚糖粉末溶于稀酸溶液中,在50℃加热搅拌8h,过滤取出少量不溶物质;
步骤(6),将步骤(4)所述的预乳液置于烧杯中,室温搅拌,匀速滴加一定质量步骤(5)所述的壳聚糖稀酸溶液,继续搅拌5~8h,迅速冷却得到壳聚糖修饰的叶黄素纳米结构脂质载体,充入N2密封,置于冰箱冷藏保存。
实施例1
将亚麻籽油和山嵛酸甘油酯按照30%:70%(w/w)的比例混合后,于80℃条件下加热熔化,然后加入叶黄素(占总油脂的质量分数为2%),通过磁力搅拌使其充分溶解在混合油相中。将加热到80℃的1%吐温-80的水溶液和2%WPI溶液等比例混合迅速加入到混合油相中,其中,混合油脂(油相)占20%(w/w),吐温-80水溶液和WPI溶液(水相)占80%(w/w),在1400r/min下搅拌20min,然后经20400psi高压微射流,循环处理3次后,得到预乳液。将预乳液置于烧杯中,室温搅拌,匀速滴加一定质量的0.5mg/L的壳聚糖稀酸溶液,预乳液与壳聚糖稀酸溶液的质量比为9:1,继续搅拌5~8h,迅速冷却得到壳聚糖修饰的叶黄素纳米结构脂质载体,充入N2密封,置于冰箱冷藏保存。经测定,壳聚糖修饰的叶黄素纳米结构脂质载体粒径为162nm,叶黄素加热后保留率为66.5%,光照条件下叶黄素的保留率达69.6%,释放时间为15h,叶黄素生物可给率为59.4%。
实施例2
将亚麻籽油和山嵛酸甘油酯按照30%:70%(w/w)的比例混合后,于80℃条件下加热熔化,然后加入叶黄素(占总油脂的质量分数为2%),通过磁力搅拌使其充分溶解在混合油相中。将加热到80℃的1%吐温-80的水溶液和3%WPI溶液等比例混合迅速加入到混合油相中,其中,混合油脂(油相)占20%(w/w),吐温-80水溶液和WPI溶液(水相)占80%(w/w),在1400r/min下搅拌20min,然后经20400psi高压微射流,循环处理3次后,得到预乳液。将预乳液置于烧杯中,室温搅拌,匀速滴加一定质量的1mg/L的壳聚糖稀酸溶液,预乳液与壳聚糖稀酸溶液的质量比为8:2,继续搅拌5~8h,迅速冷却得到壳聚糖修饰的叶黄素纳米结构脂质载体,充入N2密封,置于冰箱冷藏保存。经测定,壳聚糖修饰的叶黄素纳米结构脂质载体粒径为224nm,叶黄素加热后保留率为72.6%,光照条件下叶黄素的保留率达74.3%,释放时间为14h,叶黄素生物可给率为61.8%。
实施例3
将亚麻籽油和山嵛酸甘油酯按照30%:70%(w/w)的比例混合后,于80℃条件下加热熔化,然后加入叶黄素(占总油脂的质量分数为2%),通过磁力搅拌使其充分溶解在混合油相中。将加热到80℃的1%吐温-80的水溶液和2%WPI溶液等比例混合迅速加入到混合油相中,其中,混合油脂(油相)占20%(w/w),吐温-80水溶液和WPI溶液(水相)占80%(w/w),在1400r/min下搅拌20min,然后经20400psi高压微射流,循环处理3次后,得到预乳液。将预乳液置于烧杯中,室温搅拌,匀速滴加一定质量的2.5mg/L的壳聚糖稀酸溶液,预乳液与壳聚糖稀酸溶液的质量比为7:3,继续搅拌5~8h,迅速冷却得到壳聚糖修饰的叶黄素纳米结构脂质载体,充入N2密封,置于冰箱冷藏保存。经测定,壳聚糖修饰的叶黄素纳米结构脂质载体粒径为242nm,叶黄素加热后保留率为71.4%,光照条件下叶黄素的保留率达76.5%,释放时间为21h,叶黄素生物可给率为62.4%。
由图1的透射电镜图看出,本发明制备的壳聚糖修饰的叶黄素纳米结构脂质载体,粒度圆整,分散较好;由图2的叶黄素保留率图可知,本发明制备的壳聚糖修饰的叶黄素纳米结构脂质载体在加热条件下其叶黄素保留率得到明显提升。
本发明制备的壳聚糖修饰的叶黄素纳米结构脂质载体粒径为150nm~320nm,叶黄素加热后保留率达64.6%~75.8%,光照条件下叶黄素的保留率达65.3%~77.4%,释放时间为12~24h,叶黄素生物可给率达53.7%~65.9%。
本发明通过小分子表面活性剂吐温-80与天然生物大分子乳清分离蛋白(Wheyprotein isolate,WPI)进行复配,构建复合界面的叶黄素纳米结构脂质载体,再通过壳聚糖对其进行二次界面修饰,获得壳聚糖修饰的叶黄素纳米结构脂质载体,并进一步增强了叶黄素对光、热和胃肠道环境的耐受性,在胃肠液环境中,经壳聚糖二次修饰后的叶黄素纳米结构脂质载体具有良好的控释行为,提高了叶黄素生物可给率,可有效避免常规乳液等运载体系在消化过程中出现的脂肪快速消化和活性物质“突释”的问题。
以上述依据本发明的理想实施例为启示,通过上述的说明内容,相关工作人员完全可以在不偏离本项发明技术思想的范围内,进行多样的变更以及修改。本项发明的技术性范围并不局限于说明书上的内容,必须要根据权利要求范围来确定其技术性范围。
Claims (9)
1.一种壳聚糖修饰的叶黄素纳米结构脂质载体,其特征是:包括以下重量份的组分:叶黄素0.2~1份,亚麻籽油和山嵛酸甘油酯混合相19~20份,吐温-80溶液0.2~0.4份,WPI溶液0.2~0.4份,水50~80份,壳聚糖稀酸溶液10~30份。
2.根据权利要求1所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:具体包括以下步骤:
步骤(1),将亚麻籽油和山嵛酸甘油酯按照一定的比例混合后,80℃水浴加热,搅拌直至完全熔化;
步骤(2),将一定质量的叶黄素加入到步骤(1)所述的混合油相,通过磁力搅拌使其充分溶解在混合油相中;
步骤(3),将一定浓度的WPI溶液与1%的吐温-80溶液等比例混合,预热到80℃后迅速加入到步骤(2)所述的混合油相中,搅拌形成分散液;
步骤(4),将步骤(3)所述的分散液通过高速剪切一定时间形成预乳液,再通过高压微射流均质处理后,得到预乳液;
步骤(5),精确称取壳聚糖粉末溶于稀酸溶液中,在50℃加热搅拌8h,过滤取出少量不溶物质;
步骤(6),将步骤(4)所述的预乳液置于烧杯中,室温搅拌,匀速滴加一定质量步骤(5)所述的壳聚糖稀酸溶液,继续搅拌5~8h,迅速冷却得到壳聚糖修饰的叶黄素纳米结构脂质载体,充入N2密封,置于冰箱冷藏保存。
3.根据权利要求2所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:步骤(1)中,亚麻籽油和山嵛酸甘油酯质量比为3:7。
4.根据权利要求2所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:步骤(2)中,叶黄素占混合油相的质量分数为1%~5%。
5.根据权利要求2所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:步骤(3)中,WPI溶液的浓度为1%~4%。
6.根据权利要求2所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:步骤(3)中,混合油相粘20%(w/w),吐温-80溶液与WPI溶液占80%(w/w)。
7.根据权利要求2所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:步骤(4)中,高速剪切转速为14000r/min,搅拌时间为20min;高压微射流压力为20400psi,循环次数3次。
8.根据权利要求2所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:步骤(5)中,壳聚糖稀酸溶液的浓度为0.5mg/L~3mg/L,壳聚糖稀酸溶液占预乳液的质量分数为10%~30%。
9.根据权利要求2所述的一种壳聚糖修饰的叶黄素纳米结构脂质载体的制备方法,其特征是:步骤(5)中,稀酸溶液包括75mmol/LNaCl、0.01mol/L磷酸盐、0.1mol/L醋酸。
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