CN117257675A - 一种环状γ-聚谷氨酸纳米微胶囊及其制备方法与应用 - Google Patents
一种环状γ-聚谷氨酸纳米微胶囊及其制备方法与应用 Download PDFInfo
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Abstract
本发明涉及一种环状γ‑聚谷氨酸纳米微胶囊及其制备方法与应用。步骤如下:a:制备芯材溶液、环状γ‑聚谷氨酸水溶液和壳聚糖溶液,所述芯材为苯乙基间苯二酚、对羟基苯乙酮、白藜芦醇、光甘草定或虾青素;b:将芯材溶液滴加至环状γ‑聚谷氨酸水溶液中,搅拌,得到浓缩溶液;c:将浓缩溶液滴加至壳聚糖溶液中,搅拌,经过滤、冷冻、干燥后,得环状γ‑聚谷氨酸纳米微胶囊。本发明提供的环状γ‑聚谷氨酸纳米微胶囊是以环状γ‑聚谷氨酸和壳聚糖为材料对芯材进行包裹得到,具有包封率高,缓释效果好的优点,并且原料使用明确,操作简单,条件温和,适合大规模工业化生产。
Description
技术领域
本发明涉及一种环状γ-聚谷氨酸纳米微胶囊及其制备方法与应用,属于日用护肤品技术领域。
背景技术
环状γ-聚谷氨酸分子量小,没有游离的N端和C端,对酶的敏感性降低,稳定性提高,其四周具有水合能力的官能团及氢键,环中间为疏水结构,能增加非水溶性物质的亲水性及稳定性,并可以与带正电的有效成分发生吸引,创造一个优良的包裹、缓释系统,让化妆品内的有效成分可以慢慢发挥其功能,使皮肤更有效率地吸收保养品中各种功效成分。环γ-聚谷氨酸具有抑制酪氨酸酶活性的功能,可以减少皮肤黑色素产生。此外,环γ-聚谷氨酸可以被细胞表面的γ-谷氨酰转肽酶特异识别,并通过其介导的内吞途径进入细胞,具备大分子量γ-聚谷氨酸不具有的进入皮肤细胞能力。
γ-聚谷氨酸用于载体需要经过修饰或者与目标药品结合,工艺复杂。环状γ-聚谷氨酸通过包裹产物可以避免这个问题,对于低分子量γ-聚谷氨酸的研究,主要集中在医药载体领域,环γ-聚谷氨酸在化妆品领域的研究则还较少见报导,特别是其助脂溶性物质溶解功能未见报导。中国专利文献CN108610274A从水母刺丝囊提取环γ-聚谷氨酸,用于医药领域的蛋白质制剂分子载体。中国专利文献CN116199641A通过化学法合成环γ-聚谷氨酸,但没有具体使用方案。
苯乙基间苯二酚、对羟基苯乙酮、白藜芦醇、光甘草定和虾青素均是日用护肤品中常用的成分。其中苯乙基间苯二酚是一种可以抑制酪氨酸酶,具有抗氧化作用,降低黑色素生成的美白活性成份。白藜芦醇具有显著的抗氧化功能。虾青素能吸收紫外线,减少对皮肤的伤害,有很强的抗氧化,清除自由基能力,能保护皮肤细胞的DNA从而减少黑色素合成,具有美白、淡斑、抗衰老、抗炎、抗感染等功效。但虾青素具有脂溶性特征,不溶于水,,同时易受光、热、氧等影响而分解,限制其实际使用,研究人员提高虾青素利用率的技术有纳米乳化、微胶囊法、包埋法。化妆用化妆品主要分为油包水型和水包油型,水包油型化妆品持久力强,但厚重黏腻,油包水型化妆品轻爽,但持久力差。为弥补目前化妆品组合物的缺点,利用环γ-聚谷氨酸特性,结合虾青素和壳聚糖特点,提高皮肤补水美白能力。环γ-聚谷氨酸有助于油性溶剂溶解,将化妆品中油性成分包裹后分散,形成均一分散体系,使化妆品清爽且持久,缓慢释放,减少其他乳化剂使用。
目前专利及文献中,为了提高虾青素水溶性及利用主要技术有纳米乳化、微胶囊及包埋等,都存在一定问题。中国专利CN110269254A提供一种美白、抗氧化、增强免疫力的纳乳及其制备方法和应用,将雨生红球藻粉在动植物油中溶解后加入乳化剂,高压匀质后得到纳米乳液,具有操作简单、稳定性好、方便食用、吸收率高等优点,但其主要用于食用,不适合做护肤品,且吸收过快,体内作用时间短,不具备缓释功效,此外使用了额外的油脂和乳化剂。中国专利CN116211746A提供一种雨生红球藻源虾青素水乳剂的制备方法,用雨生红球藻油加入油脂、抗氧化剂、乳化剂等高压均质得到虾青素水乳剂,具有载量高,溶解性高且稳定的优点,但大量使用油脂及乳化剂维持状态,使用时厚重黏腻。
中国专利CN109646425A提供一种H1、H2或J型虾青素聚集体水分散体系的制备方法与应用,将富含虾青素用乙醇萃取后,混合甲壳素及DNA溶液,最终得到水分散体系胶体或固体粉末。利用天然生物大分子壳聚糖、鱼精DNA构成的疏水空间维持虾青素的稳定性及水溶性,未添加其他油剂、表面活性剂的化学合成物质,条件温和,生产成本低、绿色无污染。但鱼精DNA提取成本较高,纯度及生物相容性存在一些不足,不利于大规模放大。中国专利CN111793014A提供一种制备水溶性虾青素的方法及由其制得的虾青素水溶液,将富含虾青素的原料与含有机酸溶液混合,破碎后多次浸取,实现天然虾青素与原料中天然蛋白质、核酸、多糖等作用,一步得到水溶性虾青素,来源天然,操作简单,成本低,产物无有机溶剂残留。但其利用原料中的蛋白质、核酸、多糖等成分不明确,还会有很多水溶性杂质,可能存在潜在过敏隐患。
中国专利CN108403666A提供一种新型虾青素纳米微粒的制备方法,将虾青素和PLGA的丙酮溶液加入含乳化剂TPGS的水溶液,乳化均质得到纳米乳剂,旋蒸后得到纳米微球,具有缓释、稳定、包埋率高等优点,但是其使用丙酮等有毒有机溶剂和乳化剂,存在残留风险,生产操作不连续等问题。中国专利CN104784059A提供一种类胡萝卜素包合物和聚谷氨酸组合物及其用途,该专利使用环糊精包埋,利用其内部疏水空间增加虾青素水溶性,γ-聚谷氨酸单纯提供补水能力,让皮肤湿润。但环糊精及γ-聚谷氨酸都不具备穿透皮肤的能力,会被皮肤角质层阻碍,其通过环糊精增加的水溶性并不能被细胞吸收,但其能使包含物渗透系数增大,通过缓慢渗透作用进入人体。此外γ-聚谷氨酸用量过大,为虾青素的400倍,会使产品成膜有黏腻感,实际使用及吸收效果较差,并没有真正提高水溶性。在“Thepreparation and stability of the inclusion complex of astaxanthin withβ-cyclodextrin”中,便有将虾青素包合在β-环糊精的方法,但载药量低,造成资源浪费。
γ-聚谷氨酸做载体通常需要利用聚乙二醇、苄胺等对其修饰,或者将有效药用成分与其结合,化学法制备复合物,工艺复杂且容易残留有机溶剂,γ-聚谷氨酸与壳聚糖自组装纳米颗粒来装载药物,在一些专利中也有应用,也有很多问题,先制备成纳米微胶囊会影响药物吸附结合,载药效果差。
在表面活性剂方面,随着生活水平提高及市场发展,消费者对化妆品有着高品质、高安全性和环保性的需求,逐渐淘汰有害、有毒类表面活性剂,趋向于使用绿色、功能化发展。中国专利CN11616978A提供油菜油酰胺丙基二甲胺作为乳化剂在化妆品中的应用,提供一种产物的新用途,除了可以作为乳化剂,还可以抑制微生物生长,避免硅油添加,但化妆品偏酸性限制与其他成分混合使用,配伍性不高。中国专利CN106890097A提供了一种水包油型颗粒乳化剂和化妆品的制备,利用改性的透明质酸作为乳化剂,通过自组装构成胶体粒子,在具有乳化效果的同时还具有保湿性及吸收紫外的特点,不过油相油脂及助乳化剂用量大,使用会有厚重感,不清爽。
发明内容
针对现有技术的不足,本发明提供了一种环状γ-聚谷氨酸纳米微胶囊及其制备方法与应用。
本发明的技术方案如下:
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,包括步骤如下:
步骤a:在避光条件下,将芯材溶于溶剂中,得到芯材溶液;将环状γ-聚谷氨酸溶于水中,得到环状γ-聚谷氨酸水溶液;将壳聚糖溶于醋酸中,得到壳聚糖溶液;
所述芯材为苯乙基间苯二酚、对羟基苯乙酮、白藜芦醇、光甘草定或虾青素;
所述溶剂为无水乙醇或水;
步骤b:一边搅拌,一边将芯材溶液滴加至环状γ-聚谷氨酸水溶液中,滴加完成后继续搅拌1~2h后,除去乙醇,得到浓缩溶液;
步骤c:一边搅拌,一边将浓缩溶液滴加至壳聚糖溶液中,滴加完成后继续搅拌5~10min,经过滤、冷冻、干燥后,得到环状γ-聚谷氨酸纳米微胶囊。
根据本发明优选的,步骤a中,所述芯材溶液的浓度为0.02~1mg/mL;所述芯材为虾青素。
根据本发明优选的,步骤a中,所述环状γ-聚谷氨酸溶液的浓度为0.2~5mg/mL。
根据本发明优选的,步骤a中,所述壳聚糖溶液的浓度为1~5mg/mL。
根据本发明优选的,步骤b中,所述芯材溶液和环状γ-聚谷氨酸水溶液的体积比为(1~5):(5~15)。
根据本发明优选的,步骤b中,所述搅拌速度为200~1000r/min,滴加速度为50~100mL/min。
根据本发明优选的,步骤b中,通过旋转蒸发仪除去乙醇,旋蒸条件为30~40℃,真空度为1~5mbar。
根据本发明优选的,步骤c中,所述搅拌速度为1500~2000r/min,滴加速度为10~50mL/min。
本发明还提供了一种环状γ-聚谷氨酸纳米微胶囊,由上述技术方案中所述的制备方法制得。
根据本发明优选的,所述环状γ-聚谷氨酸纳米微胶囊的粒度范围是100~200nm。
本发明还提供了上述环状γ-聚谷氨酸纳米微胶囊在制备化妆品、医美产品中的应用。
有益效果:
1、本发明提供的环状γ-聚谷氨酸纳米微胶囊是以环状γ-聚谷氨酸和壳聚糖为材料对芯材进行包裹得到,具有包封率高,缓释效果好的优点。环状γ-聚谷氨酸(分子量为200~1000Da)可以快速被皮肤吸收,环状γ-聚谷氨酸可以通过γ-谷氨酰转肽酶介导的细胞内吞途径进入细胞,特别对虾青素水溶性的增强可以确保其被细胞吸收并发挥作用,无需额外添加大量脂质来促进吸收,制备的产品水溶后清爽无黏腻感。并且环状γ-聚谷氨酸具有在水中溶解分散的功能,其与虾青素分子量相当,可以在水中自发组装成纳米级胶束,具有更小的纳米结构便于吸收,避免高压高速搅拌均质。油溶性虾青素经环状γ-聚谷氨酸包裹后易于分散,有效地解决了虾青素溶解性差、吸收性差的问题,并且包裹后有助于防止虾青素氧化。同时壳聚糖带有正电荷,与环状γ-聚谷氨酸有良好的吸附效果,也具有保湿、美白、抗炎功效,与环状γ-聚谷氨酸包裹虾青素后具有良好的缓释功能。
2、本发明提供的环状γ-聚谷氨酸纳米微胶囊制备方法不需要使用过多油剂、表面活性剂、抗氧化剂,得到的环状γ-聚谷氨酸纳米微胶囊具有良好的生物安全性、可降解性。
3、本发明提供的环状γ-聚谷氨酸纳米微胶囊制备方法原料使用明确,操作简单,条件温和,无需高压均质,适合大规模工业化生产。
4、本发明提供的环状γ-聚谷氨酸纳米微胶囊中,环状γ-聚谷氨酸成环后形成亲水及疏水空间,可以增加非水溶性物质的亲水性及稳定性,并可以与带正电的有效成分发生吸引能力,创造一个优良的包裹、缓释系统。特别是虾青素提取后以单酯或双酯的形式存在,难溶于水,通过环状γ-聚谷氨酸包裹后增加其溶解性,得到透明溶液,并通过包裹来减少虾青素的氧化分解,增加稳定性。壳聚糖是自然界中唯一携带正电荷的多糖,可以与环状γ-聚谷氨酸稳定静电吸附,进一步增强γ-聚谷氨酸包裹虾青素后的靶向性及缓释功效,避免虾青素吸收后在体内迅速代谢、消耗,增加功效持久性。实现了一种物质多种功效,不同组分根据特性筛选能有效结合,超出各自的原本功效。并避免了油脂及一些乳化剂的使用,通过环状γ-聚谷氨酸包裹也可避免其他抗氧化剂的添加。产品清爽持久,具有美白、保湿、抗氧化等多种功效,可以广泛应用于食品、化妆品。
具体实施方式
下面将结合本发明实施例,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
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,包括步骤如下:
步骤a:在避光条件下,将50mg虾青素溶于500mL无水乙醇中,得到浓度为0.1mg/mL的虾青素溶液;将200mg环状γ-聚谷氨酸溶于200mL水中,得到浓度为1mg/mL的环状γ-聚谷氨酸水溶液;将500mg壳聚糖溶于500mL醋酸(1wt%)中,得到浓度为1mg/mL的壳聚糖溶液;
步骤b:在500r/min的搅拌速度下,按照60mL/min速率将500mL虾青素溶液滴加至200mL环状γ-聚谷氨酸水溶液中,滴加完成后继续搅拌1.5h后,除去乙醇,得到浓缩溶液;
步骤c:在1800r/min的搅拌速度下,按照30mL/min速率将150mL浓缩溶液滴加至500mL壳聚糖溶液中,滴加完成后继续搅拌8min,经0.45μm微孔滤膜过滤后,冷冻干燥机冷冻干燥后,得到环状γ-聚谷氨酸纳米微胶囊。
实施例2
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,包括步骤如下:
步骤a:在避光条件下,将10mg虾青素溶于500mL无水乙醇中,得到浓度为0.02mg/mL的虾青素溶液;将40mg环状γ-聚谷氨酸溶于200mL水中,得到浓度为0.2mg/mL的环状γ-聚谷氨酸水溶液;将500mg壳聚糖溶于400mL醋酸(1wt%)中,得到浓度为1.25mg/mL的壳聚糖溶液;
步骤b:在200r/min的搅拌速度下,按照80mL/min将500mL虾青素溶液滴加至200mL环状γ-聚谷氨酸水溶液中,滴加完成后继续搅拌1h后,除去乙醇,得到浓缩溶液;
步骤c:在1500r/min的搅拌速度下,按照20mL/min将120mL浓缩溶液滴加至400mL壳聚糖溶液中,滴加完成后继续搅拌5min,经0.45μm微孔滤膜过滤后,冷冻干燥机冷冻干燥后,得到环状γ-聚谷氨酸纳米微胶囊。
实施例3
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,包括步骤如下:
步骤a:在避光条件下,将500mg虾青素溶于500mL无水乙醇中,得到浓度为1mg/mL的虾青素溶液;将1000mg环状γ-聚谷氨酸溶于200mL水中,得到浓度为5mg/mL的环状γ-聚谷氨酸水溶液;将5000mg壳聚糖溶于1000mL醋酸(1wt%)中,得到浓度为5mg/mL的壳聚糖溶液;
步骤b:在1000r/min的搅拌速度下,按照100mL/min将500mL虾青素溶液滴加至200mL环状γ-聚谷氨酸水溶液中,滴加完成后继续搅拌2h后,除去乙醇,得到浓缩溶液;
步骤c:在2000r/min的搅拌速度下,按照30mL/min将180mL浓缩溶液滴加至1000mL壳聚糖溶液中,滴加完成后继续搅拌10min,经0.45μm微孔滤膜过滤后,冷冻干燥机冷冻干燥后,得到环状γ-聚谷氨酸纳米微胶囊。
实施例4
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,步骤同实施例1所述不同之处在于,芯材为苯乙基间苯二酚。
实施例5
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,步骤同实施例1所述不同之处在于,芯材为白藜芦醇。
实施例6
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,步骤同实施例1所述不同之处在于,芯材为光甘草定。
实施例7
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,步骤同实施例1所述不同之处在于,芯材为对羟基苯乙酮,将对羟基苯乙酮溶于水中,温度为55℃。
对比例1
一种环状γ-聚谷氨酸纳米微胶囊的制备方法,步骤如实施例1所述,不同之处在于,使用γ-聚谷氨酸代替环状γ-聚谷氨酸。
对比例2
按照中国专利文献CN102058562A公开的方法制备纳米胶囊,然后用该纳米胶囊包埋虾青素。
实验例
1、包封率测定
分别准确称取50mg实施例1~7和对比例1~2制备的纳米微胶囊样品于1.5mL离心管,然后加入1mL正己烷,在室温条件下15000r/min条件下离心10min,过滤,测定上清萃取液中芯材(苯乙基间苯二酚、对羟基苯乙酮、白藜芦醇、光甘草定或虾青素)含量,计算包封率,结果如表1所示,包封率计算公式如下。
包封率(%)=[(芯材总质量-萃取芯材质量)/芯材总质量]。
表1
样品 | 包封率(%) |
实施例1 | 85.34 |
实施例2 | 88.20 |
实施例3 | 84.69 |
实施例4 | 72.47 |
实施例5 | 75.66 |
实施例6 | 70.89 |
实施例7 | 81.53 |
对比例1 | 36.61 |
对比例2 | 58.77 |
由表1可知,实施例1~7制备的环状γ-聚谷氨酸纳米微胶囊包封率均在70%以上,远远高于对比例1的36.61%和对比例2的58.77%。这说明本发明提供的环状γ-聚谷氨酸纳米微胶囊中,环状γ-聚谷氨酸成环后形成亲水及疏水空间,可以增加非水溶性物质的亲水性及稳定性,并可以与带正电的有效成分发生吸引能力,创造一个优良的包裹系统,可以有效的包裹各种芯材。特别是虾青素提取后以单酯或双酯的形式存在,具有溶脂溶性而难溶于水,通过环γ-聚谷氨酸包裹后增加其溶解性,得到透明溶液,并通过包裹来减少虾青素的氧化分解,增加稳定性,提高了包封率。
2、缓释效果测定
分别准确称取50mg实施例1~7和对比例1~2制备的纳米微胶囊样品于1.5mL离心管,然后加入1mL超纯水,混匀,然后用0.1moL/L的HCL调节pH至3.0,于1000r/min下离心30min,静置,每隔2h取样10mL加入等体积的正己烷涡旋3min,静置10min后收集正己烷测定芯材(苯乙基间苯二酚、对羟基苯乙酮、白藜芦醇、光甘草定或虾青素)含量,计算释放率,结果如表2所示。
表2
样品 | 4h释放率(%) | 8h释放率(%) |
实施例1 | 61.85 | 91.42 |
实施例2 | 55.73 | 90.54 |
实施例3 | 65.28 | 92.11 |
实施例4 | 59.37 | 87.66 |
实施例5 | 62.43 | 89.35 |
实施例6 | 60.17 | 90.27 |
实施例7 | 64.31 | 91.55 |
对比例1 | 82.65 | 88.74 |
对比例2 | 78.32 | 89.20 |
由表2可知,实施例1~7制备的环状γ-聚谷氨酸纳米微胶囊4h释放率为55~66%,8h释放率达到了87%以上,而对比例1和和对比例2的4h释放就达到了78%以上。这说明本发明提供的环状γ-聚谷氨酸纳米微胶囊中,环状γ-聚谷氨酸成环后形成亲水及疏水空间,可以增加非水溶性物质的亲水性及稳定性,并可以与带正电的有效成分发生吸引能力,创造一个优良的缓释系统,包裹后可以有效的缓释各种芯材。特别是壳聚糖可以与环γ-聚谷氨酸稳定静电吸附,进一步增强γ-聚谷氨酸包裹虾青素后的靶向性及缓释功效,避免虾青素吸收后在体内迅速代谢、消耗,增加功效持久性,同时不同组分根据特性筛选能有效结合,超出各自的原本功效。并避免了油脂及一些乳化剂的使用,通过γ-聚谷氨酸包裹也可避免添加其他抗氧化剂用于保护虾青素。产品清爽持久,具有美白、保湿、抗氧化等多种功效,可以广泛应用于食品、化妆品。
Claims (10)
1.一种环状γ-聚谷氨酸纳米微胶囊的制备方法,其特征在于,包括步骤如下:
步骤a:在避光条件下,将芯材溶于溶剂中,得到芯材溶液;将环状γ-聚谷氨酸溶于水中,得到环状γ-聚谷氨酸水溶液;将壳聚糖溶于醋酸中,得到壳聚糖溶液;
所述芯材为苯乙基间苯二酚、对羟基苯乙酮、白藜芦醇、光甘草定或虾青素中的一种或几种组合物;
所述溶剂为无水乙醇或水;
步骤b:一边搅拌,一边将芯材溶液滴加至环状γ-聚谷氨酸水溶液中,滴加完成后继续搅拌1~2h后,除去乙醇,得到浓缩溶液;
步骤c:一边搅拌,一边将浓缩溶液滴加至壳聚糖溶液中,滴加完成后继续搅拌5~10min,经过滤、冷冻、干燥后,得到环状γ-聚谷氨酸纳米微胶囊。
2.如权利要求1所述的环状γ-聚谷氨酸纳米微胶囊的制备方法,其特征在于,步骤a中,所述芯材溶液的浓度为0.02~1mg/mL。
3.如权利要求1所述的环状γ-聚谷氨酸纳米微胶囊的制备方法,其特征在于,步骤a中,所述环状γ-聚谷氨酸溶液的浓度为0.2~5mg/mL。
4.如权利要求1所述的环状γ-聚谷氨酸纳米微胶囊的制备方法,其特征在于,步骤a中,所述壳聚糖溶液的浓度为1~5mg/mL。
5.如权利要求1所述的环状γ-聚谷氨酸纳米微胶囊的制备方法,其特征在于,步骤b中,所述芯材溶液和环状γ-聚谷氨酸水溶液的体积比为(1~5):(5~15);所述搅拌速度为200~1000r/min,滴加速度为50~100mL/min。
6.如权利要求1所述的环状γ-聚谷氨酸纳米微胶囊的制备方法,其特征在于,步骤b中,通过旋转蒸发仪除去乙醇,旋蒸条件为30~40℃,真空度为1~5mbar。
7.如权利要求1所述的环状γ-聚谷氨酸纳米微胶囊的制备方法,其特征在于,步骤c中,所述搅拌速度为1500~2000r/min,滴加速度为10~50mL/min。
8.一种环状γ-聚谷氨酸纳米微胶囊,其特征在于,按照权利要求1~7任一项所述的环状γ-聚谷氨酸纳米微胶囊的制备方法制得。
9.如权利要求8所述的环状γ-聚谷氨酸纳米微胶囊,其特征在于,所述环状γ-聚谷氨酸纳米微胶囊的粒度范围是100~200nm。
10.权利要求9所述的环状γ-聚谷氨酸纳米微胶囊在制备化妆品、医美产品中的应用。
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