CN107668714A - 一种β‑聚苹果酸/壳聚糖包埋番茄红素的微胶囊及其制备方法 - Google Patents
一种β‑聚苹果酸/壳聚糖包埋番茄红素的微胶囊及其制备方法 Download PDFInfo
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Abstract
本发明公开了一种β‑聚苹果酸/壳聚糖包埋的番茄红素微胶囊及其制备方法,芯材为番茄红素乳化液,壁材为质量比为2:1的β‑聚苹果酸和壳聚糖,制备方法包括以下步骤:1)制备番茄红素乳化液;2)将所述的番茄红素乳化液缓慢地加入壳聚糖醋酸溶液;将β‑聚苹果酸水溶液滴加入番茄红素的壳聚糖醋酸溶液中,得β‑聚苹果酸/壳聚糖‑番茄红素微胶囊溶液;将β‑聚苹果酸/壳聚糖‑番茄红素微胶囊溶液通过旋转蒸发、真空冷冻干燥制得β‑聚苹果酸/壳聚糖‑番茄红素微胶囊。将β‑聚苹果酸与壳聚糖作为壁材,对番茄红素进行包埋制成微胶囊,增加了番茄红素的稳定性,使得番茄红素更广泛的应用于食品、药品、保健品、化妆品等方面。
Description
技术领域
本发明属于保健品加工技术领域,尤其是涉及一种β-聚苹果酸/壳聚糖包埋番茄红素的微胶囊及其制备方法。
背景技术
微胶囊技术,就是通过选用合适的成膜材料(壁材)将液体、固体甚至是气体(芯材)包覆形成微小颗粒的技术,芯材指被包埋的物质;壁材指用于成型的物质。芯材为油溶性化合物、水溶性化合物或两者的混合物,芯材分为固体、液体和气体。芯材与壁材的溶解性能必须不一样,即水溶性芯材用油溶性壁材,而油溶性芯材用水溶性壁材。微胶囊技术是一项比较新颖,用途广泛,发展迅速的新技术。微胶囊处理后,芯材不与外界环境因素直接接触,使芯材得到有效保护,同时还能屏蔽味道和气味,降低对健康的危害、减少毒副作用等。
聚苹果酸(poly(β-)malic acid),简称PMLA,是一种以L-苹果酸为唯一单体,通过-COOH和-OH基团聚合而成的阴离子聚合物。β-聚苹果酸的侧链上含有若干个亲水性极强的羧基基团,使得PMLA具有较强的溶解性,能够与水混溶和吸收水分。其次,羧基还是一种弱酸性官能团使得β-聚苹果酸具有一定的弱酸性。PMLA具有高度的水溶性、吸水性、可吸收性、生物相容性、生物可降解性、可化学衍生性、无毒性及无免疫原性等。可在水溶液中自发或由酶促降解为小分子苹果酸,可被人体吸收并无任何毒副作用,β-聚苹果酸本身带负电,具有阴离子的性质,可与阳离子结合。将PMLA应用到药物中可提高药物药效并降低毒副作用。
壳聚糖是由自然界中广泛存在的几丁质经过脱乙酰作用得到的,其活性基团为脱乙酰基后的氨基,由于其分子结构中存在一定量的氨基,使壳聚糖具有弱碱性,同时壳聚糖是目前唯一的天然的阳离子聚合物。这种天然高分子的生物官能性和生物相容性、安全性、微生物降解性等优良性能被各行各业广泛关注,在医药、食品、化工、化妆品、水处理、生化和生物医学工程等诸多领域的应用研究取得了重大进展。
番茄红素(Lycopene,又称ψ—胡萝卜素)属于异戊二烯类化合物,是类胡萝卜素的一种,广泛存在于番茄、西瓜、葡萄、胡萝卜等植物中。它是目前在自然界的植物中被发现的最强抗氧化剂之一。科学证明,人体内的单线态氧和氧自由基是侵害人体自身免疫系统的罪魁祸首。番茄红素清除自由基的功效远胜于其他类胡萝卜素和维生素E,其淬灭单线态氧速率常数是维生素E的100倍,是β-胡萝卜素的两倍多。它可以有效的防治因衰老,免疫力下降引起的各种疾病。
然而,番茄红素是脂溶性色素,可溶于其他脂类和非极性溶剂中,不溶于水,限制了其在食品、药品中的应用。同时,番茄红素分子中有11个共轭双键和2个非共轭双键,使得番茄红素的稳定性比较差,易被氧化分解,其在一定条件下还可能发生顺反异构化。番茄红素含有大量不饱和结构,在对番茄红素的提取分离、加工处理和储存过程中,由于热、光、酸、碱、表面活性剂、氧化剂和还原剂等均可以促进这些变化,导致其生理活性降低,更限制了番茄红素的广泛应用。
发明内容
为解决上述存在的技术问题,本发明的目的是提供一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊及其制备方法。将β-聚苹果酸与壳聚糖作为壁材,对番茄红素进行包埋制成微胶囊,增加了番茄红素的稳定性,使得番茄红素更广泛的应用于食品、药品、保健品、化妆品等方面。
为实现上述目的,本发明的技术方案如下:
一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊,芯材为番茄红素乳化液,壁材为质量比为2:1的β-聚苹果酸和壳聚糖。
一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊的制备方法,包括以下步骤
1)制备番茄红素乳化液;
2)将所述番茄红素乳化液缓慢地加入到0.5~1mg/mL的壳聚糖醋酸溶液中,番茄红素乳化液:壳聚糖醋酸溶液的比例为1:1~3(v/v),控制搅拌速度为400~600r/min,搅拌30s后得到番茄红素的壳聚糖醋酸溶液;
3)将1~2mg/mL的β-聚苹果酸水溶液滴加入番茄红素的壳聚糖醋酸溶液中,滴加量按体积比为β-聚苹果酸溶液和壳聚糖醋酸溶液:番茄红素乳化液为15:3~5,控制滴加速率为5~10ml/h,控制搅拌速度为400~600r/min,得β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液;
4)将β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液通过旋转蒸发的方法,微胶囊溶液温度为60℃,蒸发出混合液中多余的有机溶剂,再进行真空冷冻干燥制得β-聚苹果酸/壳聚糖-番茄红素微胶囊。
本发明中番茄红素乳化液的制备方法为:将番茄红素粉末溶于乙酸乙酯中,在60℃下进行搅拌溶解得到油相,并加入吐温80作为乳化剂使之分散均匀,制得番茄红素乳化液,所述的番茄红素粉末和乙酸乙酯比例为1:10(m/v),乙酸乙酯与吐温的比例为100:1(v/v)。
更进一步的,β-聚苹果酸水溶液为将β-聚苹果酸溶于去离子水中,配成1~2mg/mL的β-聚苹果酸水溶液。
更进一步的,壳聚糖醋酸溶液为将壳聚糖溶于1%的冰乙酸溶液中,配成0.5~1mg/mL的壳聚糖醋酸溶液。
更进一步的,在步骤3)中用蠕动泵控制β-聚苹果酸水溶液滴加速率为6-8ml/h,用磁力搅拌器控制搅拌速度为450~500r/min,得β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液。
本发明中的β-聚苹果酸的制备方法为:将出芽短梗霉A. pullulans CGMCC3337菌株活化后按照10%(v/v)的接种量接入到种子培养基中,于25 ℃,200 r/min,培养40个小时至对数生长期;然后将种子培养基接入到发酵培养基中,接种量为10%,发酵罐为5L,装液量为60%,发酵条件:恒定发酵温度25 ℃,通风比1:1.2,转速为450rpm,发酵时间144 h;将发酵液用800目滤布预涂硅藻土,硅藻土加入量为1%(m/v),于0.15~0.2Mpa下进行板框过滤除菌体,将除菌体后的发酵液加热到60℃后加入1.5%活性炭脱色60min,然后将脱色后的发酵液用截留分子量10KDa的聚醚砜超滤膜于40℃,0.2Mpa进行超滤除去多糖及大分子杂质,用300Da的复合聚酰胺纳滤膜于40℃,0.8Mpa进行纳滤除去寡糖、盐离子等小分子杂质,最后通过旋蒸浓缩、冷冻干燥即得β-聚苹果酸。
本发明技术方案的机理:
β-聚苹果酸本身带负电,具有阴离子的性质,可与阳离子结合;壳聚糖是由自然界中广泛存在的几丁质经过脱乙酰作用得到的,其活性基团为脱乙酰基后的氨基,由于其分子结构中存在一定量的氨基,使壳聚糖具有弱碱性,同时壳聚糖是目前唯一的天然的阳离子聚合物。β-聚苹果酸可与带正电壳聚糖发生絮凝形成微胶囊。
本发明的有益效果是:
1、本发明的β-聚苹果酸/壳聚糖-番茄红素微胶囊是利用β-聚苹果酸和壳聚糖为壁材原料制备而成的纳米级微胶囊,而β-聚苹果酸和壳聚糖都是未经过修饰的天然产物,具有很好的生物相容性和生物可降解性,因此以此为壁材制备微胶囊对人体和环境无毒无害。
2、本发明采用对人体无毒、遇水易释放芯材的物质作为壁材,并采用适宜的微胶囊化方法将番茄红素包覆在微小且封闭的膜内,使之转化为易处理的粉末固体,保护敏感的番茄红素,防止其被氧化分解,生理活性降低。
3、本发明采用的β-聚苹果酸一类新型的生物高分子材料,具有特殊的立体结构,主链上带有酯基,在水环境条件下可发生自发或酶促降解。采用β-聚苹果酸和壳聚糖发生复凝聚制作的壁材可以实现芯材的缓慢释放,延长功能因子的作用时间,使番茄红素的稳定性得到显著提高,而且还能改善番茄红素微胶囊的水溶性,进而扩大应用领域。
4、采用低温旋蒸和冷冻干燥的精制方法,进一步保护了敏感的番茄红素,防止氧化、降解。
5、本发明的技术方案包埋的番茄红素微胶囊,质量稳定,微胶囊中包埋的番茄红素有效物质含量在32%左右,经光照八周后,番茄红素有效物质仍能达到20%以上。有效减缓了番茄红素随光照时间的降解速度。
具体实施方式
实施例1
(1)、β-聚苹果酸的制备
将出芽短梗霉A. pullulans CGMCC3337菌株活化后按照10%(v/v)的接种量接入到种子培养基中,于25 ℃,200 r/min,培养40个小时至对数生长期。然后将种子培养基接入到发酵培养基中,接种量为10%,发酵罐为5L,装液量为60%,发酵条件:恒定发酵温度25 ℃,通风比1:1.2,转速为450rpm,发酵时间144 h。
将发酵液用800目滤布预涂硅藻土,硅藻土加入量为1%(m/v),于0.15~0.2Mpa下进行板框过滤除菌体,将除菌体后的发酵液加热到60℃后加入1.5%活性炭脱色60min,然后将脱色后的发酵液用截留分子量10KDa的聚醚砜超滤膜于40℃,0.2Mpa进行超滤除去多糖及大分子杂质,用300Da的复合聚酰胺纳滤膜于40℃,0.8Mpa进行纳滤除去寡糖、盐离子等小分子杂质,最后通过旋蒸浓缩、冷冻干燥即得β-聚苹果酸。
(2)溶液的配制
将β-聚苹果酸溶于去离子水中,配成1mg/mL的β-聚苹果酸水溶液,pH为自然值;
将壳聚糖溶于1%的冰乙酸溶液中,配成1mg/mL的壳聚糖醋酸溶液;
将5g番茄红素粉末溶于50mL乙酸乙酯中,在60℃下进行搅拌溶解得到油相,并加入0.5mL吐温80作为乳化剂使之分散均匀。
(3)将步骤(2)中所得的番茄红素乳化液3mL缓慢加入5mL壳聚糖醋酸溶液中,控制搅拌速度为500r/min,搅拌30s后得到番茄红素的壳聚糖醋酸溶液。β-聚苹果酸和壳聚糖按质量比为2:1,壁材总量为15mL,番茄红素乳液的添加量为3mL。
通过边滴加边反应的方式将10mLβ-聚苹果酸水溶液加入到上述所得番茄红素的壳聚糖醋酸溶液中,用蠕动泵控制滴加速率在10ml/h,用磁力搅拌器控制搅拌速度为500r/min搅拌1h,得β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液。
(4)考虑到番茄红素的热敏性,将步骤(3)所得的β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液通过旋转蒸发的方法,微胶囊溶液温度为60℃,蒸发出混合液中多余的有机溶剂,再进行真空冷冻干燥制得β-聚苹果酸/壳聚糖-番茄红素微胶囊。
本实施例番茄红素微胶囊稳定性试验及检测结果:
将番茄红素原料、制备好的番茄红素微胶囊分别装于透明的样品瓶中密封保存,在室温(20±5℃)下采用自然光照射,每隔一段时间测定样品中番茄红素的含量,与贮存前样品中的番茄红素含量相比得番茄红素保留率。表1为不同样品中番茄红素含量的测定结果。
表1
样品 时间 | 起始 | 1周 | 2周 | 4周 | 8周 |
番茄红素原料 | 98% | 30.4% | 7.8% | 2.4% | 0 |
本实施例中番茄红素微胶囊 | 32.1% | 29.5% | 26.8% | 23.4% | 20.6% |
实施例2
(1)、β-聚苹果酸的制备
将出芽短梗霉A. pullulans CGMCC3337菌株活化后按照10%(v/v)的接种量接入到种子培养基中,于25 ℃,200 r/min,培养40个小时至对数生长期。然后将种子培养基接入到发酵培养基中,接种量为10%,发酵罐为5L,装液量为60%,发酵条件:恒定发酵温度25 ℃,通风比1:1.2,转速为450rpm,发酵时间144 h。
将发酵液用800目滤布预涂硅藻土,硅藻土加入量为1%(m/v),于0.15~0.2Mpa下进行板框过滤除菌体,将除菌体后的发酵液加热到60℃后加入1.5%活性炭脱色60min,然后将脱色后的发酵液用截留分子量10KDa的聚醚砜超滤膜于40℃,0.2Mpa进行超滤除去多糖及大分子杂质,用300Da的复合聚酰胺纳滤膜于40℃,0.8Mpa进行纳滤除去寡糖、盐离子等小分子杂质,最后通过旋蒸浓缩、冷冻干燥即得β-聚苹果酸。
(2)溶液的配制
将β-聚苹果酸溶于去离子水中,配成2mg/mL的β-聚苹果酸水溶液,pH为自然值;
将壳聚糖溶于1%的冰乙酸溶液中,配成1mg/mL的壳聚糖醋酸溶液;
将5g番茄红素粉末溶于50mL乙酸乙酯中,在60℃下进行搅拌溶解得到油相,并加入1mL吐温80作为乳化剂使之分散均匀。
(3)将步骤(2)中所得的番茄红素乳液4mL缓慢加入7.5mL壳聚糖醋酸溶液中,控制搅拌速度为600r/min,搅拌30s后得到番茄红素的壳聚糖醋酸溶液;
将β-聚苹果酸和壳聚糖按质量比为2:1进行反应,壁材总量为15mL,番茄红素乳液的添加量为4mL。
通过边滴加边反应的方式将7.5mL聚苹果酸水溶液加入到上述所得番茄红素的壳聚糖醋酸溶液中,用蠕动泵控制滴加速率在7.5ml/h,用磁力搅拌器控制搅拌速度为600r/min,得β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液。
(4)考虑到番茄红素的热敏性,将步骤(3)所得的β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液通过旋转蒸发的方法,微胶囊溶液温度为60℃,蒸发出混合液中多余的有机溶剂,再进行真空冷冻干燥制得β-聚苹果酸/壳聚糖-番茄红素微胶囊。
本实施例中番茄红素原料及番茄红素微胶囊中番茄红素的稳定性检测结果见表2。
表2
样品 时间 | 起始 | 1周 | 2周 | 4周 | 8周 |
番茄红素原料 | 98% | 30.4% | 7.8% | 2.4% | 0 |
本实施例中番茄红素微胶囊 | 32.1% | 28.5% | 26.8% | 24.4% | 21.6% |
实施例3
(1)、β-聚苹果酸的制备
将出芽短梗霉A. pullulans CGMCC3337菌株活化后按照10%(v/v)的接种量接入到种子培养基中,于25 ℃,200 r/min,培养40个小时至对数生长期。然后将种子培养基接入到发酵培养基中,接种量为10%,发酵罐为5L,装液量为60%,发酵条件:恒定发酵温度25 ℃,通风比1:1.2,转速为450rpm,发酵时间144 h。
将发酵液用800目滤布预涂硅藻土,硅藻土加入量为1%(m/v),于0.15~0.2Mpa下进行板框过滤除菌体,将除菌体后的发酵液加热到60℃后加入1.5%活性炭脱色60min,然后将脱色后的发酵液用截留分子量10KDa的聚醚砜超滤膜于40℃,0.2Mpa进行超滤除去多糖及大分子杂质,用300Da的复合聚酰胺纳滤膜于40℃,0.8Mpa进行纳滤除去寡糖、盐离子等小分子杂质,最后通过旋蒸浓缩、冷冻干燥即得β-聚苹果酸。
(2)溶液的配制
将β-聚苹果酸溶于去离子水中,配成1.5mg/mL的β-聚苹果酸水溶液,pH为自然值;
将壳聚糖溶于1%的冰乙酸溶液中,配成0.5mg/mL的壳聚糖醋酸溶液;
将5g番茄红素粉末溶于50mL乙酸乙酯中,在60℃下进行搅拌溶解得到油相,并加入1.5mL吐温80作为乳化剂使之分散均匀。
(3)将步骤(2)中所得的番茄红素乳液5mL缓慢加入9mL壳聚糖醋酸溶液中,控制搅拌速度为400r/min,搅拌30s后得到番茄红素的壳聚糖醋酸溶液;
将β-聚苹果酸和壳聚糖按质量比为2:1进行反应,壁材总量为15mL,番茄红素乳液的添加量为5mL。
通过边滴加边反应的方式将6mLβ-聚苹果酸水溶液加入到上述所得番茄红素的壳聚糖醋酸溶液中,用蠕动泵控制滴加速率在6ml/h,用磁力搅拌器控制搅拌速度为400r/min搅拌1h,得β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液。
(4)考虑到番茄红素的热敏性,将步骤(3)所得的β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液通过旋转蒸发的方法,微胶囊溶液温度为60℃,蒸发出混合液中多余的有机溶剂,再进行真空冷冻干燥制得β-聚苹果酸/壳聚糖-番茄红素微胶囊。
本实施例中番茄红素原料及番茄红素微胶囊中番茄红素的稳定性检测结果见表3。
表3
样品 时间 | 起始 | 1周 | 2周 | 4周 | 8周 |
番茄红素原料 | 98% | 30.4% | 7.8% | 2.4% | 0 |
本实施例中番茄红素微胶囊 | 32.1% | 29.5% | 28.2% | 26.4% | 21.5% |
以上实施例仅用以说明本发明的技术方案而非限制,尽管参照较佳实施例对本发明进行了详细说明,本领域的普通技术人员对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的宗旨和范围,其均应涵盖在本发明的权利要求范围当中。
Claims (7)
1.一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊,其特征在于,芯材为番茄红素乳化液,壁材为质量比为2:1的β-聚苹果酸和壳聚糖。
2.一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊的制备方法,其特征在于,包括以下步骤
1)制备番茄红素乳化液;
2)将所述番茄红素乳化液缓慢地加入到0.5~1mg/mL的壳聚糖醋酸溶液中,番茄红素乳化液:壳聚糖醋酸溶液的比例为1:1~3(v/v),控制搅拌速度为400~600r/min,搅拌30s后得到番茄红素的壳聚糖醋酸溶液;
3)将1~2mg/mL的β-聚苹果酸水溶液滴加入番茄红素的壳聚糖醋酸溶液中,滴加量按体积比为β-聚苹果酸溶液和壳聚糖醋酸溶液:番茄红素乳化液为15:3~5,控制滴加速率为5~10ml/h,控制搅拌速度为400~600r/min,得β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液;
4)将β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液通过旋转蒸发的方法,微胶囊溶液温度为60℃,蒸发出混合液中多余的有机溶剂,再进行真空冷冻干燥制得β-聚苹果酸/壳聚糖-番茄红素微胶囊。
3.根据权利要求2所述的一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊的制备方法,其特征在于,所述的番茄红素乳化液的制备方法为:将番茄红素粉末溶于乙酸乙酯中,在60℃下进行搅拌溶解得到油相,并加入吐温80作为乳化剂使之分散均匀,制得番茄红素乳化液,所述的番茄红素粉末和乙酸乙酯比例为1:10(m/v),乙酸乙酯与吐温的比例为100:1(v/v)。
4.根据权利要求2所述的一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊的制备方法,其特征在于,所述的β-聚苹果酸水溶液为将β-聚苹果酸溶于去离子水中,配成1~2mg/mL的β-聚苹果酸水溶液。
5.根据权利要求2所述的一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊的制备方法,其特征在于,所述的壳聚糖醋酸溶液为将壳聚糖溶于1%的冰乙酸溶液中,配成0.5~1mg/mL的壳聚糖醋酸溶液。
6.根据权利要求2所述的一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊的制备方法,其特征在于,步骤3)中用蠕动泵控制β-聚苹果酸水溶液滴加速率为6-8ml/h,用磁力搅拌器控制搅拌速度为450~500r/min搅拌1h,得β-聚苹果酸/壳聚糖-番茄红素微胶囊溶液。
7.根据权利要求2至6中的任一项权利要求中所述的一种β-聚苹果酸/壳聚糖包埋的番茄红素微胶囊的制备方法,其特征在于,所述的β-聚苹果酸的制备方法为:将出芽短梗霉A.pullulans CGMCC3337菌株活化后按照10%(v/v)的接种量接入到种子培养基中,于25 ℃,200 r/min,培养40个小时至对数生长期;然后将种子培养基接入到发酵培养基中,接种量为10%,发酵罐为5L,装液量为60%,发酵条件:恒定发酵温度25 ℃,通风比1:1.2,转速为450rpm,发酵时间144 h;将发酵液用800目滤布预涂硅藻土,硅藻土加入量为1%(m/v),于0.15~0.2Mpa下进行板框过滤除菌体,将除菌体后的发酵液加热到60℃后加入1.5%活性炭脱色60min,然后将脱色后的发酵液用截留分子量10KDa的聚醚砜超滤膜于40℃,0.2Mpa进行超滤除去多糖及大分子杂质,用300Da的复合聚酰胺纳滤膜于40℃,0.8Mpa进行纳滤除去寡糖、盐离子等小分子杂质,最后通过旋蒸浓缩、冷冻干燥即得β-聚苹果酸。
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