CN108720022A - 一种花色苷纳米脂质体的制备方法和应用 - Google Patents
一种花色苷纳米脂质体的制备方法和应用 Download PDFInfo
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Abstract
本发明公开了一种花色苷纳米脂质体的制备方法,包括以下步骤:将大豆卵磷脂、胆固醇、吐温‑80和花色苷溶解于无水乙醇中,再加入缓冲溶液,搅拌反应后蒸发去除乙醇,形成脂质体分散液,脂质体分散液经过超声处理,得到所述花色苷纳米脂质体。本发明花色苷纳米脂质体的制备方法工艺简单,容易实施,能够有效稳定花色苷,抑制花色苷的分解和氧化,而且相比现有技术,其包封率更高,粒径更小。
Description
技术领域
本发明涉及花色苷的应用领域,具体地说涉及一种花色苷纳米脂质体的制备方法和应用。
背景技术
花色苷是一种水溶性天然食用色素,广泛存在于自然界,是花青素和糖类以不同方式结合形成的黄酮多酚类化合物。花色苷常作为天然食品着色剂,安全无毒,且具有多种生理功效,例如抗氧化性,清除自由基功能,抗肿瘤,抗炎,抗菌,预防心脑血管疾病,降血脂,改善视力等,被誉为继水、蛋白质、脂肪、碳水化合物、维生素、矿物质之后的第七大必需营养素,继而容易被广大人群所接受。尽管花色苷在食品行业应用潜力巨大,但是其本身的稳定性很容易受到外界环境的影响,对pH、光照、温度等外界条件都较敏感,所以大大限制了其在各个领域的广泛运用。
近年来,随着生物科技的不断进步,脂质体作为载体包埋生物活性物质研究得到关注,脂质体正日益成为一种新型理想载体。脂质体是由双亲性(亲水的极性头部和疏水的非极性尾部)脂质双分子层包裹某些活性物质后自组装形成的具有双层膜结构的微型泡囊。脂质体作为载体具有双亲性、生物相容性、低毒性、保护性、靶向性和缓释性等特点。脂质体研究初期多作为药物的载体,近年来在食品领域逐渐得到应用,起初脂质体大都用来包埋酶来促进奶酪的熟化,如今脂质体在营养素的包埋和稳定方面的研究逐渐成为关注的热点,但对营养素花色苷的研究仍较少。
然而,现有技术在制备花色苷纳米脂质体时,包封率较低,粒径也相对较大,都高于 100nm。例如,申请号为201210177952.1的中国专利申请公开的花色苷提取物脂质体及其制备方法,采用逆相蒸发法制备花色苷提取物脂质体,其包封率不到40%,粒径超过了10μ m,申请号为201610054330.8的中国专利申请公开的杨梅花色苷纳米脂质体的制备方法,采用逆向蒸发法制备杨梅花色苷脂质体,其包封率不到60%,粒径为100-1000nm。以上两个专利并未涉及到花色苷纳米脂质体的应用方面,而本发明使用了乙醇注入法制备花色苷纳米脂质体,并进一步对其在食品领域在的应用进行了阐述。
发明内容
本发明所要解决的技术问题是提供一种花色苷纳米脂质体的制备方法,该方法包封率更高,产品粒径更小。
为了解决上述技术问题,本发明采用如下技术方案:一种花色苷纳米脂质体的制备方法,包括以下步骤:
将大豆卵磷脂、胆固醇、吐温-80和花色苷溶解于无水乙醇中,再加入缓冲溶液,搅拌反应后蒸发去除乙醇,形成脂质体分散液,脂质体分散液经过超声处理,得到所述花色苷纳米脂质体。
进一步地,所述缓冲溶液为pH=4.0、浓度为0.05mol/L的乙酸-乙酸钠缓冲溶液。在实施本发明的过程中,发明人发现,采用这种缓冲溶液,能够保证获得更高的包封率和更小的粒径。
进一步地,大豆卵磷脂、胆固醇、吐温-80、花色苷、无水乙醇与缓冲溶液的质量体积比为0.8~1.2(g):0.15~0.20(g):0.20~0.30(g):0.06~0.20(g):80~120(mL): 80~120(mL)。在实施本发明的过程中,发明人发现,采用以上配比,能够保证获得更高的包封率和更小的粒径。
进一步地,超声处理的功率密度为3.0~5.0W/cm3,时间为3~7min。在实施本发明的过程中,发明人发现,采用以上条件,能够保证获得更高的包封率和更小的粒径。
进一步地,蒸发去除乙醇在40℃条件下进行。在实施本发明的过程中,发明人发现,采用这个条件,能够保证获得更高的包封率和更小的粒径。
进一步地,搅拌反应的时间为0.5h。在实施本发明的过程中,发明人发现,采用这个条件,能够保证获得更高的包封率和更小的粒径。
本发明还提供一种巧克力,按重量份,包括以下原料:可可液块100~160、可可脂100~150、白糖100~150、奶粉100~150、按上述花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体15~25。
本发明还提供一种奶片,按重量份,包括以下原料:全脂奶粉100~150、柠檬酸2~6、麦芽糊精20~30、硬脂酸镁0.5~1.0、按上述花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体10~20。
本发明还提供一种曲奇,按重量份,包括以下原料:低筋面粉100~150、鸡蛋15~25、黄油40~60、奶油10~20、糖粉20~40、细砂糖10~20、按上述花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体10~20。
本发明还提供一种牛奶,按重量份,包括以下原料:生牛乳100~150、白糖10~20、按上述花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体10~20。
本发明还提供一种奶茶,按重量份,包括以下原料:全脂奶粉40~60份、绿茶粉10~20、白糖5~8、水40~80、按上述花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体5~10。
本发明的有益效果体现在:
本发明花色苷纳米脂质体的制备方法简单,容易实施,能够有效稳定花色苷,抑制花色苷的分解和氧化,而且相比现有技术,其包封率更高,粒径更小,脂质体包封率和粒径是评价脂质体质量和应用价值的重要指标。本发明制得的花色苷纳米脂质体包封率高,说明有更多的花色苷得到了脂质双层保护,进而就能更显著的减缓花色苷的降解速率。另外,花色苷纳米脂质体的粒径大小直接影响其释放、生物利用率以及应用。有文献报道在500nm或以下的纳米颗粒更适合于食品应用,而且粒径越小,有更高的稳定性和生物利用度。上述制备的花色苷纳米脂质体有较好的稳定性,因此可用于巧克力、奶片、曲奇、牛奶、奶茶等食品中,不仅可赋予以上食品良好的色泽特性,还提高了这些食品的营养功能特性。
附图说明
图1是实施例1制得的花色苷纳米脂质体与对照组花色苷溶液的温度稳定性比较图。
图2是温度对实施例1制得的花色苷纳米脂质体的影响图。
图3是实施例1制得的花色苷纳米脂质体与对照组花色苷溶液的光稳定性比较图。
图4是光照对实施例1制得的花色苷纳米脂质体的影响图。
图5是实施例1制得的花色苷纳米脂质体与对照组花色苷溶液的pH稳定性比较图。
图6是pH对实施例1制得的花色苷纳米脂质体的影响图。
具体实施方式
下面结合实施例对本发明作进一步描述:
以下实施例所使用的各种原料,如未作特别说明,均为本领域公知的市售产品。
实施例1
花色苷纳米脂质体的制备
将1.08g大豆卵磷脂、0.18g胆固醇、0.24g吐温-80、0.15g蓝莓花色苷溶解在100mL无水乙醇中,再加入100mL的pH=4.0、浓度为0.05mol/L的乙酸-乙酸钠缓冲溶液,在磁力搅拌器上反应0.5h后转移到旋转蒸发器中于40℃下蒸发去除乙醇,形成脂质体分散液,脂质体分散液在4.4W/cm3功率密度下超声处理5min,得到所述花色苷纳米脂质体。
本实施制得的花色苷纳米脂质体包封率为91.13%,粒径为53.80nm。
实施例2
花色苷纳米脂质体的制备
将0.8g大豆卵磷脂、0.15g胆固醇、0.20g吐温-80、0.06g蓝莓花色苷溶解在80mL无水乙醇中,再加入80mL的pH=4.0、浓度为0.05mol/L的乙酸-乙酸钠缓冲溶液,在磁力搅拌器上反应0.5h后转移到旋转蒸发器中于40℃下蒸发去除乙醇,形成脂质体分散液,脂质体分散液在5.0W/cm3功率密度下超声处理3min,得到所述花色苷纳米脂质体。
本实施制得的花色苷脂质体包封率为89.76%,粒径为67.23nm。
实施例3
花色苷纳米脂质体的制备
将1.2g大豆卵磷脂、0.20g胆固醇、0.30g吐温-80、0.20g蓝莓花色苷溶解在120mL无水乙醇中,再加入120mL的pH=4.0、浓度为0.05mol/L的乙酸-乙酸钠缓冲溶液,在磁力搅拌器上反应0.5h后转移到旋转蒸发器中于40℃下蒸发去除乙醇,形成脂质体分散液,脂质体分散液在3.0W/cm3功率密度下超声处理7min,得到所述花色苷纳米脂质体。
本实施制得的花色苷脂质体包封率为88.95%,粒径为60.27nm。
实施例4
巧克力的制备
按重量份,包括以下原料:可可液块100~160、可可脂100~150、白糖100~150、奶粉100~150、实施例1制得的花色苷纳米脂质体15~25。
制备步骤:
(1)基料混合:将可可液块、可可脂、白糖、奶粉混合,然后在加热条件下充分搅拌使配料完全融化,随后加入花色苷纳米脂质体,混合均匀,得到基料;
(2)冷却成型:将基料趁热倒入模具中,并在室温下进行冷却。
加入花色苷纳米脂质体制备而成的含花色苷的巧克力除具有巧克力的营养价值外,还富含丰富且稳定的花色苷成分,进一步提高了含花色苷巧克力的营养及功能特性。
实施例5
奶片的制备
按重量份,包括以下原料:全脂奶粉100~150、柠檬酸2~6、麦芽糊精20~30、硬脂酸镁0.5~1.0、实施例2制得的花色苷纳米脂质体10~20。
制备步骤:
(1)基料的制备:将全脂奶粉、柠檬酸、麦芽糊精、硬脂酸镁、花色苷纳米脂质体混合,搅拌均匀,得到基料;
(2)造粒与干燥:将基料投入摇摆式颗粒机中进行造粒,得到的湿颗粒置于30℃鼓风干燥箱中干燥,20min翻动一次,直至颗粒中的水分含量降至3%;
(3)压片:将干燥后的颗粒送入压片机压制成片。
加入花色苷纳米脂质体制备而成的含花色苷的奶片除具有奶片的营养价值外,还富含丰富且稳定的花色苷成分,进一步提高了含花色苷奶片的营养及功能特性。另外,花色苷纳米脂质体的加入改善了奶片的外观色泽,且奶片颜色呈粉红色。
实施例6
曲奇的制备
按重量份,包括以下原料:低筋面粉100~150、鸡蛋15~25、黄油40~60、奶油10~20、糖粉20~40、细砂糖10~20、实施例3制得的花色苷纳米脂质体10~20。
制备步骤:
(1)原料制备:取黄油在室温下放至软化,然后搅打至顺滑状态,取细砂糖、糖粉加入到软化后的黄油中,继续搅拌至黄油颜色变浅且顺滑的状态,分三次加入打散的蛋液及奶油,然后打发至融合状态,最后加入低筋面粉,搅拌均匀,再加入花色苷纳米脂质体,拌匀成面团;
(2)曲奇雏形制备:将拌好的面团装入裱花袋中,整齐的挤在铺有油纸的烤盘上;
(3)烘烤:烤箱预热到150~180℃,烘烤10~15分钟,取出冷却制得成品。
加入花色苷纳米脂质体制备而成的含花色苷的曲奇除具有曲奇的营养价值外,还富含丰富且稳定的花色苷成分,进一步提高了含花色苷曲奇的营养及功能特性。另外,花色苷纳米脂质体的加入改善了曲奇的外观色泽。
实施例7
牛奶的制备
按重量份,包括以下原料:生牛乳100~150、白糖10~20、实施例1制得的花色苷纳米脂质体10~20。
制备步骤:
(1)牛奶的制备:将生牛乳、白糖、花色苷纳米脂质体混合均匀,得到奶液;
(2)灭菌及灌装:将混合均匀的奶液灌装到瓶中,进行巴氏杀菌,杀菌后及时冷却。
加入花色苷纳米脂质体制备而成的含花色苷的牛奶除具有牛奶的营养价值外,还富含丰富且稳定的花色苷成分,进一步提高了含花色苷牛奶的营养及功能特性。另外,花色苷纳米脂质体的加入改善了牛奶的外观色泽,且牛奶颜色呈粉红色。
实施例8
本发明还提供一种奶茶,按重量份,包括以下原料:全脂奶粉40~60份、绿茶粉10~ 20、白糖5~8、水40~80、实施例1制得的花色苷纳米脂质体5~10。
制备步骤:
(1)奶茶制备:将全脂奶粉、绿茶粉、白糖在热水中充分溶解,加入花色苷纳米脂质体,搅拌均匀,得到奶茶;
(2)灭菌及灌装:将混合均匀的奶茶灌装到瓶中,进行巴氏杀菌,杀菌后及时冷却。
加入花色苷纳米脂质体的制备而成的含花色苷的奶茶除具有牛奶的营养价值外,还富含丰富且稳定的花色苷成分,进一步提高了含花色苷奶茶的营养及功能特性。另外,花色苷纳米脂质体的加入改善了奶茶的外观色泽。
实施例9
花色苷纳米脂质体的稳定性试验
9.1温度稳定性实验
将实施例1制得的花色苷纳米脂质体与对照组花色苷溶液(相同浓度的蓝莓花色苷的水溶液,下同)做对比,比较两者在4℃/25℃下28天内花色苷的稳定性。并且对花色苷纳米脂质体的粒径和包封率变化做出判断。
9.2光稳定性实验
将实施例1制得的花色苷纳米脂质体与对照组花色苷溶液做对比,比较两者在避光/ 光照下28天内花色苷的稳定性。并且对花色苷纳米脂质体的粒径和包封率变化做出判断。
9.3pH稳定性实验
将实施例1制得的花色苷纳米脂质体与对照组花色苷溶液做对比,比较两者在 pH=3.0/5.0/7.0下5天内花色苷的稳定性。并且对花色苷纳米脂质体的粒径和包封率变化做出判断。
从图1至图6可以看出,本发明制得的花色苷纳米脂质体相对对照组,其花色苷在4℃或25℃下、避光或光照、pH3.0~7.0下保留率较高,花色苷的稳定性得到显著提高。另外,温度、光照、pH对本发明制得的花色苷纳米脂质体的粒径和包封率的影响并不是很大,包封率几乎没有太大变化,粒径变化也在1~30nm以内。综上,本发明制得的花色苷纳米脂质体是较稳定的体系,可以广泛应用在食品体系中。
应当理解本文所述的例子和实施方式仅为了说明,并不用于限制本发明,本领域技术人员可根据它做出各种修改或变化,凡在本发明的精神和原则之内,所作的任何修改、等同替换、改进等,均应包含在本发明的保护范围之内。
Claims (9)
1.一种花色苷纳米脂质体的制备方法,其特征在于,包括以下步骤:
将大豆卵磷脂、胆固醇、吐温-80和花色苷溶解于无水乙醇中,再加入缓冲溶液,搅拌反应后蒸发去除乙醇,形成脂质体分散液,脂质体分散液经过超声处理,得到所述花色苷纳米脂质体。
2.如权利要求1所述的花色苷纳米脂质体的制备方法,其特征在于:所述缓冲溶液为pH=4.0、浓度为0.05mol/L的乙酸-乙酸钠缓冲溶液。
3.如权利要求2所述的花色苷纳米脂质体的制备方法,其特征在于:大豆卵磷脂、胆固醇、吐温-80、花色苷、无水乙醇与缓冲溶液的质量体积比为0.8~1.2(g):0.15~0.20(g):0.20~0.30(g):0.06~0.20(g):80~120(mL):80~120(mL)。
4.如权利要求1或2或3所述的花色苷纳米脂质体的制备方法,其特征在于:超声处理的功率密度为3.0~5.0W/cm3,时间为3~7min。
5.一种巧克力,其特征在于,按重量份,包括以下原料:可可液块100~160、可可脂100~150、白糖100~150、奶粉100~150、按如权利要求1至4中任一项所述的花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体15~25。
6.一种奶片,其特征在于,按重量份,包括以下原料:全脂奶粉100~150、柠檬酸2~6、麦芽糊精20~30、硬脂酸镁0.5~1.0、按如权利要求1至4中任一项所述的花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体10~20。
7.一种曲奇,其特征在于,按重量份,包括以下原料:低筋面粉100~150、鸡蛋15~25、黄油40~60、奶油10~20、糖粉20~40、细砂糖10~20、按如权利要求1至4中任一项所述的花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体10~20。
8.一种牛奶,其特征在于,按重量份,包括以下原料:生牛乳100~150、白糖10~20、按如权利要求1至4中任一项所述的花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体10~20。
9.一种奶茶,其特征在于,按重量份,包括以下原料:全脂奶粉40~60份、绿茶粉10~20、白糖5~8、水40~80、按如权利要求1至4中任一项所述的花色苷纳米脂质体的制备方法制备得到的花色苷纳米脂质体5~10。
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