CN111012920A - 玉屏风多糖脂质体及其制备方法 - Google Patents
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Abstract
本发明公开了一种玉屏风多糖脂质体的制备方法，包括以下步骤：1)将大豆卵磷脂和胆固醇溶于氯仿，减压旋转蒸发，使溶液在容器内壁形成一层淡黄色脂质薄膜，随后加入***将薄膜溶解，加入玉屏风多糖和PH值7.4的磷酸盐缓冲液，进行超声处理充分混匀，随后减压抽吸至***挥发干净，即得玉屏风多糖脂质体基体；2)将玉屏风多糖脂质体基体按照1:1的比例滴入壳聚糖溶液中，调节PH为5.5，得到壳聚糖修饰的玉屏风多糖脂质体。壳聚糖表面带正电荷，脂质体脂膜表面带负电荷，通过电荷作用可以相互结合。经过壳聚糖修饰的玉屏风多糖脂质体提高了其消化稳定性，防止脂质体脂膜在胃部遭受严重破坏，使玉屏风多糖脂质体能够顺利到达肠道并吸收。
Description
技术领域
本发明涉及改变食品营养性质技术领域，特别涉及一种玉屏风多糖脂质体的制备方法。
背景技术
脂质体是由磷脂双分子层形成的、内部包含水相的封闭囊泡，具有缓释、细胞亲和、组织相容和靶向性等优点。然而，脂质体作为一种微粒分散体系，在贮藏过程中易氧化、絮凝，导致粒径变大和药物渗漏。特别是常规的玉屏风多糖脂质体，其在人工肠液稳定性较高，但在人工胃液中稳定性差，易造成药物渗漏。
发明内容
本发明针对上述问题，提供一种玉屏风多糖脂质体的制备方法。在制备玉屏风多糖脂质体的过程中增加了改性修饰脂质体的步骤，使制得的玉屏风多糖脂质体在人工胃液中的消化稳定性得到提升。
本发明进一步解决其技术问题所采用的技术方案如下：
一种玉屏风多糖脂质体的制备方法，包括以下步骤：
1)将大豆卵磷脂和胆固醇溶于氯仿，减压旋转蒸发，使溶液在容器内壁形成一层淡黄色脂质薄膜，随后加入***将薄膜溶解，加入玉屏风多糖和PH值7.4的磷酸盐缓冲液，进行超声处理充分混匀，随后减压抽吸至***挥发干净，即得玉屏风多糖脂质体基体；
2)将玉屏风多糖脂质体基体按照1:1的比例滴入壳聚糖溶液中，调节PH为5.5，得到壳聚糖修饰的玉屏风多糖脂质体。
进一步，步骤1所述大豆卵磷脂和胆固醇的质量比为(8～10):1。所述玉屏风多糖的添加质量为大豆卵磷脂的0.20～0.25倍。所述磷酸盐缓冲液的浓度为10mg/mL。
进一步，所述壳聚糖溶液的制备方式为：称取0.25g的壳聚糖粉末溶解在50ml体积分数为1％的乙酸溶液中，并充分搅拌使其溶解，过滤除渣。
本发明与现有技术相比具有优点：壳聚糖表面带正电荷，脂质体脂膜表面带负电荷，通过电荷作用可以相互结合。经过壳聚糖修饰的玉屏风多糖脂质体提高了其消化稳定性，防止脂质体脂膜在胃部遭受严重破坏，使玉屏风多糖脂质体能够顺利到达肠道并吸收。
附图说明
图1是玉屏风多糖脂质体修饰前和修饰后的渗透率变化折线图；
图2是壳聚糖的HPLC色谱图；
图3是修饰后玉屏风多糖脂质体的HPLC色谱图。
具体实施方式
下面通过具体实施例，对本发明的技术方案作进一步具体的说明，但是本发明并不限于这些实施例。
实施例1，制备经壳聚糖修饰的玉屏风多糖脂质体的方法。
脂质体基体的制备：采用逆向蒸发法，将大豆卵磷脂和胆固醇按8：1的质量比例溶于氯仿，采用旋转蒸发仪进行减压旋转蒸发，使圆底烧瓶内壁形成一层淡黄色脂质薄膜，随后加入适量的***将薄膜溶解，加入玉屏风多糖以及浓度为10mg/mL的磷酸盐缓冲液(PH＝7.4)，进行超声处理充分混匀，随后在减压旋转蒸发仪上减压抽吸至***挥发干净，即得乳白色得玉屏风多糖脂质体。所述玉屏风多糖与大豆卵磷脂的质量比为1：4。
壳聚糖溶液的制备：称取0.25g的壳聚糖粉末溶解在50ml体积分数为1％的乙酸溶液中，并充分搅拌使其溶解，过滤除渣。
壳聚糖修饰玉屏风多糖脂质体的制备：将玉屏风多糖脂质体按照1:1的比例滴入壳聚糖溶液中，调节PH为5.5，得到壳聚糖修饰的玉屏风多糖脂质体。
实施例2，验证壳聚糖修饰的玉屏风多糖脂质体在人工胃液中的稳定性。
人工胃液的配制：取浓度为1mol/ml的稀盐酸，加水稀释，将pH调至1.5。每100ml液体中加入1g胃蛋白酶，混匀，用0.2um的无菌滤头过滤待用。
分别取50ml人工胃液两份，各自精密吸取实施例1制得的脂质体溶液或未经修饰的脂质体溶液5ml加入，置于37℃磁力搅拌器中，分别于0、0.5、1、2、4、6、8h取样1ml于10ml容量瓶中，以蒸馏水定容，摇匀。将样品溶液用三氟乙酸在高压的情况下水解成单糖，然后进行PMP衍生化处理。结果如图1所示，
色谱条件；Wondasil C18(4.6×250nm，5微米)色谱柱，流动相为磷酸盐缓冲盐(PH6.9)-乙腈(85：15)，柱温为30℃；流速为1mL/min；进样量为20微升；检测波长为255nm。
玉屏风多糖在此色谱条件下会出现D-葡萄糖、D-半乳糖、D-***糖三种单糖的吸收峰，而壳聚糖水解的成D-葡萄糖，对其他两种单糖的吸收峰无影响。检测结果如图2和3所示，壳聚糖水解成单糖为D-葡萄糖，不影响D-半乳糖与D-***糖的峰面积。
壳聚糖表面带正电荷，脂质体脂膜表面带负电荷，通过电荷作用可以相互结合。经过壳聚糖修饰的玉屏风多糖脂质体提高了其消化稳定性，防止脂质体脂膜在胃部遭受严重破坏，使玉屏风多糖脂质体能够顺利到达肠道并吸收。
以上详细描述了本发明的优选实施方式，但是本发明不限于上述实施方式中的具体细节，在本发明的技术构思范围内，可以对本发明的技术方案进行多种简单变型，这些均属于本发明的保护范围。
Claims (6)
1.一种玉屏风多糖脂质体的制备方法，其特征在于，包括以下步骤：
1)将大豆卵磷脂和胆固醇溶于氯仿，减压旋转蒸发，使溶液在容器内壁形成一层淡黄色脂质薄膜，随后加入***将薄膜溶解，加入玉屏风多糖和PH值7.4的磷酸盐缓冲液，进行超声处理充分混匀，随后减压抽吸至***挥发干净，即得玉屏风多糖脂质体基体；
2)将玉屏风多糖脂质体基体按照1:1的比例滴入壳聚糖溶液中，调节PH为5.5，得到壳聚糖修饰的玉屏风多糖脂质体。
2.根据权利要求1所述的玉屏风多糖脂质体的制备方法，其特征在于，步骤1所述大豆卵磷脂和胆固醇的质量比为(8～10):1。
3.根据权利要求1所述的玉屏风多糖脂质体的制备方法，其特征在于，步骤1所述玉屏风多糖的添加质量为大豆卵磷脂的0.20～0.25倍。
4.根据权利要求1所述的玉屏风多糖脂质体的制备方法，其特征在于，步骤1所述磷酸盐缓冲液的浓度为10mg/mL。
5.根据权利要求1所述的玉屏风多糖脂质体的制备方法，其特征在于，所述壳聚糖溶液的制备方式为：称取0.25g的壳聚糖粉末溶解在50ml体积分数为1％的乙酸溶液中，并充分搅拌使其溶解，过滤除渣。
6.一种玉屏风多糖脂质体，其特征在于，由权利要求1～5任一项制备方法制得。
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