CN108338356B - 一种壳聚糖包裹的甘露糖赤藓糖醇脂脂质体及其制备方法 - Google Patents
一种壳聚糖包裹的甘露糖赤藓糖醇脂脂质体及其制备方法 Download PDFInfo
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- CN108338356B CN108338356B CN201810042262.2A CN201810042262A CN108338356B CN 108338356 B CN108338356 B CN 108338356B CN 201810042262 A CN201810042262 A CN 201810042262A CN 108338356 B CN108338356 B CN 108338356B
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Abstract
本发明公开了一种壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,包括脂质体载体,包封于脂质体载体内的脂溶性活性物质和包裹于脂质体载体外的壳聚糖。本发明以甘露糖赤藓糖醇脂和大豆卵磷脂自组装制备脂质体载体,用于包封脂溶性活性物质,并以壳聚糖包裹在外层,得到的脂质体具有较好的生物相容性、粘膜粘附性和稳定性,安全性较高,对脂溶性活性物质具有较高的包封率,且能增强其抗氧化活性。所用的材料安全无毒,载体的制备过程无需借助其他助剂,其方法简单可行,成本低。
Description
技术领域
本发明涉及一种壳聚糖包裹的甘露糖赤藓糖醇脂脂质体及其制备方法,属于生物材料与食品添加剂领域。
背景技术
目前,食品被赋予了更广泛的定义,其中除了满足人们的基本能量和营养需求外,还应包括增强身心健康的功能效应。然而,生物活性物质通常对氧化,消化或其他反应敏感。因此,对生物活性化物质的递送系统的开发正在变得原来越重要。
脂质体由于其能够阿伯在物质的广泛性,被认为是一种十分有前景的递送系统,并广泛用于食品,化妆品,生物和制药研究和工业。在药物领域,脂质体可降低治疗剂的毒性,靶向针对特定的器官或部位,并增加药物对某些组织的渗透。此外,脂质体在生物相容性,无免疫原性,大分子负载能力和加载水溶性和脂溶性化合物方面优于其它体系。尤其对于疏水剂,脂质体中的包封可以显著增加其水溶性。在食品研究和工业领域,脂质体已被用于传递风味物质,酶,营养物质和功能成分,还被用于防止食物的腐败。
生物表面活性剂是由多种微生物产生的两亲性化合物,已被应用于食品,化妆品,制药,石化,环保等多个领域。甘露糖赤藓糖醇脂是含亲水部分4-O-β-D-吡喃甘露糖基-赤藓糖醇和疏水部分脂肪酸和乙酰基的糖脂生物表面活性剂。甘露糖赤藓糖醇酯分为具有不同乙酰基和脂肪酸的四种化学结构,即A,B,C和D型。甘露糖赤藓糖醇脂的可生物降解性,稳定性和乳化能力优异。有趣的是,甘露糖赤藓糖醇脂能够诱导人类早幼粒细胞白血病细胞系和大鼠嗜铬细胞瘤的细胞凋亡和细胞分化。另外,甘露糖赤藓糖醇脂具有抗氧化和抗菌作用。特别的,含有甘露糖赤藓糖醇脂的脂质体能提高基因转染效率,增强细胞融合和膜融合。有文献报道,人造卵磷脂DLPC和A型甘露糖赤藓糖醇脂可以组装脂质体。
使用生物高分子物质包裹脂质体不仅可以获得更高的稳定性和更长的贮藏期限,而且还可以使脂质体不易在胃酸性环境和胃及小肠中的酶消化作用下受损。由β-1,4-连接的2-乙酰氨基-2-脱氧-D-吡喃葡萄糖和2-氨基-2-脱氧-D-吡喃葡萄糖单元组成的壳聚糖是可生物降解的,生物相容的和粘膜粘附的生物高分子物质之一。
发明内容
本发明提供了一种壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,具有较好的生物相容性、粘膜粘附性和稳定性,安全性较高,对脂溶性活性物质具有较高的包封率,且能增强其抗氧化活性。
一种壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,包括脂质体载体,包封于脂质体载体内的脂溶性活性物质和包裹于脂质体载体外的壳聚糖;
其中,所述脂质体载体由甘露糖赤藓糖醇脂和大豆卵磷脂在水溶液中自组装制成。
壳聚糖可生物降解,生物相容性和粘膜粘附性较好,经壳聚糖包裹的脂质体可延长脂质体在胃肠道中的滞留并改善脂质体对黏膜的渗透,促进吸收。壳聚糖包裹还可提高脂质体的稳定性、包封率和抗氧化活性。
甘露糖赤藓糖醇脂(MEL-A)是一种天然生物表面活性剂,既有优良的表面性能,又易于在环境中被生物降解;大豆卵磷脂为混合磷脂(主要含有卵磷脂、脑磷脂、肌醇磷脂、磷酯酰丝氨酸、磷脂酸及其他磷脂等),作为一种常用的食品添加剂,相比合成磷脂(DLPC),大豆卵磷脂安全性更好,能够大量应用于食品中,且其成本较低,适用于食品体系中递送活性物质的脂质体制备。MEL-A与大豆卵磷脂自组装得到的脂载体,粒径分布更加均匀,稳定性更好,能作为许多疏水性及不稳定性活性成分的载体,解决脂溶性活性物质的水溶性与载运难题。
作为优选,所述甘露糖赤藓糖醇脂的主要分子量分布在550-690。
作为优选,所述甘露糖赤藓糖醇脂的纯度在95%以上。
所述甘露糖赤藓糖醇脂可以为Ustligo maydis属和Pseudazyma属菌种发酵得到的发酵液中分离纯化得到的。
作为优选,所述大豆卵磷脂的纯度在98%以上。
所述甘露糖赤藓糖醇脂和大豆卵磷脂的质量比可以为1∶10-10∶1;更优选为3∶7-5∶5,最优选为6∶4,自组装效果更好,形成的脂质体载体的多分散指数(PDI)值低于0.469,粒径分布更加均匀,稳定性较好,对活性物质包裹性较好。
所述甘露糖赤藓糖醇脂和大豆卵磷脂在壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中的总浓度可以为0.25-1.0mg/mL,优选为0.5mg/mL。
所述脂溶性活性物质可以为白桦脂酸、维生素E、α-亚麻酸中的至少一种。
当脂溶性活性物质为白桦脂酸时,所述白桦脂酸在壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中的用量为0.03-0.05mg/mL,优选为0.04mg/mL,载药量合适,包封效果较好,包封率可达60-80%。
所述壳聚糖可以为市售的商品壳聚糖,购于青岛裕达世纪经贸有限公司,为从深海阿拉斯加雪蟹壳中提取得到的。
所述壳聚糖也可以为真菌壳聚糖,为从野生柴达木大肥菇(Agaricus bisporus(Lange)Sing.Chaidam)的子实体中提取、分离得到。
所述真菌壳聚糖可以通过如下方法制备:将野生柴达木大肥菇(Agaricusbisporus(Lange)Sing.Chaidam)子实体冷冻干燥、粉碎,然后采用碱法提制,得到所述的真菌壳聚糖。
所述壳聚糖在壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中的用量为0.05-3mg/mL,优选为2mg/mL,包裹效果最好,包裹后脂质体稳定性得到极大提高,对脂溶性活性物质白桦脂酸的包封率最高,并赋予脂质体体系较强的抗氧化活性。
本发明还提供了上述壳聚糖包裹的甘露糖赤藓糖醇脂脂质体的制备方法,包括:
(1)将甘露糖赤藓糖醇脂、大豆卵磷脂和脂溶性活性物质分别溶于氯仿得到溶液,将三种溶液混合均匀,在减压条件下旋转蒸发成膜,避光静置除去有机溶剂;然后加入蒸馏水,混匀,超声,静置,得到脂质体悬浮液;
(2)将壳聚糖溶于乙酸得到壳聚糖溶液,然后以边滴加边搅拌的方式加入到步骤(1)制得的脂质体悬浮液中,室温搅拌0.5-1.5h,然后于4℃下孵育过夜,得到壳聚糖包裹的甘露糖赤藓糖醇脂脂质体。
步骤(1)中,甘露糖赤藓糖醇脂和大豆卵磷脂的质量比优选为1∶10-10∶1;更优选为3∶7-5∶5;最优选为4∶6。
脂质体悬浮液中,甘露糖赤藓糖醇脂和大豆卵磷脂的总浓度为0.5-2.0mg/mL,优选为1.0mg/mL。
所述脂溶性活性物质可以为白桦脂酸。脂质体悬浮液中,所述白桦脂酸的浓度为0.06-0.1mg/mL,优选为0.08mg/mL。
步骤(2)中,所述壳聚糖在壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中的用量为0.05-3mg/mL,优选为2mg/mL。
本发明以甘露糖赤藓糖醇脂和大豆卵磷脂自组装制备脂质体载体,用于包封脂溶性活性物质,并以壳聚糖包裹在外层,得到的脂质体具有较好的生物相容性、粘膜粘附性和稳定性,用于包封白桦脂酸时,对白桦脂酸的包封率达到60-80%,且增强了其抗氧化活性。所用的材料安全无毒,载体的制备过程利用了甘露糖赤藓糖醇脂MEL-A具有的自组装性质与大豆卵磷脂复配而成,无需借助其他助剂,其方法简单可行,成本低。
附图说明
图1中,图(a)为傅里叶变换红外光谱图,图(b)为X射线衍射图,图(c)为差式热量扫描结果;其中,1为来自蟹的市售壳聚糖,2为真菌壳聚糖,3为来自真菌几丁质脱乙酰化的壳聚糖,4为真菌几丁质。
图2为透射电镜下壳聚糖包裹的甘露糖赤藓糖醇脂脂质体的形态;其中,图(a)和图(c)为10000和50000倍放大的未经壳聚糖包裹的脂质体,图(b)和图(d)为10000和50000倍放大的0.4%壳聚糖包裹的脂质体。
图3为代表性的脂质体粒径分布图;其中,图(a)为不载药脂质体,图(b)为包载白桦脂酸的0.1%壳聚糖包裹的脂质体。
图4为脂质体的差事热量扫描结果。
图5为脂质体的抗氧化结果;其中,图(a)为DPPH清除率结果;图(b)为ABTS清除率结果。
具体实施方式
下述实施例中所使用的实验方法如无特殊说明,均为常规方法。
下述实施例中所采用的材料、试剂扥,如无特殊说明,均可从商业途径得到。
下述实施例中,实验试剂来源:
商品壳聚糖来自青岛裕达世纪经贸有限公司,原料为深海阿拉斯加雪蟹壳。大豆卵磷脂(>98%)购自阿拉丁(中国上海)。真菌壳聚糖提取自野生柴达木大肥菇(Agaricusbisporus(Lange)Sing.Chaidam)子实体,该子实体从青海柴达木盆地采集分离得到。
实施例1脂质体的制备
一、脂质体材料制备
1、甘露糖赤藓糖醇脂的生产与纯化
甘露糖赤藓糖醇脂由蚜虫拟酵母(Pseudozyma aphidis)DSM70725产生。A型甘露糖赤藓糖醇脂(MEL-A)的生产和纯化方法:首先将蚜虫拟酵母接种到含有3%酵母提取物,3%麦芽提取物,10%葡萄糖和5%蛋白胨的活化培养基中,并在28℃和180rpm下培养36小时。活化后,将1mL菌液培养物接种到由NaNO3 3g/L,MgSO4·7H2O 0.3g/L,KH2PO4 0.3g/L,酵母提取物1g/L,葡萄糖40g/L和蒸馏水组成的种子培养基。在28℃和180rpm培养2天后,将种子培养物离心并用生理盐水洗涤两次。将得到的细胞接种于含80mL/L大豆油,MgSO4·7H2O0.3g/L,KH2PO4 0.3g/L,NaNO33g/L,酵母提取物1g/L的发酵培养基中。发酵过程在28℃,180rpm下进行7天。发酵结束后,加入相同体积的乙酸乙酯并充分震荡萃取甘露糖赤藓糖醇脂。
将混合物以3,800rpm离心15分钟,分离有机层并减压蒸发。用甲醇:环己烷(1∶1,v/v)萃取两次以除去剩余的油和脂肪酸后,用硅胶柱从粗制MEL中纯化MEL-A,并通过薄层色谱(硅胶GF254,三氯甲烷∶甲醇∶水=65∶15∶2,v/v)检验产物。
2、真菌壳聚糖的提取
将野生柴达木大肥菇(Agaricus bisporus(Lange)Sing.Chaidam)子实体冷冻干燥。冻干后使用粉碎机充分粉碎至60目。将一定量的子实体粉末加入到1mol/L的NaOH溶液(1∶30,w/v)中,并在90℃下搅拌2h。将悬浮液以5000g离心20分钟,并用蒸馏水洗涤至中性pH。几丁质在上清液中,壳聚糖在残留物中。沉淀用90倍体积的5%乙酸处理,振荡3h。用4mol/L NaOH将酸溶部分调节至pH10。离心收集壳聚糖,分别用蒸馏水,95%乙醇和丙酮洗涤。将充分洗净的沉淀冻干获得真菌壳聚糖。真菌几丁质在60℃下用47%的NaOH作用2小时。用热水洗涤沉淀至中性pH并冻干。
3、壳聚糖的表征
傅里叶变换红外(FTIR)光谱:
傅里叶变换红外光谱在FTIR光谱仪(NicoletAvatar 370,Madison,USA)上从4000-400cm-1进行。商品壳聚糖,真菌壳聚糖,真菌几丁质和真菌几丁质脱乙酰壳聚糖通过KBr小片法制备。
从图1a可以看出,商品壳聚糖,真菌壳聚糖和真菌几丁质具有相似的红外光谱。多糖的结构可以通过光谱中的特征峰显示。以3414cm-1为中心的宽吸收带归属于轴向NH和OH键。CH键导致2874cm-1的峰。以1653,1597和1379cm-1为中心的特征峰归属于酰胺I,酰胺II和酰胺III谱带,四条曲线上这三个峰的强度差异是由于壳聚糖的各种脱乙酰度引起的。CO和COC键导致以1092cm-1为中心的信号。除了这些提到的峰外,还有一个以897cm-1为中心的属于CH糖苷键的峰。
脱乙酰度(DD):
如前所述,乙酰化程度(DA)由通过傅里叶变换红外光谱获得的A1655和A3450的吸光度比确定。DA由以下等式计算:
DA(%)=(A1655/A3450)*100/1.33
然后通过以下公式计算DD:
DD(%)=100-DA
发现商品壳聚糖,真菌壳聚糖,几丁质脱乙酰壳聚糖和真菌几丁质的DD值分别为68.61,66.35,64.08和58.03%。结果表明,真菌壳聚糖与市售壳聚糖具有相似的DD。
X射线衍射:
使用PANalytical X-pert粉末衍射仪(PANalytical,荷兰)收集X射线衍射数据。样品在40KV和40mA下从2V到5V扫描。
X射线衍射图谱如图1b所示。几丁质和壳聚糖四种样品的衍射角均在之间。显然,它们在10.5和20.0o处都有相似的特征峰。这一结果表明,从真菌中提取的产物都是壳聚糖或几丁质。但是,这个峰值强度不一样。商品壳聚糖在20o左右峰值强度最高,远高于其他样品,说明真菌壳聚糖的结晶度较低。
差示扫描量热法(DSC)测量:
在CEC-130263F DSC(Mettler Toledo,瑞士)上测量市售壳聚糖和真菌壳聚糖的热性质。将一定量的样品装入小铝坩埚中,然后盖上盖子。在20~500℃的加热过程中,以10℃/分钟的速率收集DSC数据,干燥氮气流保护。
关于热性能,采用差式热量扫描分析壳聚糖和真菌壳聚糖。图1c显示了蟹壳聚糖和真菌壳聚糖的DSC曲线,显示它们具有相似的DSC模式。吸热峰出现在140℃附近,归因于聚合物中含水量的损失。聚合物链的分解导致在310℃附近观察到放热峰。
分子量测定:
样品的分子量由高效凝胶色谱法测定。该系统由配备有TSK-GEL G2000SWXL(TOSOH,日本)柱(7.8mm×300mm)和2414型折射率检测器(RID)(Waters Co.)的Waters2695装置美国)。使用Na2SO4(0.2mol/L)作为洗脱液,流速为1.0mL/min。用0.22μm注射器膜过滤1.0mg/mL样品溶液,并注射10μL样品。柱温固定在35℃。T系列葡聚糖标准被应用于校准。根据标准曲线计算样品的分子量。
提取的真菌壳聚糖的平均分子量为37354.2Da。
4、壳聚糖包裹的甘露糖赤藓糖醇脂脂质体制备
脂质体通过薄膜方法制备,具体为:
(1)将甘露糖赤藓糖醇脂(MEL-A)、大豆卵磷脂、白桦脂酸分别溶于氯仿得到溶液,将三种溶液混合均匀,在减压(35℃,50rpm)下旋转蒸发成膜,避光静置一天至残留痕量有机试剂完全挥发;然后加入蒸馏水进行水合,并进行振荡(180rpm,30min)、超声(30min),静置,得到脂质体悬浮液;
其中,MEL-A和大豆卵磷脂的质量比为4∶6;脂质体悬浮液中,MEL-A和大豆卵磷脂的总浓度为1.0mg/mL,白桦脂酸的浓度为0.08mg/mL;
(2)将壳聚糖溶于浓度为1%(v/v)的乙酸中并磁力搅拌过夜,得到壳聚糖溶液;然后在磁力搅拌下,以边滴加边搅拌的方式将不同浓度的壳聚糖溶液加入到等体积的步骤(1)制得的脂质体悬浮液中,室温搅拌1h,然后于4℃下孵育过夜,促进壳聚糖涂层的完整形成,得到壳聚糖包裹的甘露糖赤藓糖醇脂脂质体。
其中,壳聚糖溶液的浓度分别为0.01、0.05、0.1、0.2、0.4和0.6%(w/v)。
在制得的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中,MEL-A和大豆卵磷脂的总浓度相当于0.5mg/mL,白桦脂酸的用量相当于0.04mg/mL,壳聚糖的用量相当于0.05-3mg/mL。
实施例2脂质体的表征
一、脂质体的物理性质表征
粒径分布和zeta电位:
通过动态光散射(DLS)在633nm波长处适应Zetasizer Nano ZS90(MalvernInstruments Ltd,Malvern,Worcestershire,UK)测定脂质体的粒径分布、分散指数和zeta电位。温度设定为25℃,测定角度为90°。还进行差式热量扫描。样品从-40℃加热到100℃,速度为5℃/min。
包封率测定:
将1mL脂质体混悬液以1000rpm离心10分钟以除去游离白桦脂酸。然后将0.5mL上清液以12000rpm离心20分钟以获得所有脂质体结构的沉淀。用甲醇和氯仿(1∶1,v/v)释放白桦脂酸,溶液用超声处理2分钟。有机溶剂在37℃和50rpm旋转蒸发下蒸干。然后,加入甲醇以溶解残余物用于白桦脂酸测定。通过反相HPLC(RP-HPLC)方法测定白桦脂酸浓度。使用的柱是反相对称C18(250mm×4.6mm i.d.,4μL;Waters)。乙腈:水为91:9(v/v)作为流动相,流速1.0mL/min,30℃。检测波长固定在210nm。以下等式用于计算脂质体的白桦脂酸包封率:
EE(%)=脂质体中的白桦脂酸/(脂质体中的白桦脂酸+未被包载的白桦脂酸)*100%
形态观察:
用JEM-1200EX透射电子显微镜(JEOL,日本)观察真菌包被的脂质体的形态。将具有薄膜的铜网浸入样品溶液中,然后用乙酸铀酰染色。使用薄纸小心地吸干多余的液体,而不接触铜网。然后在120KV的加速电压下分析样品。
表1列出了不同真菌壳聚糖浓度对平均粒径,多分散吸水和zeta电位的影响。图2a和2b显示了真菌壳聚糖包裹的和未包裹的脂质体的典型粒径分布。由MEL-A和大豆卵磷脂以4∶6组成的空脂质体具有最小的粒径,平均直径为286.7±2.0nm(图2a)。包载白桦脂酸后,平均粒径略有增加,达到292.5±9.8nm。真菌壳聚糖包裹后,随着壳聚糖浓度的增加,平均粒径从303.6±1.6nm变为706.1±7.5nm。脂质体直径的增长可以通过带正电荷的壳聚糖和带负电荷的脂质体脂质成分之间的离子相互作用来解释,这导致形成壳聚糖包裹层脂质体。
多分散指数(PDI)揭示了所测样品的尺寸分布。PDI值是从0到1的数字,较高的分散系数表明样品具有宽范围的粒度,相反,较低的PDI值表明样品尺寸更均匀。真菌壳聚糖涂布或未涂布的脂质体的PDI值均低于0.469,表明脂质体的粒径处于相当均匀的状态。
真菌壳聚糖之间的电子相互作用也有助于脂质体的稳定性。Zeta电位是可以在一定程度上代表脂质体的稳定性的参数。高zeta电位(负或正)表示样品中的颗粒具有强排斥性,并且它们聚集的趋势较弱。本实施例中所有脂质体的Zeta电位如表1所示。未经壳聚糖包裹的空白脂质体和未经壳聚糖包裹的含有白桦脂酸的脂质体的zeta电位分别为16.6±0.5mV和26.0±0.8mV。真菌壳聚糖包被后,zeta电位值变为正值,并显著增加。随着壳聚糖浓度的增加,zeta电位先增大后减小。在0.1%的壳聚糖浓度条件下获得最高的zeta电位。
表1含有白桦脂酸的真菌壳聚糖包裹的甘露糖赤藓糖醇脂脂质体的平均粒径,多分散系数和Zeta电位
不同上标字母表示不同处理间差异显着(p<0.05)。
二、抗氧化效果评估
1,1-二苯基-2-picrylhydrazyl(DPPH)自由基清除活性测定:
将1mL溶于95%乙醇的0.1mmol/L DPPH溶液与4mL样品溶液混合。将混合物在黑暗环境中在室温下孵育30分钟,然后在517nm处测量吸光度。每个样品三个平行。样品的清除率由以下公式计算:
DPPH清除率(%)=(空白吸光值-样品吸光值)/空白吸光值
白桦脂酸具有多种生物活性,但其自由基清除能力非常弱(图5a)。白桦脂酸浓度设定为0.4mg/mL,接近具有最高包封率的脂质体的白桦脂酸浓度。如图5a所示,白桦脂酸具有较弱的DPPH自由基清除能力。未经壳聚糖包裹的脂质体具有比白桦脂酸更高的自由基清除率。这可能是由于MEL-A的抗氧化活性。壳聚糖包裹后,自由基清除率随着壳聚糖浓度的增加而增加。自由基清除率的提高可能是由于壳聚糖的抗氧化活性。当壳聚糖浓度达到0.6%时,DPPH自由基清除率是白桦脂酸的20倍以上。
用ABTS法测定总抗氧化能力:
使用ABTS试剂盒(Beyotime生物技术研究所,中国)测定ABTS抗氧化活性。ABTS储备溶液是通过将100μLABTS溶液和100μL氧化剂溶液混合并在室温下在黑暗环境中反应12-16小时而制备的。在使用之前,将80%(v/v)乙醇加入到储备液中,直到734nm处的吸光度变为0.7±0.5。将200mL ABTS工作溶液与10mL样品溶液混合,并在室温下孵育4分钟。然后测量734nm处的吸光度。ABTS抗氧化活性计算如下:
ABTS自由基清除能力(%)=(空白吸光值-样品吸光值)/空白吸光值
白桦脂酸ABTS自由基清除率较低,约为4%,这与DPPH实验结果一致。尽管未经壳聚糖包裹的脂质体和壳聚糖包被的脂质体在0.01,0.05和0.1%(w/v)的壳聚糖引起抗氧化活性下降,但在0.2,0.4和0.6%(w/v)的较高壳聚糖浓度导致相当或显着更有效的ABTS自由基清除能力(p<0.05)。在DPPH和ABTS实验中观察到的抗氧化作用的增强可能归因于真菌壳聚糖中的氨基和乙酰氨基的抗氧化能力。用0.6%壳聚糖包裹后,脂质体的ABTS自由基清除率比白桦脂酸高2倍以上。
Claims (9)
1.一种壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,包括脂质体载体,包封于脂质体载体内的脂溶性活性物质和包裹于脂质体载体外的壳聚糖;
所述脂溶性活性物质为白桦脂酸;所述白桦脂酸在壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中的用量为0.03-0.05 mg/mL;
其中,所述脂质体载体由甘露糖赤藓糖醇脂和大豆卵磷脂在水溶液中自组装制成。
2.如权利要求1所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,所述甘露糖赤藓糖醇脂的主要分子量分布在550-690。
3.如权利要求1所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,所述大豆卵磷脂的纯度在95%以上。
4.如权利要求1所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,所述甘露糖赤藓糖醇脂和大豆卵磷脂的质量比为1:10-10:1。
5.如权利要求1所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,所述甘露糖赤藓糖醇脂和大豆卵磷脂在壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中的总浓度为0.25-1.0 mg/mL。
6.如权利要求1所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,所述脂溶性活性物质为白桦脂酸、维生素E、α-亚麻酸中的至少一种。
7.如权利要求1所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,所述壳聚糖为真菌壳聚糖,为从野生柴达木大肥菇(Agaricus bisporus (Lange) Sing. Chaidam)的子实体中提取、分离得到。
8.如权利要求1所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体,其特征在于,所述壳聚糖在壳聚糖包裹的甘露糖赤藓糖醇脂脂质体中的用量为0.05-3 mg/mL。
9.如权利要求1-8任一项所述的壳聚糖包裹的甘露糖赤藓糖醇脂脂质体的制备方法,其特征在于,包括:
(1)将甘露糖赤藓糖醇脂、大豆卵磷脂和脂溶性活性物质分别溶于氯仿得到溶液,将三种溶液混合均匀,在减压条件下旋转蒸发成膜,避光静置除去有机溶剂;然后加入蒸馏水,混匀,超声,静置,得到脂质体悬浮液;
(2)将壳聚糖溶于乙酸得到壳聚糖溶液,然后以边滴加边搅拌的方式加入到步骤(1)制得的脂质体悬浮液中,室温搅拌0.5-1.5 h,然后于4℃下孵育过夜,得到壳聚糖包裹的甘露糖赤藓糖醇脂脂质体。
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