Preparation and Stability of Nanostructured Lipid Carriers of Phytosterols with Walnut Oil
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摘要: 本研究通过高压均质法采用核桃油为液体脂质制备用于封装、保护植物甾醇（Phytosterol，PS）的纳米结构脂质载体（Nanostructured lipid carriers，NLC）。以平均粒径、多分散指数、Zeta电位及包封率等为主要评价指标，对制备工艺参数及配方进行优化，同时对优化后的PS-NLC进行形貌观察及稳定性研究。通过正交试验确定制备PS-NLC的最佳比例为总脂质浓度10%，硬脂酸和核桃油的比例为2:3，大豆卵磷脂浓度为1.2%。制备得到的PS-NLC外观呈球形，粒径较小且分布均匀。PS-NLC的稳定性结果表明：PS-NLC在4 ℃下储藏28 d稳定性良好；在使用时可以在5~100倍之间进行稀释，具有良好的稀释稳定性；添加2%的蔗糖对PS-NLC的冻干保护效果最佳。本文利用核桃油作为NLC的壁材为植物甾醇提供了一个合适的脂质运载系统，可为食品工业构建PS-NLC提供技术支持。Abstract: Nanostructured lipid carriers (NLC) for the encapsulation and protection of phytosterol (PS) were prepared using walnut oil as liquid lipid by high-pressure homogenization method. The average particle size, polydispersity index, Zeta potential and encapsulation efficiency were used as the main evaluation indexes to optimize the preparation process parameters and formulations, and the morphology and stability of the optimized PS-NLC were investigated. The optimal ratio for the preparation of PS-NLC was determined by orthogonal tests as 10% of total lipid concentration, 2:3 of stearic acid and walnut oil, and 1.2% of soy lecithin. The prepared PS-NLC had a spherical appearance, small particle size and uniform distribution. The stability results of PS-NLC showed that PS-NLC was stable in storage at 4 ℃ for 28 days. It was diluted 5~100 times with good dilution stability, when used. And the addition of 2% sucrose had the best effect on the lyophilization protection of PS-NLC. Walnut oil was used as the wall material of NLC to provide a suitable lipid delivery system for phytosterols, which could provide technical support for the construction of PS-NLC in food industry.
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Key words:
- phytosterol /
- nanostructured lipid carriers /
- walnut oil /
- soy lecithin /
- stearic acid
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图 1 不同高压均质条件对PS-NLC的粒径及PDI的影响
注：同一指标字母不同表示差异有统计学意义（P<0.05）。
Figure 1. Effects of different high pressure homogenization conditions on particle size and PDI of PS-NLC
图 2 不同种类的表面活性剂对PS-NLC的粒径和Zeta电位影响
注：字母不同表示差异有统计学意义（P<0.05）。
Figure 2. Effects of different types of surfactants on particle size and Zeta potential of PS-NLC
图 4 不同处理条件PS-NLC的粒径和PDI变化
注：（A）稀释倍数，（B）蔗糖浓度，同一指标字母不同表示差异有统计学意义（P<0.05）。
Figure 4. Changes in particle size and PDI of PS-NLC under different experimental conditions
表 1 正交试验因素水平设计
Table 1. Orthogonal assay factors and levels
水平 因素 A总脂质浓度（%） B固液脂质量比 C表面活性剂浓度（%） 1 7.5 4:1 1.2 2 10 3:2 1.4 3 12.5 2:3 1.6 
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表 2 四种不同固体脂制备的PS-NLC的粒径、PDI及包封率
Table 2. Particle size, PDI and encapsulation ratio of PS-NLC prepared from four different solid lipids
固体脂质 粒径（nm） PDI 包封率（%） 棕榈酸 525.43±4.631b 0.357±0.013b 77.10±0.116d 硬脂酸 302.90±0.794d 0.238±0.011c 91.09±0.115b 山嵛酸甘油酯 740.90±7.503a 0.206±0.007d 85.63±0.364c 单硬脂酸甘油酯 344.47±4.801c 0.592±0.021a 92.29±0.236a 注：每一列上标字母不同表示均值差异显著（P<0.05），字母相同表示均值差异不显著（P>0.05）；表4同。
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表 3 L9（34）正交试验设计及结果
Table 3. L9(34) orthogonal experiment design and results
试验号 总脂质浓度（%） 固液脂质量比 表面活性剂浓度（%） 包封率（%） 1 7.5 4:1 1.2 85.85 2 7.5 3:2 1.4 88.68 3 7.5 2:3 1.6 84.58 4 10 4:1 1.4 87.24 5 10 3:2 1.6 86.43 6 10 2:3 1.2 93.97 7 12.5 4:1 1.6 84.39 8 12.5 3:2 1.2 91.77 9 12.5 2:3 1.4 90.88 K1 86.370 85.827 90.530 − K2 89.213 88.960 88.933 − K3 89.013 89.810 85.133 − R 2.843 3.983 5.937 −
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表 4 4 ℃下储存28 d的PS-NLC的粒径、PDI和Zeta电位
Table 4. Particle size, PDI and Zeta potential of PS-NLC stored at 4 ℃ for 28 days
储藏时间（d） 粒径（nm） PDI Zeta电位（mV） 0 313.23±0.666d 0.248±0.003d −35.23±0.057b 7 339.80±1.646a 0.253±0.005c −34.67±0.208a 14 322.63±2.843c 0.236±0.003e −35.30±0.361b 21 319.73±1.193c 0.267±0.003b −35.83±0.208c 28 327.93±2.409b 0.288±0.005a −36.10±0.264c
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