CN104012991A - 乙醇注入-动态高压微射流制备丁香酚纳米脂质体的方法 - Google Patents
乙醇注入-动态高压微射流制备丁香酚纳米脂质体的方法 Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 13
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 22
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 claims abstract description 11
- 239000000787 lecithin Substances 0.000 claims abstract description 11
- 235000010445 lecithin Nutrition 0.000 claims abstract description 11
- 229940067606 lecithin Drugs 0.000 claims abstract description 11
- 239000002502 liposome Substances 0.000 claims description 61
- 235000019441 ethanol Nutrition 0.000 claims description 13
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 11
- 229920000053 polysorbate 80 Polymers 0.000 claims description 11
- 235000012000 cholesterol Nutrition 0.000 claims description 10
- 239000000839 emulsion Substances 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 229940107161 cholesterol Drugs 0.000 claims description 4
- 238000003756 stirring Methods 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 abstract description 3
- HVYWMOMLDIMFJA-UHFFFAOYSA-N 3-cholesterol Natural products C1C=C2CC(O)CCC2(C)C2C1C1CCC(C(C)CCCC(C)C)C1(C)CC2 HVYWMOMLDIMFJA-UHFFFAOYSA-N 0.000 abstract 1
- 238000005538 encapsulation Methods 0.000 abstract 1
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- 239000002994 raw material Substances 0.000 abstract 1
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- 239000000725 suspension Substances 0.000 description 8
- 238000000338 in vitro Methods 0.000 description 5
- 239000004530 micro-emulsion Substances 0.000 description 5
- KEQXNNJHMWSZHK-UHFFFAOYSA-L 1,3,2,4$l^{2}-dioxathiaplumbetane 2,2-dioxide Chemical compound [Pb+2].[O-]S([O-])(=O)=O KEQXNNJHMWSZHK-UHFFFAOYSA-L 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
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- 102000004190 Enzymes Human genes 0.000 description 1
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23P—SHAPING OR WORKING OF FOODSTUFFS, NOT FULLY COVERED BY A SINGLE OTHER SUBCLASS
- A23P10/00—Shaping or working of foodstuffs characterised by the products
- A23P10/30—Encapsulation of particles, e.g. foodstuff additives
- A23P10/35—Encapsulation of particles, e.g. foodstuff additives with oils, lipids, monoglycerides or diglycerides
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Abstract
一种乙醇注入-动态高压微射流制备丁香酚纳米脂质体的方法,是以丁香酚、卵磷脂、胆固醇和吐温为原料,采用乙醇注入-动态高压微射流法制备得到纳米脂质体。本发明制备的脂质体平均粒径为58.6±3.4nm,分布系数为0.381±0.053,包封率达到了59.2%±4.7%,它具有缓释性,24h后有76.6%的丁香酚释放出来。
Description
技术领域
本发明涉及涉及水溶性药物纳米脂质体的制备方法,具体涉及一种乙醇注入-动态高压微射流制备丁香酚纳米脂质体的方法。
背景技术
丁香酚具有消炎、止痛、抗氧化、抑菌等功能,是保健食品、药品等的潜在使用对象,但它较差的水溶性限制了其在食品中的应用。脂质体(liposome)是由磷脂双分子层形成的内部包含水相的封闭囊泡。研究表明,脂质体具有缓释性、细胞亲和性、组织相容性和靶向性等优点,已成功应用于生物医药、化工农业等领域。脂质体用于包裹营养素、酶、食品添加剂、食品抗菌剂等食品运载体系显示出诱人前景。利用脂质体技术将丁香酚包封起来,可以克服丁香酚上述缺点。乙醇注入-动态高压微射流法制备的脂质体可以克服粗脂质体粒径大,不稳定等缺点,同时乙醇注入法突破了薄膜法洗膜难和小批量生产的障碍。
发明内容
本发明的目的是为优化丁香酚脂质体的制备工艺,提高脂质体的稳定性,评价丁香酚脂质体的抗菌活性、储藏稳定性等,为丁香酚脂质体的开发提供一种乙醇注入-动态高压微射流制备丁香酚纳米脂质体的方法。
本发明所述方法包括:
(1)丁香酚纳米脂质体的组分丁香酚、卵磷脂、胆固醇和吐温-80、pH=7.4浓度为 0.05M/L的磷酸盐缓冲溶液与无水乙醇的质量体积比为:在100mL pH=7.4浓度为0.05M/L的磷酸盐缓冲溶液中:丁香酚 0.38-1.16g、卵磷脂 2.32-6.99g、胆固醇 0.35-1.04g、吐温-80 0.52-1.55g、 无水乙醇 25-35mL;
(2)按上述重量百分比分别称取丁香酚、卵磷脂、胆固醇和吐温-80,在40-50℃条件下,溶于无水乙醇;
(3)将步骤(2)中所得溶液缓慢注入到pH=7.4浓度为0.05M/L的磷酸盐缓冲溶液中,在搅拌的条件下将形成乳液;
(4)将步骤(3)中所得乳液移入圆底瓶,置于真空旋转蒸发仪上,在50℃水浴条件下除去乙醇,用磷酸盐缓冲溶液定溶,得到丁香酚粗脂质体;
(5)用动态高压微射流处理步骤(4)得到的粗脂质体,压力为120MPa,处理1次,即得丁香酚纳米脂质体。
本发明的有益效果是:
(1)包封率:乙醇注入-动态高压微射流法制备的丁香酚纳米脂质体的具有比较高的包封率,包封率达到59.2%±4.7。
(2)粒径:乙醇注入-动态高压微射流法制备的丁香酚纳米脂质体的平均粒径为58.6±3.4nm,分布系数为0.381±0.053。
(3)微观形貌图:乙醇注入-动态高压微射流法制备的丁香酚纳米脂质体,其透射电镜图如图2所示,脂质体成球形,大小及分布都比较均匀。
(4)乙醇注入-动态高压微射流法制备的丁香酚纳米脂质体具有缓释性,24h后有76.6%的丁香酚释放出来,其释放曲线如图3所示。
(5)储藏稳定性:分别将制备好的丁香酚纳米脂质体与丁香酚微乳(将丁香酚和吐温-80溶于PBS溶液中,持续搅拌)置于4℃和室温环境中,每隔一周测其粒径分布及包封率。如图4所示,丁香酚微乳有明显的油状沉淀,而丁香酚纳米脂质体则较为稳定。而图5所展示的平均粒径及包封率变化也证明丁香酚纳米脂质体在储藏期间较为稳定。
附图说明
图1 为乙醇注入-动态高压微射流法制备丁香酚纳米脂质体的工艺路线图;
图2为丁香酚纳米脂质体透射电镜图;
图3为丁香酚纳米脂质体及丁香酚微乳的体外释放曲线;
图4为丁香酚纳米脂质体及丁香酚微乳在4℃及室温环境中稳定性观察图;
图5为丁香酚纳米脂质体及丁香酚微乳在4℃及室温环境中粒径及包封率随时间的变化。
具体实施方式
实施例1
称取48.5g卵磷脂、6.6g胆固醇、12.2g吐温-80和8.2g丁香酚,完全溶解于0.32L无水乙醇中,将乙醇溶液缓慢注入到1 L pH7.4、浓度为0.05M的磷酸盐缓冲溶液中,形成乳液,然后将其转移到到圆底瓶中,在50℃水浴条件下真空旋转除去无水乙醇,形成均匀的悬浊液即为粗脂质体,形成的均匀悬浊液即为粗脂质体。将粗脂质体加入到动态高压微射流(DHPM)中,于120MPa条件下微流化处理1次,即制备得到丁香酚纳米脂质体。制得的纳米脂质体为较透明乳白色溶液。制得的丁香酚纳米脂质体的包封率为60.1%,平均粒径为60.4nm ,分布系数为0..332,在体外释放试验中,经24h释放后有78%的丁香酚释放出来。
实施例2
称取47.9g卵磷脂、6.1g胆固醇、12.2g吐温-80和8.0g丁香酚,完全溶解于0.28L无水乙醇中,将乙醇溶液缓慢注入到1L pH 7.4、浓度为0.05M的磷酸盐缓冲溶液中,形成乳液,然后将其转移到到圆底瓶中,在50℃水浴条件下真空旋转除去无水乙醇,形成均匀的悬浊液即为粗脂质体,形成的均匀悬浊液即为粗脂质体。将粗脂质体加入到动态高压微射流(DHPM)中,于120MPa条件下微流化处理1次,即制备得到丁香酚纳米脂质体。制得的纳米脂质体为较透明乳白色溶液。制得的丁香酚纳米脂质体的包封率为58.6%,平均粒径为58.4nm ,分布系数为0..312,在体外释放试验中,经24h释放后有77.1%的丁香酚释放出来。
实施例3
称取42.5g卵磷脂、5.7g胆固醇、10.5g吐温-80和7.1g丁香酚,完全溶解于0.31L无水乙醇中,将乙醇溶液缓慢注入 到1L pH 7.4、浓度为0.05M的磷酸盐缓冲溶液中,形成乳液,然后将其转移到到圆底瓶中,在50℃水浴条件下真空旋转除去无水乙醇,形成均匀的悬浊液即为粗脂质体,形成的均匀悬浊液即为粗脂质体。将粗脂质体加入到动态高压微射流(DHPM)中,于120MPa条件下微流化处理1次,即制备得到丁香酚纳米脂质体。制得的纳米脂质体为较透明乳白色溶液。制得的丁香酚纳米脂质体的包封率为58.9%,平均粒径为60.1nm ,分布系数为0..331,在体外释放试验中,经24h释放后有73.2%的丁香酚释放出来。
实施例4
称取55.8g卵磷脂、7.4g胆固醇、13.8g吐温-80和9.3g丁香酚,完全溶解于0.31L无水乙醇中,将乙醇溶液缓慢注入到1L pH 7.4、浓度为0.05M的磷酸盐缓冲溶液中,形成乳液,然后将其转移到到圆底瓶中,在50℃水浴条件下真空旋转除去无水乙醇,形成均匀的悬浊液即为粗脂质体,形成的均匀悬浊液即为粗脂质体。将粗脂质体加入到动态高压微射流(DHPM)中,于120MPa条件下微流化处理1次,即制备得到丁香酚纳米脂质体。制得的纳米脂质体为较透明乳白色溶液。制得的丁香酚纳米脂质体的包封率为61.2%,平均粒径为58.3nm ,分布系数为0..323,在体外释放试验中,经24h释放后有75.6%的丁香酚释放出来。
Claims (1)
1. 一种乙醇注入-动态高压微射流制备丁香酚纳米脂质体的方法,其特征是:
(1)丁香酚纳米脂质体的组分丁香酚、卵磷脂、胆固醇和吐温-80、pH=7.4浓度为 0.05M/L的磷酸盐缓冲溶液与无水乙醇的质量体积比为:在100mL pH=7.4浓度为0.05M/L的磷酸盐缓冲溶液中:丁香酚 0.38-1.16g、卵磷脂 2.32-6.99g、胆固醇 0.35-1.04g、吐温-80 0.52-1.55g、 无水乙醇 25-35mL;
(2)按上述重量百分比分别称取丁香酚、卵磷脂、胆固醇和吐温-80,在40-50℃条件下,溶于无水乙醇;
(3)将步骤(2)中所得溶液缓慢注入到pH=7.4浓度为0.05M/L的磷酸盐缓冲溶液中,在搅拌的条件下将形成乳液;
(4)将步骤(3)中所得乳液移入圆底瓶,置于真空旋转蒸发仪上,在50℃水浴条件下除去乙醇,用磷酸盐缓冲溶液定溶,得到丁香酚粗脂质体;
(5)用动态高压微射流处理步骤(4)得到的粗脂质体,压力为120MPa,处理1次,即得丁香酚纳米脂质体。
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CN105902996A (zh) * | 2016-05-27 | 2016-08-31 | 中国农业科学院农产品加工研究所 | 一种包载花生短肽的纳米脂质体及其制备方法与应用 |
CN106689241A (zh) * | 2016-11-17 | 2017-05-24 | 中国热带农业科学院农产品加工研究所 | 一种果胶包覆高良姜精油脂质体及其制备方法 |
CN113632910A (zh) * | 2021-08-06 | 2021-11-12 | 江苏省农业科学院 | 丁香酚-槐糖脂纳米乳液及其制备方法与应用 |
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