盐酸常山碱的理化性质及其脂质体包封率的测定方法考察
|
摘要
目的:研究盐酸常山碱的理化性质并考察其脂质体包封率的测定方法。方法:采用HPLC测定盐酸常山碱的含量,流动相乙睛-水-三乙胺-冰乙酸(9∶91∶0. 35∶0. 75),检测波长265 nm;通过摇瓶法测定其在不同p H范围内的油水分配系数,并考察在125℃,30 min灭菌后盐酸常山碱在不同p H磷酸盐缓冲液中的稳定性,利用硫酸铵梯度法制备盐酸常山碱脂质体,分别用葡聚糖凝胶和阳离子交换树脂制备微柱,采用离心法使游离药物和脂质体分离,测定药物含量,计算包封率。运用SPSS 20. 0软件进行t检验,比较上述2种方法测定结果的差异。结果:在pH 6~9时,盐酸常山碱的油水分配系数随着p H的升高而增大,油水分配系数处于0. 016~1. 44,盐酸常山碱灭菌后回收率在37. 16%~57. 91%,葡聚糖凝胶微柱离心法和阳离子交换树脂微柱离心法测得脂质体的包封率结果无显著性差异。结论:盐酸常山碱适合制成脂质体,但其稳定性并不理想,故在制备工艺中应严格控制实验温度。葡聚糖凝胶微柱离心法和阳离子交换树脂微柱离心法均可以较好地测定盐酸常山碱脂质体的包封率,但比较后建议选择阳离子交换树脂微柱离心法。
Objective: To study on the physical and chemical properties of dichroa alkali hydrochloride and to establish a method for the determination of entrapment efficiency of dichroa alkali hydrochloride liposomes.Method: HPLC was used to determine the content of dichroa alkali hydrochloride with mobile phase of acetonitrilewater-triethylamine-glacial acetic acid(9 ∶ 91 ∶ 0. 35 ∶ 0. 75) and detection wavelength at 265 nm. The oil-water partition coefficient of this compound in different pH range was measured by shake flask method. The stability of the dichroa alkali hydrochloride in phosphate buffer solution with different pH after sterilization at 125 ℃ for 30 min was investigated. Ammonium sulfate gradient method was used to prepare dichroa alkali hydrochloride liposomes.The microcolumn was prepared by dextran gel and cation exchange resin,respectively. Then the free drug and liposome were separated by centrifugation,the drug content was measured,and the encapsulation efficiency was calculated. The t-test was performed using SPSS 20. 0 software,the differences between these two methods were compared. Result: In the pH 6-9,the oil-water partition coefficient of dichroa alkali hydrochloride increased with increasing of pH,which was between 0. 016 and 1. 44; the recovery rate of dichroa alkali hydrochloride after sterilization was 37. 16%-57. 91%. Between the dextran gel microcolumn centrifugation and the cation exchange resin microcolumn centrifugation,there was no significant difference in the entrapment efficiency of the liposomes.Conclusion: Dichroa alkali hydrochloride is suitable for preparation of liposomes. However,its stability is not ideal,so the experimental temperature should be strictly controlled in the preparation process. Dextran gel microcolumn centrifugation and cation exchange resin microcolumn centrifugation can be used to determine the entrapment efficiency of dichroa alkali hydrochloride liposomes, and the cation exchange resin microcolumn centrifugation is suggested after comparison.
Objective: To study on the physical and chemical properties of dichroa alkali hydrochloride and to establish a method for the determination of entrapment efficiency of dichroa alkali hydrochloride liposomes.Method: HPLC was used to determine the content of dichroa alkali hydrochloride with mobile phase of acetonitrilewater-triethylamine-glacial acetic acid(9 ∶ 91 ∶ 0. 35 ∶ 0. 75) and detection wavelength at 265 nm. The oil-water partition coefficient of this compound in different pH range was measured by shake flask method. The stability of the dichroa alkali hydrochloride in phosphate buffer solution with different pH after sterilization at 125 ℃ for 30 min was investigated. Ammonium sulfate gradient method was used to prepare dichroa alkali hydrochloride liposomes.The microcolumn was prepared by dextran gel and cation exchange resin,respectively. Then the free drug and liposome were separated by centrifugation,the drug content was measured,and the encapsulation efficiency was calculated. The t-test was performed using SPSS 20. 0 software,the differences between these two methods were compared. Result: In the pH 6-9,the oil-water partition coefficient of dichroa alkali hydrochloride increased with increasing of pH,which was between 0. 016 and 1. 44; the recovery rate of dichroa alkali hydrochloride after sterilization was 37. 16%-57. 91%. Between the dextran gel microcolumn centrifugation and the cation exchange resin microcolumn centrifugation,there was no significant difference in the entrapment efficiency of the liposomes.Conclusion: Dichroa alkali hydrochloride is suitable for preparation of liposomes. However,its stability is not ideal,so the experimental temperature should be strictly controlled in the preparation process. Dextran gel microcolumn centrifugation and cation exchange resin microcolumn centrifugation can be used to determine the entrapment efficiency of dichroa alkali hydrochloride liposomes, and the cation exchange resin microcolumn centrifugation is suggested after comparison.
引文
[1] World Health Organization. Word malaria report 2017[M]. Geneva:World Health Organization,2017.
[2] Imwong M,Suwannasin K,Kunasol C,et al. The spread of artemisinin-resistant Plasmodium falciparum in the Greater Mekong subregion:a molecular epidemiology observational study[J]. Lancet Infect Dis,2017,17(5):491-497.
[3] Grigg M J,William T,Menon J,et al. Efficacy of artesunate-mefloquine for chloroquine-resistant plasmodium vivax malaria in Malaysia:an open-label,randomized,controlled trial[J]. Clin Infect Dis,2016,62(11):1403-1411.
[4] Mishra M,Mishra V K,Kashaw V,et al. Comprehensive review on various strategies for antimalarial drug discovery[J]. Eur J Med Chem,2016,125:1300-1320.
[5]叶祖光,李春,李思迪,等.常山碱盐酸盐及其制备方法和医药应用:中国,CN105535045A[P]. 2016-05-04.
[6]李思迪,李春,叶祖光,等.常山碱盐急性毒性及其与青蒿素类药物联合用药增效减毒作用[J].中国药理学与毒理学杂志,2016,30(8):808-814.
[7] Bozzuto G,Molinari A. Liposomes as nanomedical devices[J]. Int J Nanomedicine,2015,10:975-999.
[8] Aliu H,Rask C,Brimnes J,et al. Enhanced efficacy of sublingual immunotherapy by liposome-mediated delivery of allergen[J]. Int J Nanomedicine,2017,12:8377-8388.
[9] Allen T M,Cullis P R. Liposomal drug delivery systems:from concept to clinical applications[J]. Adv Drug Deliv Rev,2013,65(1):36-48.
[10]陈斯泽,汪森明,张积仁.盐酸洛拉曲克长循环脂质体制备及其抑瘤特性研究[J].南方医科大学学报,2008,28(3):403-405.
[11]张慧慧,聂淑芳,李想,等.马来酸噻吗洛尔眼用脂质体的研制及其离体角膜透过试验[J].中国新药杂志,2009,18(16):1556-1559.
[12]黄园,段逸松,于波涛,等.硫酸铵梯度法制备米托蒽醌长循环脂质体[J].中国药学杂志,2002,37(12):917-919.
[13]胡鹏翼,郑琴,杨明,等.主动载药法制备槐定碱脂质体[J].中国实验方剂学杂志,2011,17(13):5-8.
[14]余昕,陈斯玮,欧丽兰,等.芍药总苷的理化性质及其脂质体包封率的测定方法[J].中成药,2016,38(6):1260-1264.
[15]康传哲,张明珠,陈岩,等. 10-甲氧基喜树碱平衡溶解度和表观油水分配系数的测定[J].中国实验方剂学杂志,2014,20(9):133-136.
[16]吴玉强,王志萍,熊万娜,等.秋水仙碱油水分配系数和平衡溶解度的测定[J].中国实验方剂学杂志,2016,22(5):64-66.
[17]张琪,黄娟,薛鸿娇,等.乙酰天麻素平衡溶解度、油水分配系数及稳定性的测定[J].中国医院药学杂志,2017,37(9):819-821.
[18]郝朵,谷福根. p H值对富马酸比索洛尔化学稳定性及油水分配系数的影响[J].中国医院药学杂志,2013,33(3):208-211.
[19]李文静,杨志强,王杏林.两种硫酸长春新碱脂质体包封率测定方法的比较[J].中国实验方剂学杂志,2012,18(14):71-75.
[20]张玲,黄微崴,邓意辉,等.重酒石酸长春瑞滨脂质体包封率测定方法比较[J].沈阳药科大学学报,2010,27(2):105-109.
[21]周金幸,顾开龙,胡海洋,等. Box-Behnken效应面法优化制备主动载药盐酸小檗碱脂质体[J].沈阳药科大学学报,2014,31(11):856-861.
[22]郑杭生,张亚军,徐真,等.全缘千里光碱脂质体包封率测定方法的研究[J].中国药学杂志,2007,42(14):1075-1078.
[23]陈孝储,郭一沙,宋菲,等.微柱离心结合胆固醇浓度变化测定水溶性药物脂质体包封率[J].沈阳药科大学学报,2014,31(11):851-855.
[24]聂华,赵莹,叶小玲,等.葡萄糖修饰脑靶向紫杉醇脂质体包封率测定方法的比较[J].中国实验方剂学杂志,2016,22(23):15-19.
[25]赵妍,于彬,邓意辉,等.主动载药法制备硫酸长春新碱脂质体及其包封率的测定[J].中国药学杂志,2005,39(20):1559-1562.
[26] Tamaru S,Igura N,Shimoda M. Effectiveness of water-air and octanol-air partition coefficients to predict lipophilic flavor release behavior from O/W emulsions[J]. Food Chem,2018,239:712-717.
[27] BAO Q Y,LIU A Y,MA Y,et al. The effect of oil-water partition coefficient on the distribution and cellular uptake of liposome-encapsulated gold nanoparticles[J].Colloid Surface B,2016,146:475-481.
[28] Yamamoto E,Miyazaki S,Aoyama C,et al. A simple and rapid measurement method of encapsulation efficiency of doxorubicin loaded liposomes by direct injection of the liposomal suspension to liquid chromatography[J]. Int J Pharm,2018,536(1):21-28.
[29]陈召红,刘皈阳,魏亚超.脂质体包封率测定方法研究进展[J].解放军药学学报,2011,27(1):79-82.
[2] Imwong M,Suwannasin K,Kunasol C,et al. The spread of artemisinin-resistant Plasmodium falciparum in the Greater Mekong subregion:a molecular epidemiology observational study[J]. Lancet Infect Dis,2017,17(5):491-497.
[3] Grigg M J,William T,Menon J,et al. Efficacy of artesunate-mefloquine for chloroquine-resistant plasmodium vivax malaria in Malaysia:an open-label,randomized,controlled trial[J]. Clin Infect Dis,2016,62(11):1403-1411.
[4] Mishra M,Mishra V K,Kashaw V,et al. Comprehensive review on various strategies for antimalarial drug discovery[J]. Eur J Med Chem,2016,125:1300-1320.
[5]叶祖光,李春,李思迪,等.常山碱盐酸盐及其制备方法和医药应用:中国,CN105535045A[P]. 2016-05-04.
[6]李思迪,李春,叶祖光,等.常山碱盐急性毒性及其与青蒿素类药物联合用药增效减毒作用[J].中国药理学与毒理学杂志,2016,30(8):808-814.
[7] Bozzuto G,Molinari A. Liposomes as nanomedical devices[J]. Int J Nanomedicine,2015,10:975-999.
[8] Aliu H,Rask C,Brimnes J,et al. Enhanced efficacy of sublingual immunotherapy by liposome-mediated delivery of allergen[J]. Int J Nanomedicine,2017,12:8377-8388.
[9] Allen T M,Cullis P R. Liposomal drug delivery systems:from concept to clinical applications[J]. Adv Drug Deliv Rev,2013,65(1):36-48.
[10]陈斯泽,汪森明,张积仁.盐酸洛拉曲克长循环脂质体制备及其抑瘤特性研究[J].南方医科大学学报,2008,28(3):403-405.
[11]张慧慧,聂淑芳,李想,等.马来酸噻吗洛尔眼用脂质体的研制及其离体角膜透过试验[J].中国新药杂志,2009,18(16):1556-1559.
[12]黄园,段逸松,于波涛,等.硫酸铵梯度法制备米托蒽醌长循环脂质体[J].中国药学杂志,2002,37(12):917-919.
[13]胡鹏翼,郑琴,杨明,等.主动载药法制备槐定碱脂质体[J].中国实验方剂学杂志,2011,17(13):5-8.
[14]余昕,陈斯玮,欧丽兰,等.芍药总苷的理化性质及其脂质体包封率的测定方法[J].中成药,2016,38(6):1260-1264.
[15]康传哲,张明珠,陈岩,等. 10-甲氧基喜树碱平衡溶解度和表观油水分配系数的测定[J].中国实验方剂学杂志,2014,20(9):133-136.
[16]吴玉强,王志萍,熊万娜,等.秋水仙碱油水分配系数和平衡溶解度的测定[J].中国实验方剂学杂志,2016,22(5):64-66.
[17]张琪,黄娟,薛鸿娇,等.乙酰天麻素平衡溶解度、油水分配系数及稳定性的测定[J].中国医院药学杂志,2017,37(9):819-821.
[18]郝朵,谷福根. p H值对富马酸比索洛尔化学稳定性及油水分配系数的影响[J].中国医院药学杂志,2013,33(3):208-211.
[19]李文静,杨志强,王杏林.两种硫酸长春新碱脂质体包封率测定方法的比较[J].中国实验方剂学杂志,2012,18(14):71-75.
[20]张玲,黄微崴,邓意辉,等.重酒石酸长春瑞滨脂质体包封率测定方法比较[J].沈阳药科大学学报,2010,27(2):105-109.
[21]周金幸,顾开龙,胡海洋,等. Box-Behnken效应面法优化制备主动载药盐酸小檗碱脂质体[J].沈阳药科大学学报,2014,31(11):856-861.
[22]郑杭生,张亚军,徐真,等.全缘千里光碱脂质体包封率测定方法的研究[J].中国药学杂志,2007,42(14):1075-1078.
[23]陈孝储,郭一沙,宋菲,等.微柱离心结合胆固醇浓度变化测定水溶性药物脂质体包封率[J].沈阳药科大学学报,2014,31(11):851-855.
[24]聂华,赵莹,叶小玲,等.葡萄糖修饰脑靶向紫杉醇脂质体包封率测定方法的比较[J].中国实验方剂学杂志,2016,22(23):15-19.
[25]赵妍,于彬,邓意辉,等.主动载药法制备硫酸长春新碱脂质体及其包封率的测定[J].中国药学杂志,2005,39(20):1559-1562.
[26] Tamaru S,Igura N,Shimoda M. Effectiveness of water-air and octanol-air partition coefficients to predict lipophilic flavor release behavior from O/W emulsions[J]. Food Chem,2018,239:712-717.
[27] BAO Q Y,LIU A Y,MA Y,et al. The effect of oil-water partition coefficient on the distribution and cellular uptake of liposome-encapsulated gold nanoparticles[J].Colloid Surface B,2016,146:475-481.
[28] Yamamoto E,Miyazaki S,Aoyama C,et al. A simple and rapid measurement method of encapsulation efficiency of doxorubicin loaded liposomes by direct injection of the liposomal suspension to liquid chromatography[J]. Int J Pharm,2018,536(1):21-28.
[29]陈召红,刘皈阳,魏亚超.脂质体包封率测定方法研究进展[J].解放军药学学报,2011,27(1):79-82.
© 2004-2018 中国地质图书馆版权所有 京ICP备05064691号 京公网安备11010802017129号 地址：北京市海淀区学院路29号 邮编：100083 电话：办公室：(+86 10)66554848；文献借阅、咨询服务、科技查新：66554700 |