油包水微乳的制备及特性研究
|
摘要
亲水性药物口服后容易被胃蛋白酶严重破坏,而且分子体积较大难以被肠道粘膜吸收,限制了亲水性药物的生物利用度。油包水(W/O型)微乳可以增强亲水性药物在肠道粘膜上的通透性,提高了药物口服的生物利用度。鉴于微乳所用表面活性剂大多在食品中有用量限制,本研究旨在筛选W/O型微乳的最优配方,寻求微乳的高载药量和低表面活性剂含量,探究了以span80-tween80,PEG-7氢化蓖麻油和Labrasol-plurol diisostearique为表面活性剂制备微乳时Km(表面活性剂和助表面活性剂的比值)值和油相链长对微乳相区面积的影响。考察了微乳的结构类型,水相中NaCl浓度和温度对微乳相区面积的影响以及水相中包埋不同分子量的氨基酸,多肽和蛋白对微乳相区面积影响的规律性。同时考察了胰岛素微乳的降血糖效果,5-氟尿嘧啶(5-Fu)微乳的缓释效果以及微乳的储存稳定性。
研究发现,以span80-tween80 (HLB=6)和PEG-7氢化蓖麻油为表面活性剂,以乙醇为助表面活性剂时最适合于制备W/O型微乳并用于亲水性药物载体,两者形成W/O型微乳最佳Km值分别是4:1和3:1,制备W/O型微乳的最优配方为:肉豆蔻酸异丙酯含量61.6%,混合表面活性剂(表面活性剂和助表面活性剂混合物)含量26.4%,水相含量12%。通过电导率测定,流变性质测定以及染色法鉴定证明三类表面活性剂形成的微乳均为W/O型微乳。微乳相区面积随着温度升高略微增大、随着NaCl浓度增加相区面积有所减小。另外,水相中包埋不同分子量的氨基酸,多肽和蛋白质其微乳相区面积呈现规律性变化,氨基酸主要通过影响微乳界面膜形成来影响微乳相区面积,多肽和蛋白质主要通过影响微乳水相的黏度来影响微乳相区面积。包埋氨基酸微乳相区面积随着氨基酸浓度增大基本呈现先减小后增加趋势,而包埋多肽和蛋白质微乳相区面积随着多肽和蛋白质浓度增大呈现逐渐减小趋势。通过不同碳链油相形成微乳相区面积的比较发现中等碳链长度的肉豆蔻酸异丙酯最适合作为制备微乳的油相。微乳粒径在10~30nm范围,多分散指数(PDI)在0.08~0.4范围内,具有良好的均一性和稳定性。
以最优配方制备W/O型微乳载入胰岛素,通过小鼠的口服胰岛素微乳血糖测定结果表明,口服胰岛素微乳具有显著的降血糖效果,span80-tween80 (HLB=6)和PEG-7氢化蓖麻油两种胰岛素微乳制剂在小鼠口服葡糖糖的代谢过程中,第60min时相对于口服空白样品血糖相对值(相对于初始血糖值)分别降低183%和151%。两种胰岛素微乳的水相中分别包埋质量分数为1%和3%的大豆蛋白酶抑制剂和牛磺胆酸盐可以进一步提高口服胰岛素微乳的降血糖效果。并且随着包埋物中蛋白酶抑制剂和牛磺胆酸盐浓度增加,胰岛素微乳的降血糖效果也有所提高。其中,在第60min时,口服含胰蛋白酶抑制剂的span80-tween80 (HLB=6)胰岛素微乳相对于空白样品血糖相对值降低231%。
以最优配方制备W/O型微乳载入5-氟尿嘧啶(5-Fu),通过5-Fu微乳在模拟胃肠pH环境中的体外释放实验结果表明,span80-tween80和PEG-7氢化蓖麻油微乳对水相中的5-Fu具有明显的缓释作用。在span80-tween80 (HLB=6)微乳中,5-Fu在模拟胃液和肠液中20h时累积释放率分别是60.41%和87.04%;在PEG-7氢化蓖麻油微乳中,5-Fu在模拟胃液和肠液中20h时累积释放率分别是77.83%和62.46%,而5-Fu水溶液,在20h累积释放率分别达到90%和99%。另外,水相中添加一定浓度的NaCl可以提高微乳的缓释效果。其中,水相中添加0.1mol/L NaCl的span80-tween80 (HLB=6)微乳在模拟肠液中相对于空白微乳20h时累积释放率降低了13.76%。
微乳稳定性实验表明,span80-tween80 (HLB=6)和PEG-7氢化蓖麻油制备的微乳具有较强的稳定性,储藏过程中微乳粒径以及PDI均没有显著变化。同时在0℃和25℃时储藏两个月的微乳并未出现分层和浑浊现象,在6000r/min条件下离心也未出现分层现象。
When administered orally, Hydrophilic drug undergoes severe enzymatic degragation by gastric proteases and its high molecular weight renders it impermeable across gastric mucosa, limiting its overall bioavailability. Water in oil (W/O) microemulsion has a significant potential to increase permeability of hydrophilic drug across the intestinal mucosa and thus enhance their oral bioavailability. Using surfactants to stabilize microemulsions is associated with dosage limited in the food Field, this study aimed to screening optimal formula of W/O microemulsion, ensuring high drug loading and low surfactants content. The preparation of microemulsions using span80-tween80, PEG-7 hydrogenated castor oil and Labrasol-plurol diisostearique as surfactants and the effects of Km (the ratio of surfactant to cosurfactant) as well as the chain length of oil phase on microemulsion phase area were investigated. The structures of microemulsion, the effects of NaCl concentrations and temperatures on the microemulsions phase area were also examined. Also amino acids, peptides and proteins with different molecular weights and their associated effects on the phase behavior were evaluated. Hypoglycemic effects of insulin microemulsion, sustained release ability of 5-Fu microemulsion and the storage stability of the microemulsion were investigated.
The results indicated that surfactants span80-tween80 and PEG-7 hydrogenated castor oil were suitable as hydrophilic drug carriers for preparation of W/O microemulsion and the optimum Km were 4:1 and 3:1, respectively. The optimal formula for preparation of W/O microemulsion was 61.6%, 26.3% and 12% of isopropyl myristate, mixed surfactants, and aqueous phase respectively. The conductivity, rheological properties and staining method showed that the three types of surfactants used all formed W/O microemulsions. Microemulsion phase area increased slightly as temperature increased and lowered when concentration of NaCl solution was elevated. The addition of amino acids, peptides and proteins with different molecular weight to the aqueous phase resulted in a regular change of phase area of microemulsion. While amino acids mainly through their effect on interface membrane formation influenced the microemulison phase area, peptide and protein did so through the viscosity of the aqueous phase of microemulsion phase area. The microemulsion phase area firstly showed both decreasing and then increasing trend when amino acids concentration increased in the aqueous phase. However, when peptide and protein concentration increased in the aqueous phase, the microemulsion phase area decreased. The comparison of the phase areas of microemulsions with different carbon chain showed that isopropyl myristate with medium carbon chain length was the most suitable oil phase for preparation of microemulsion. The particle diameter ranged from 10 to 30 nm and the Polydispersity index ranged from 0.08 to 0.40 which indicated that microemulsion had good uniformity and stability.
The determination of blood glucose level of initial blood glucose level (BGL) after intake of insulin by experimental mice indicated that oral insulin exerted a significant hypoglycemic effect and when compared with those of control groups, the BGL of mice treated with the Span80-tween80 and PEG-7 hydrogenated castor oil microemulsions containing insulin was lowered by 183% and 151% respectively within 60 minutes. Hypoglycemic effect of oral insulin was enhanced with the entrapment of 1% (w/w) trypsin inhibitor and 3% (w/w) sodium taurocholate in aqueous phase. In the case of Span80-tween80 (HLB=6) microemulsion system, entrapment of 3% trypsin inhibitor in aqueous phase reduced the BGL of mice treated with insulin by 231% within 60 minutes.
The sustained release test of 5-Fu mieroemulsion in the pH of simulated gastrointestinal tract environment indicated that Span80-tween80 and PEG-7 hydrogenated castor oil systems had a significant sustained release effect on 5-Fu. Span80-tween80 and PEG-7 hydrogenated castor oil systems gave total release rates of 59.6% and 77.83% in a simulated gastric juice and yielded 87% and 63.40% in a simulated intestinal juice after 20 hours, respectively. At the same time, the 5-Fu aqueous solution had a total release ratio of 90% after 20 hours. Entrapment of 1% NaCl in aqueous phase of Span80-tween80 system reduced the total release rate of 5-Fu in a simulated intestinal juice by 13.76% within 20 hours.
The stability test indicated that microemulsions prepared by Span80-tween80 and PEG-7 hydrogenated castor oil had good stabilities and during storage process the particle diameter and polydispersity index showed no significant change. Multiple-layer and turbidity didn’t appear in the microemulsions during two month storage at 0°C and 25°C or by centrifugation at 6000 rpm.
研究发现,以span80-tween80 (HLB=6)和PEG-7氢化蓖麻油为表面活性剂,以乙醇为助表面活性剂时最适合于制备W/O型微乳并用于亲水性药物载体,两者形成W/O型微乳最佳Km值分别是4:1和3:1,制备W/O型微乳的最优配方为:肉豆蔻酸异丙酯含量61.6%,混合表面活性剂(表面活性剂和助表面活性剂混合物)含量26.4%,水相含量12%。通过电导率测定,流变性质测定以及染色法鉴定证明三类表面活性剂形成的微乳均为W/O型微乳。微乳相区面积随着温度升高略微增大、随着NaCl浓度增加相区面积有所减小。另外,水相中包埋不同分子量的氨基酸,多肽和蛋白质其微乳相区面积呈现规律性变化,氨基酸主要通过影响微乳界面膜形成来影响微乳相区面积,多肽和蛋白质主要通过影响微乳水相的黏度来影响微乳相区面积。包埋氨基酸微乳相区面积随着氨基酸浓度增大基本呈现先减小后增加趋势,而包埋多肽和蛋白质微乳相区面积随着多肽和蛋白质浓度增大呈现逐渐减小趋势。通过不同碳链油相形成微乳相区面积的比较发现中等碳链长度的肉豆蔻酸异丙酯最适合作为制备微乳的油相。微乳粒径在10~30nm范围,多分散指数(PDI)在0.08~0.4范围内,具有良好的均一性和稳定性。
以最优配方制备W/O型微乳载入胰岛素,通过小鼠的口服胰岛素微乳血糖测定结果表明,口服胰岛素微乳具有显著的降血糖效果,span80-tween80 (HLB=6)和PEG-7氢化蓖麻油两种胰岛素微乳制剂在小鼠口服葡糖糖的代谢过程中,第60min时相对于口服空白样品血糖相对值(相对于初始血糖值)分别降低183%和151%。两种胰岛素微乳的水相中分别包埋质量分数为1%和3%的大豆蛋白酶抑制剂和牛磺胆酸盐可以进一步提高口服胰岛素微乳的降血糖效果。并且随着包埋物中蛋白酶抑制剂和牛磺胆酸盐浓度增加,胰岛素微乳的降血糖效果也有所提高。其中,在第60min时,口服含胰蛋白酶抑制剂的span80-tween80 (HLB=6)胰岛素微乳相对于空白样品血糖相对值降低231%。
以最优配方制备W/O型微乳载入5-氟尿嘧啶(5-Fu),通过5-Fu微乳在模拟胃肠pH环境中的体外释放实验结果表明,span80-tween80和PEG-7氢化蓖麻油微乳对水相中的5-Fu具有明显的缓释作用。在span80-tween80 (HLB=6)微乳中,5-Fu在模拟胃液和肠液中20h时累积释放率分别是60.41%和87.04%;在PEG-7氢化蓖麻油微乳中,5-Fu在模拟胃液和肠液中20h时累积释放率分别是77.83%和62.46%,而5-Fu水溶液,在20h累积释放率分别达到90%和99%。另外,水相中添加一定浓度的NaCl可以提高微乳的缓释效果。其中,水相中添加0.1mol/L NaCl的span80-tween80 (HLB=6)微乳在模拟肠液中相对于空白微乳20h时累积释放率降低了13.76%。
微乳稳定性实验表明,span80-tween80 (HLB=6)和PEG-7氢化蓖麻油制备的微乳具有较强的稳定性,储藏过程中微乳粒径以及PDI均没有显著变化。同时在0℃和25℃时储藏两个月的微乳并未出现分层和浑浊现象,在6000r/min条件下离心也未出现分层现象。
When administered orally, Hydrophilic drug undergoes severe enzymatic degragation by gastric proteases and its high molecular weight renders it impermeable across gastric mucosa, limiting its overall bioavailability. Water in oil (W/O) microemulsion has a significant potential to increase permeability of hydrophilic drug across the intestinal mucosa and thus enhance their oral bioavailability. Using surfactants to stabilize microemulsions is associated with dosage limited in the food Field, this study aimed to screening optimal formula of W/O microemulsion, ensuring high drug loading and low surfactants content. The preparation of microemulsions using span80-tween80, PEG-7 hydrogenated castor oil and Labrasol-plurol diisostearique as surfactants and the effects of Km (the ratio of surfactant to cosurfactant) as well as the chain length of oil phase on microemulsion phase area were investigated. The structures of microemulsion, the effects of NaCl concentrations and temperatures on the microemulsions phase area were also examined. Also amino acids, peptides and proteins with different molecular weights and their associated effects on the phase behavior were evaluated. Hypoglycemic effects of insulin microemulsion, sustained release ability of 5-Fu microemulsion and the storage stability of the microemulsion were investigated.
The results indicated that surfactants span80-tween80 and PEG-7 hydrogenated castor oil were suitable as hydrophilic drug carriers for preparation of W/O microemulsion and the optimum Km were 4:1 and 3:1, respectively. The optimal formula for preparation of W/O microemulsion was 61.6%, 26.3% and 12% of isopropyl myristate, mixed surfactants, and aqueous phase respectively. The conductivity, rheological properties and staining method showed that the three types of surfactants used all formed W/O microemulsions. Microemulsion phase area increased slightly as temperature increased and lowered when concentration of NaCl solution was elevated. The addition of amino acids, peptides and proteins with different molecular weight to the aqueous phase resulted in a regular change of phase area of microemulsion. While amino acids mainly through their effect on interface membrane formation influenced the microemulison phase area, peptide and protein did so through the viscosity of the aqueous phase of microemulsion phase area. The microemulsion phase area firstly showed both decreasing and then increasing trend when amino acids concentration increased in the aqueous phase. However, when peptide and protein concentration increased in the aqueous phase, the microemulsion phase area decreased. The comparison of the phase areas of microemulsions with different carbon chain showed that isopropyl myristate with medium carbon chain length was the most suitable oil phase for preparation of microemulsion. The particle diameter ranged from 10 to 30 nm and the Polydispersity index ranged from 0.08 to 0.40 which indicated that microemulsion had good uniformity and stability.
The determination of blood glucose level of initial blood glucose level (BGL) after intake of insulin by experimental mice indicated that oral insulin exerted a significant hypoglycemic effect and when compared with those of control groups, the BGL of mice treated with the Span80-tween80 and PEG-7 hydrogenated castor oil microemulsions containing insulin was lowered by 183% and 151% respectively within 60 minutes. Hypoglycemic effect of oral insulin was enhanced with the entrapment of 1% (w/w) trypsin inhibitor and 3% (w/w) sodium taurocholate in aqueous phase. In the case of Span80-tween80 (HLB=6) microemulsion system, entrapment of 3% trypsin inhibitor in aqueous phase reduced the BGL of mice treated with insulin by 231% within 60 minutes.
The sustained release test of 5-Fu mieroemulsion in the pH of simulated gastrointestinal tract environment indicated that Span80-tween80 and PEG-7 hydrogenated castor oil systems had a significant sustained release effect on 5-Fu. Span80-tween80 and PEG-7 hydrogenated castor oil systems gave total release rates of 59.6% and 77.83% in a simulated gastric juice and yielded 87% and 63.40% in a simulated intestinal juice after 20 hours, respectively. At the same time, the 5-Fu aqueous solution had a total release ratio of 90% after 20 hours. Entrapment of 1% NaCl in aqueous phase of Span80-tween80 system reduced the total release rate of 5-Fu in a simulated intestinal juice by 13.76% within 20 hours.
The stability test indicated that microemulsions prepared by Span80-tween80 and PEG-7 hydrogenated castor oil had good stabilities and during storage process the particle diameter and polydispersity index showed no significant change. Multiple-layer and turbidity didn’t appear in the microemulsions during two month storage at 0°C and 25°C or by centrifugation at 6000 rpm.
引文
1. Jonsson B, lindman B, Holmberg K, Kronberg B. Surfactants and Polymers in Aqueous Solution[J].John Wiley&Sons,Chichester,1998.
2. Hoar T P, Schulman J H. Transparent water-in-oil dispersions the oleopathic hydro-micelle [J]. Nature, 1943, 152: 102-03:21.
3. Jayne M L and Gareth D R. Microemulsion-based media as novel drug delivery systems[J]. Advanced Drug Delivery Reviews, 45(2000): 89-121.
4. Cilek, Celebi N, Tirnaksiz F, et al.. Tay A. A lecithin-based microemulsion of rh-insulin with aprotinin for oral administration: investigation of hypoglycemic effects in non-diabetic and STZ-induced diabetic rats[J]. International Journal of Pharmaceutics, 298(2005): 176-185.
5. Patel G, Misra A. Oral Delivery of Proteins and Peptides: Concepts and Applications[J]. Challenges in Delivery of Therapeutic Genomics and Proteomics, 2011: 481-529.
6. Tarver T. Food nanotechnology[J].Food Technology,2006,60(11):22-26
7. Chen H, Weiss J, Shahidi F, et al.. Nanotechnology in nutraceuticals and functional foods[J].Food Technology, 2006, 60(3):30-32.
8. Panayiotis P C, Scalart J P. Formulation and physical characterization of water in oil microemulsions containing long versus medium chain glycerides[J]. International Journal of Pharmaceutics, 158(1997):57-68
9. Guo-Rong Duan, Xu-Jie Yang, Guo-Hong Huang, Lu-De Lu. Water/span80/Triton X-100/n-hexyl alcohol/n-octane microemulsion system and the study of its application for preparing nanosized zirconia[J]. Materials Letters, 60(2006) 1582-1587.
10. Yiming Guo, Jingzhe Zhao, et al.. Preparation and characterization of monoclinic sulfur nanoparticles by water-in-oil microemulsions technique[J]. Powder Technology, 162 (2006):83-86.
11. Garti N, Aserin A, Fanun M. Non-ionic sucrose esters microemulsions for food applications. Part 1. Water solubilization[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 164(2000): 27-38.
12. Glatter O, Orthaber D, Stradner A, et al.. Sugar-Ester Nonionic Microemulsion: Structural Characterization[J]. Journal of Colloid and Interface Science, 241(2001):215-225.
13. Kreilgaard M, Pedersen E J, Jaroszewski J W. NMR characterisation and transdermal drug delivery potential of microemulsion systems[J]. Journal of Controlled Release,69(2000):421-433.
14. Sintova A C, Levy H V, Botner S, et al.. Systemic delivery of insulin via the nasal route using a new microemulsion system: In vitro and in vivo studies[J]. Journal of Controlled Release, 148(2010):168-176.
15. Graf A, Ablingerb E, Peters S, et al.. Microemulsions containing lecithin and sugar based surfactants Nanoparticle templates for delivery of proteins and peptides[J]. International Journal of Pharmaceutics, 350 (2008): 351-360.
16. Li-Ching Peng, Chi-Hsien Liub, Chang-Chin Kwana, Keh-Feng Huang. Optimization of water-in-oil nanoemulsions by mixed surfactants[J]. Colloids and Surfaces A: Physicochem. Eng. Aspects, 370 (2010):136-142.
17.路莹,吉小欣,高申,刘毅清.利多卡因-卵磷脂微乳处方的初步研究[J].药学实践杂志, 2004 22(3):141-143.
18. Alany R G, Rades T, Kustrin S A, et al.. Effects of alcohols and diols on the phase behaviour of quaternary systems[J]. International Journal of Pharmaceutics, 196(2000): 141-145.
19. Warisnoicharoen W, Lansley A B, Lawrence M J. Nonionic oil-in-water microemulsions: the effect of oil type on phase behaviour[J]. Int J Pharm, 2000(198):7-27.
20. Attwood D. Microemulsions [J]: Colloidal Drug Delivery Systems, Dekker, New 3171.
21. Constantinides P P, Seang H Y, et al.. Particle size determination of phase-inverted water-in-oil microemulsions under different dilution and storage conditions[J]. International Journal of Pharmaceutics, 115(1995):225-234.
22. Lutz Schlicht, Jan-Hendrik Spilgies, Georg Ilgenfritz. Dynamics of Structure Changes in Water-in-Oil Microemulsions: Electric Birefringence and Electric Light Scattering in Percolating Systems[J]. Journal of Molecular Liquids, 72(1997):295-314.
23. Fernandez J C, Bisceglia M, Acosta E, et al.. On the birefringence of water-alkane-AOT microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 157(1999):35-46.
24. Acosta E, Kurlat D H, Bisceglia M. Induced electric birefringence and viscosity studies in microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 106(1996):11-21.
25. Eicke H F, Hilfiker R. Connectivity-controlled charge transport in water-in-oil microemulsions[J]. Chemical Physics Letters, 1986,125(3):295-298.
26. Rozieres J, Marcia A. Middleton and Robert S, et al.. Schechter.Theoretical model for the complex permittivity of water-in-oil microemulsions with ionic surfactants[J]. Journal of Colloid and Interface Science, 1988 124(3):407-415.
27. Senatra D. Dielectric analysis and Differential Scanning Calorimetry of water-in-oil microemulsions[J]. Advances in Colloid and Interface Science, 123-126 (2006):415-424.
28. Senatra D, Ziparo C, Cecilia M. C. Gambi1, L. Lanzi, ENERGY BALANCE IN DENSE MICROEMULSIONS[J]. Journal of Thermal Analysis and Calorimetry, 2008 92 (2): 535-541.
29. Senatral D, Zhoul Z, Pieraccini L, et al.. A study of the properties of water-in-oil microemulsions in the subzero temperature range by differential scanning calorimetry[J]. Progr Colloid & Polymer Sci, 73(1987):66-75.
30. Garti N, Aserin A, Tiunova I, Fanun M. A DSC study of water behavior in water-in-oil microemulsions stabilized by sucrose esters and butanol[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 170(2000) 1-18.
31. Senatra D. Procedures for DSC analysis of percolative microemulsions[J]. Thermochimica Acta, 345 (2000) 39-46.
32. Campo L, Yaghmur A, et al.. Five-component food-grade microemulsions: structural characterization by SANS[J]. Journal of Colloid and Interface Science, 274 (2004):251–267.
33. Azouza B, Oberb R, Nakache E, Williams C E. A small angle X-ray scattering investigation of the structure of a ternary water-in-oil microemulsion[J]. Colloids and Surfaces, 1992,69(2-3): 87-97.
34. Behera G B, Mishra B K, Behera P K, Panda M. Fluorescent probes for structural and distance effect studies in micelles, reversed micelles and microemulsions[J]. Advances in Colloid and Interface Science, 82 (1999):1-42.
35. Nornoo A O, Zheng H, Lopes L B, et al.. Oral microemulsions of paclitaxel: In situ and pharmacokinetic studies[J]. European Journal of Pharmaceutics and Biopharmaceutics, 71(2009):310–317.
36. Walderhaug H. Microstructures in Aqueous Solutions of a Polyoxyethylene Trisiloxane Surfactant and a Cosurfactant Studied by SANS and NMR Self-Diffusion[J]. Langmuir, 2008(24):10637-10645.
37. Mehta S K, Kaur G, Bhasin K K, et al.. Tween-Embedded Microemulsions Physicochemical and Spectroscopic Analysis for Antitubercular Drugs[J]. AAPS PharmSciTech, 2010(11):143-153.
38. Fanun M. A study of the properties of mixed nonionic surfactants microemulsions by NMR, SAXS, viscosity and conductivity[J]. Journal of Molecular Liquids, 142(2008): 103-110.
39. Fanun M, Wail S A. Electrical conductivity and self diffusion NMR studies of the system:Water/sucrose laurate/ethoxylated monodiglyceride/isopropylmyristate[J]. Colloids and Surfaces A: Physicochem. Eng. Aspects, 277 (2006):83-89.
40. Zuliang Lai, Peiyi Wu. Investigation on the conformations of AOT in water-in-oil microemulsions using 2D-ATR-FTIR correlation spectroscopy[J]. Journal of Molecular Structure, 883-884 (2008):236-241.
41.梁文平,乳状液科学与技术基础[M],北京:科学出版社,2001,211-245.
42. Guptaa R R, Swantrant K. Jainb, Manoj Varshney, et al.. AOT water-in-oil microemulsions as a penetration enhancer in transdermal drug delivery of 5-?uorouracil[J]. Colloids and Surfaces B: Biointerfaces, 41 (2005):25-32.
43. Dracha M, Micha?eka J N, Sienkiewicz A. Antioxidative properties of vitamins C and E in micellar systems and in microemulsions[OL]. Colloids and Surfaces A: Physicochem. Eng, 2010.
44. Avramiotis S, Papadimitriou V, Cazianis C T, A. Xenakis. EPR studies of proteolytic enzymes in microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 144(1998):295-304.
45. Sharma G, Wilson K, Walle C F, et al.. Microemulsions for oral delivery of insulin: Design, development and evaluation in streptozotocin induced diabetic rats[J]. European Journal of Pharmaceutics and Biopharmaceutics, 76 (2010): 159-169.
46. Graf A, Rades T, Hook S M. Oral insulin delivery using nanoparticles based on microemulsions with different structure-types: Optimisation and in vivo evaluation[J]. european journal of pharmaceutical sciences, 37 (2009): 53-61.
47. Actis G C, Lagge M, Rizzetto M, et al.. Long-term efficacy of oral microemulsion cyclosporin for refractory ulcerative colitis[J].Minerva lVled,2004,95(1): 65-70.
48. Peira E, Scolari P, Gasco M R. Transdermal permeation of apomorphine through hairless mouse skin from microemulsions[J]. International Journal of Pharmaceutics, 226 (2001): 47-51.
49.王建磊,王正武,俞惠新.胰岛素W/O型微乳液的制备及体外释药性能[J].精细化学2006,23(9): 867-872.
50. Mao-Bo Cheng, Jian-Cheng Wang, Yu-Hua Li, Characterization of water-in-oil microemulsion for oral delivery of earthworm fibrinolytic enzyme[J]. Journal of Controlled Release, 129, 2008: 41-48.
51.韩旻,傅韶,方晓玲.三七总皂苷油包水微乳的处方筛选及体内外评价[J].药学学报2007,42(7): 780-786.
52.孟博宇,平其能,熊净,高琴.胰岛素微乳大鼠肠道给药降糖作用的研究[J].中国药科大学学报.2006,37(4): 304-307.
53. Lyons K C, Charman W N, Miller M, et al. Factors limiting the oral bioavailability of N-acetylglucosaminyl-N-acetylmuramyl dipeptide (GMDP) and enhancement of absorption in rats by delivery in a water-in-oil microemulsion[J]. International Journal of Pharmaceutics, 199 (2000): 17-28.
54. Damge C, Maincent P. Oral delivery of insulin associated to polymeric nanoparticles in diabetic mice[J]. Journal of Controlled Release.117. 2007:163–170.
55. Zelihagül Degim, Nilayünal. The effect of various liposome formulations on insulin[J]. Life Sciences. 75(23),2004: 2819-2827.
56. Cournarie F, Savelli M P. Insulin-loaded W/O/W multiple emulsions [J]. European Journal of Pharmaceutics and Biopharmaceutics. 58(3),2004: 477-482.
57. Elsayed A, Remawi M A. Formulation and characterization of an oily-based system for oral delivery of insulin[J]. European Journal of Pharmaceutics and Biopharmaceutics. 73(2),2009:269-279.
58. Lior Z E, Kidron O M. Absorption of protein via the intestinal wall[J]. Biochemical Pharmacology, 36(7), 1987: 1035-1039.
59. Yamamoto A, Taniguchi T, Rikyuu K, et al.. Effects of various protease inhibitors on the intestinal absorption and degradation of insulin in rates[J]. Pharmaceutical Research. 11, 1994: 1496-1500.
60. Djordjevic L, Primorac M, Stupar M. In vitro release of diclofenac diethylamine from caprylocaproyl macrogolglycerides based microemulsions, International Journal of Pharmaceutics, 2005(296):73-79.
61. Sanggu Kim, Wai Kiong Nga, Shoucang Shen, et al.. Phase behavior, microstructure transition, and antiradical activity of sucrose laurate/propylene glycol/the essential oil of Melaleuca alternifolia/water microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 348(1), 2009: 289-297.
62. Moniruzzaman M, Kamiya N, Masahiro Goto. Ionic liquid based microemulsion with pharmaceutically accepted components: Formulation and potential applications[J]. Journal of Colloid and Interface Science, 2010, 352(1): 136-142.
63. Kogan A, Garti N. Microemulsions as transdermal drug delivery vehicles[J]. Advances in Colloid and Interface Science,2006,123-126:369–385.
64. Aramaki K, Ozawa K, Hironobu Kunieda, et al.. Effect of Temperature on the Phase Behavior of Ionic–Nonionic Microemulsions[J]. Journal of Colloid and Interface Science, 196(1), 1997: 74-78
65. Feng J L, Wang Z W, Juan Zhang. Study on food-grade vitamin E microemulsions basedon nonionic emulsifiers[J]. Colloids and Surfaces A: Physicochem. Eng, 339 (2009): 1-6.
66. Mehta S K, Bala K,, et al.. Tween-based microemulsions: a percolation view[J]. Fluid Phase Equilibria, 172(2), 2000: 197-209.
67. Warr G G, Shear and elongational rheology of ternary microemulsions[J], Colloids and Surfaces. 103, 1995: 273-279.
68. Palanuwech J, Potineni R, Robert F, et al.. A method to determine free fat in emulisons[J]. Food Hydrocolloids,2003,17(1):55-62.
69.江来,江荣高,薛艳.多肽和蛋白微乳制剂及其进展[J].天津药学.2007,19(3):66-69.
70. Charman S A, Charman W N, Rogg M C, et al.. Self-emulsifying systems formulation and biological evaluation of an investigative lipophilic compound [J]. Pharm Res, 1992, 9:87-94.
2. Hoar T P, Schulman J H. Transparent water-in-oil dispersions the oleopathic hydro-micelle [J]. Nature, 1943, 152: 102-03:21.
3. Jayne M L and Gareth D R. Microemulsion-based media as novel drug delivery systems[J]. Advanced Drug Delivery Reviews, 45(2000): 89-121.
4. Cilek, Celebi N, Tirnaksiz F, et al.. Tay A. A lecithin-based microemulsion of rh-insulin with aprotinin for oral administration: investigation of hypoglycemic effects in non-diabetic and STZ-induced diabetic rats[J]. International Journal of Pharmaceutics, 298(2005): 176-185.
5. Patel G, Misra A. Oral Delivery of Proteins and Peptides: Concepts and Applications[J]. Challenges in Delivery of Therapeutic Genomics and Proteomics, 2011: 481-529.
6. Tarver T. Food nanotechnology[J].Food Technology,2006,60(11):22-26
7. Chen H, Weiss J, Shahidi F, et al.. Nanotechnology in nutraceuticals and functional foods[J].Food Technology, 2006, 60(3):30-32.
8. Panayiotis P C, Scalart J P. Formulation and physical characterization of water in oil microemulsions containing long versus medium chain glycerides[J]. International Journal of Pharmaceutics, 158(1997):57-68
9. Guo-Rong Duan, Xu-Jie Yang, Guo-Hong Huang, Lu-De Lu. Water/span80/Triton X-100/n-hexyl alcohol/n-octane microemulsion system and the study of its application for preparing nanosized zirconia[J]. Materials Letters, 60(2006) 1582-1587.
10. Yiming Guo, Jingzhe Zhao, et al.. Preparation and characterization of monoclinic sulfur nanoparticles by water-in-oil microemulsions technique[J]. Powder Technology, 162 (2006):83-86.
11. Garti N, Aserin A, Fanun M. Non-ionic sucrose esters microemulsions for food applications. Part 1. Water solubilization[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 164(2000): 27-38.
12. Glatter O, Orthaber D, Stradner A, et al.. Sugar-Ester Nonionic Microemulsion: Structural Characterization[J]. Journal of Colloid and Interface Science, 241(2001):215-225.
13. Kreilgaard M, Pedersen E J, Jaroszewski J W. NMR characterisation and transdermal drug delivery potential of microemulsion systems[J]. Journal of Controlled Release,69(2000):421-433.
14. Sintova A C, Levy H V, Botner S, et al.. Systemic delivery of insulin via the nasal route using a new microemulsion system: In vitro and in vivo studies[J]. Journal of Controlled Release, 148(2010):168-176.
15. Graf A, Ablingerb E, Peters S, et al.. Microemulsions containing lecithin and sugar based surfactants Nanoparticle templates for delivery of proteins and peptides[J]. International Journal of Pharmaceutics, 350 (2008): 351-360.
16. Li-Ching Peng, Chi-Hsien Liub, Chang-Chin Kwana, Keh-Feng Huang. Optimization of water-in-oil nanoemulsions by mixed surfactants[J]. Colloids and Surfaces A: Physicochem. Eng. Aspects, 370 (2010):136-142.
17.路莹,吉小欣,高申,刘毅清.利多卡因-卵磷脂微乳处方的初步研究[J].药学实践杂志, 2004 22(3):141-143.
18. Alany R G, Rades T, Kustrin S A, et al.. Effects of alcohols and diols on the phase behaviour of quaternary systems[J]. International Journal of Pharmaceutics, 196(2000): 141-145.
19. Warisnoicharoen W, Lansley A B, Lawrence M J. Nonionic oil-in-water microemulsions: the effect of oil type on phase behaviour[J]. Int J Pharm, 2000(198):7-27.
20. Attwood D. Microemulsions [J]: Colloidal Drug Delivery Systems, Dekker, New 3171.
21. Constantinides P P, Seang H Y, et al.. Particle size determination of phase-inverted water-in-oil microemulsions under different dilution and storage conditions[J]. International Journal of Pharmaceutics, 115(1995):225-234.
22. Lutz Schlicht, Jan-Hendrik Spilgies, Georg Ilgenfritz. Dynamics of Structure Changes in Water-in-Oil Microemulsions: Electric Birefringence and Electric Light Scattering in Percolating Systems[J]. Journal of Molecular Liquids, 72(1997):295-314.
23. Fernandez J C, Bisceglia M, Acosta E, et al.. On the birefringence of water-alkane-AOT microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 157(1999):35-46.
24. Acosta E, Kurlat D H, Bisceglia M. Induced electric birefringence and viscosity studies in microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 106(1996):11-21.
25. Eicke H F, Hilfiker R. Connectivity-controlled charge transport in water-in-oil microemulsions[J]. Chemical Physics Letters, 1986,125(3):295-298.
26. Rozieres J, Marcia A. Middleton and Robert S, et al.. Schechter.Theoretical model for the complex permittivity of water-in-oil microemulsions with ionic surfactants[J]. Journal of Colloid and Interface Science, 1988 124(3):407-415.
27. Senatra D. Dielectric analysis and Differential Scanning Calorimetry of water-in-oil microemulsions[J]. Advances in Colloid and Interface Science, 123-126 (2006):415-424.
28. Senatra D, Ziparo C, Cecilia M. C. Gambi1, L. Lanzi, ENERGY BALANCE IN DENSE MICROEMULSIONS[J]. Journal of Thermal Analysis and Calorimetry, 2008 92 (2): 535-541.
29. Senatral D, Zhoul Z, Pieraccini L, et al.. A study of the properties of water-in-oil microemulsions in the subzero temperature range by differential scanning calorimetry[J]. Progr Colloid & Polymer Sci, 73(1987):66-75.
30. Garti N, Aserin A, Tiunova I, Fanun M. A DSC study of water behavior in water-in-oil microemulsions stabilized by sucrose esters and butanol[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 170(2000) 1-18.
31. Senatra D. Procedures for DSC analysis of percolative microemulsions[J]. Thermochimica Acta, 345 (2000) 39-46.
32. Campo L, Yaghmur A, et al.. Five-component food-grade microemulsions: structural characterization by SANS[J]. Journal of Colloid and Interface Science, 274 (2004):251–267.
33. Azouza B, Oberb R, Nakache E, Williams C E. A small angle X-ray scattering investigation of the structure of a ternary water-in-oil microemulsion[J]. Colloids and Surfaces, 1992,69(2-3): 87-97.
34. Behera G B, Mishra B K, Behera P K, Panda M. Fluorescent probes for structural and distance effect studies in micelles, reversed micelles and microemulsions[J]. Advances in Colloid and Interface Science, 82 (1999):1-42.
35. Nornoo A O, Zheng H, Lopes L B, et al.. Oral microemulsions of paclitaxel: In situ and pharmacokinetic studies[J]. European Journal of Pharmaceutics and Biopharmaceutics, 71(2009):310–317.
36. Walderhaug H. Microstructures in Aqueous Solutions of a Polyoxyethylene Trisiloxane Surfactant and a Cosurfactant Studied by SANS and NMR Self-Diffusion[J]. Langmuir, 2008(24):10637-10645.
37. Mehta S K, Kaur G, Bhasin K K, et al.. Tween-Embedded Microemulsions Physicochemical and Spectroscopic Analysis for Antitubercular Drugs[J]. AAPS PharmSciTech, 2010(11):143-153.
38. Fanun M. A study of the properties of mixed nonionic surfactants microemulsions by NMR, SAXS, viscosity and conductivity[J]. Journal of Molecular Liquids, 142(2008): 103-110.
39. Fanun M, Wail S A. Electrical conductivity and self diffusion NMR studies of the system:Water/sucrose laurate/ethoxylated monodiglyceride/isopropylmyristate[J]. Colloids and Surfaces A: Physicochem. Eng. Aspects, 277 (2006):83-89.
40. Zuliang Lai, Peiyi Wu. Investigation on the conformations of AOT in water-in-oil microemulsions using 2D-ATR-FTIR correlation spectroscopy[J]. Journal of Molecular Structure, 883-884 (2008):236-241.
41.梁文平,乳状液科学与技术基础[M],北京:科学出版社,2001,211-245.
42. Guptaa R R, Swantrant K. Jainb, Manoj Varshney, et al.. AOT water-in-oil microemulsions as a penetration enhancer in transdermal drug delivery of 5-?uorouracil[J]. Colloids and Surfaces B: Biointerfaces, 41 (2005):25-32.
43. Dracha M, Micha?eka J N, Sienkiewicz A. Antioxidative properties of vitamins C and E in micellar systems and in microemulsions[OL]. Colloids and Surfaces A: Physicochem. Eng, 2010.
44. Avramiotis S, Papadimitriou V, Cazianis C T, A. Xenakis. EPR studies of proteolytic enzymes in microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 144(1998):295-304.
45. Sharma G, Wilson K, Walle C F, et al.. Microemulsions for oral delivery of insulin: Design, development and evaluation in streptozotocin induced diabetic rats[J]. European Journal of Pharmaceutics and Biopharmaceutics, 76 (2010): 159-169.
46. Graf A, Rades T, Hook S M. Oral insulin delivery using nanoparticles based on microemulsions with different structure-types: Optimisation and in vivo evaluation[J]. european journal of pharmaceutical sciences, 37 (2009): 53-61.
47. Actis G C, Lagge M, Rizzetto M, et al.. Long-term efficacy of oral microemulsion cyclosporin for refractory ulcerative colitis[J].Minerva lVled,2004,95(1): 65-70.
48. Peira E, Scolari P, Gasco M R. Transdermal permeation of apomorphine through hairless mouse skin from microemulsions[J]. International Journal of Pharmaceutics, 226 (2001): 47-51.
49.王建磊,王正武,俞惠新.胰岛素W/O型微乳液的制备及体外释药性能[J].精细化学2006,23(9): 867-872.
50. Mao-Bo Cheng, Jian-Cheng Wang, Yu-Hua Li, Characterization of water-in-oil microemulsion for oral delivery of earthworm fibrinolytic enzyme[J]. Journal of Controlled Release, 129, 2008: 41-48.
51.韩旻,傅韶,方晓玲.三七总皂苷油包水微乳的处方筛选及体内外评价[J].药学学报2007,42(7): 780-786.
52.孟博宇,平其能,熊净,高琴.胰岛素微乳大鼠肠道给药降糖作用的研究[J].中国药科大学学报.2006,37(4): 304-307.
53. Lyons K C, Charman W N, Miller M, et al. Factors limiting the oral bioavailability of N-acetylglucosaminyl-N-acetylmuramyl dipeptide (GMDP) and enhancement of absorption in rats by delivery in a water-in-oil microemulsion[J]. International Journal of Pharmaceutics, 199 (2000): 17-28.
54. Damge C, Maincent P. Oral delivery of insulin associated to polymeric nanoparticles in diabetic mice[J]. Journal of Controlled Release.117. 2007:163–170.
55. Zelihagül Degim, Nilayünal. The effect of various liposome formulations on insulin[J]. Life Sciences. 75(23),2004: 2819-2827.
56. Cournarie F, Savelli M P. Insulin-loaded W/O/W multiple emulsions [J]. European Journal of Pharmaceutics and Biopharmaceutics. 58(3),2004: 477-482.
57. Elsayed A, Remawi M A. Formulation and characterization of an oily-based system for oral delivery of insulin[J]. European Journal of Pharmaceutics and Biopharmaceutics. 73(2),2009:269-279.
58. Lior Z E, Kidron O M. Absorption of protein via the intestinal wall[J]. Biochemical Pharmacology, 36(7), 1987: 1035-1039.
59. Yamamoto A, Taniguchi T, Rikyuu K, et al.. Effects of various protease inhibitors on the intestinal absorption and degradation of insulin in rates[J]. Pharmaceutical Research. 11, 1994: 1496-1500.
60. Djordjevic L, Primorac M, Stupar M. In vitro release of diclofenac diethylamine from caprylocaproyl macrogolglycerides based microemulsions, International Journal of Pharmaceutics, 2005(296):73-79.
61. Sanggu Kim, Wai Kiong Nga, Shoucang Shen, et al.. Phase behavior, microstructure transition, and antiradical activity of sucrose laurate/propylene glycol/the essential oil of Melaleuca alternifolia/water microemulsions[J]. Colloids and Surfaces A: Physicochemical and Engineering Aspects, 348(1), 2009: 289-297.
62. Moniruzzaman M, Kamiya N, Masahiro Goto. Ionic liquid based microemulsion with pharmaceutically accepted components: Formulation and potential applications[J]. Journal of Colloid and Interface Science, 2010, 352(1): 136-142.
63. Kogan A, Garti N. Microemulsions as transdermal drug delivery vehicles[J]. Advances in Colloid and Interface Science,2006,123-126:369–385.
64. Aramaki K, Ozawa K, Hironobu Kunieda, et al.. Effect of Temperature on the Phase Behavior of Ionic–Nonionic Microemulsions[J]. Journal of Colloid and Interface Science, 196(1), 1997: 74-78
65. Feng J L, Wang Z W, Juan Zhang. Study on food-grade vitamin E microemulsions basedon nonionic emulsifiers[J]. Colloids and Surfaces A: Physicochem. Eng, 339 (2009): 1-6.
66. Mehta S K, Bala K,, et al.. Tween-based microemulsions: a percolation view[J]. Fluid Phase Equilibria, 172(2), 2000: 197-209.
67. Warr G G, Shear and elongational rheology of ternary microemulsions[J], Colloids and Surfaces. 103, 1995: 273-279.
68. Palanuwech J, Potineni R, Robert F, et al.. A method to determine free fat in emulisons[J]. Food Hydrocolloids,2003,17(1):55-62.
69.江来,江荣高,薛艳.多肽和蛋白微乳制剂及其进展[J].天津药学.2007,19(3):66-69.
70. Charman S A, Charman W N, Rogg M C, et al.. Self-emulsifying systems formulation and biological evaluation of an investigative lipophilic compound [J]. Pharm Res, 1992, 9:87-94.