Development and stability evaluation of Olive oil nanoemulsion using Sucrose ester Ahmad M M Eid, Rosnani Hasham, Mariani AbdulHamid, Farahdiana bt Ariffin, Nagib A. Elmarzugi Institute of Bioproduct Development, Universiti Teknologi Malaysia, 81310 UTM Johor Bahru, Malaysia, email: dr_ahmad98@hotmail.com Introduction Nanoemulsions have uniform and extremely small droplet sizes, typically in the range of 20–200 nm (Maestro et al, 2008). Its high kinetic stability, low viscosity and optical transparency make them very attractive systems for many industrial applications (Taha et al., 2004). Nanoemulsions have also been reported as one of the most promising techniques for enhancement of transdermal permeation and bioavailability of poorly soluble drugs (Baboota, S., et al., 2007). Preparation of nanoemulsion can be achieved by using two methods either high or low energy emulsification method. High energy emulsification method can be obtained by using high pressure homogenizers aided by appropriate choice of surfactants and co-surfactants. However, low energy emulsification method can be achieved by application of phase inversion temperature (Tharwat et al., 2004). Olive oil is considerably rich in monounsaturated fats, mostly oleic acid and high content of antioxidative substances, which linked with a reduction in the risk of coronary heart disease (Keys et al., 1986). Also Anti-aging properties, moisturizing the skin and anti-inflammatory properties. That make it a good candidate for nanoemulsion. The main of this study is to prepare olive oil nanoemulsion using low energy emulsification method. Formulation of nanoemulsion by adding water to Pre-nanoemulsion. Pre-nanoemulsion was prepared by mixing of sucrose laurate and glycerin then oily part added until complete dispersion occur under heat (80 ⁰C). Droplet Size Figure 1. represent ternary phase diagrams for three different combinations of oil, surfactant and glycerol. Ternary phase diagram was constructed to find the optimum composition of oil, surfactant and glycerol that could produce nanoemulsion. It can be seen in the ternary phase diagrams, different areas that correspond to different emulsions, such as transparent nanoemulsion (NE), macroemulsion (ME), coarse emulsion (CE) and very fast separated. Conclusion: Pre-emulsion was prepared using sucrose laurate, olive oil and glycerol by mixing the ingredients under heat. The best concentrations of oil can be used were 50% and 60% with 25% sucrose laurate. After achieving pre-emulsion it was diluted with water under gentile agitation to produce olive oil nanoemulsion. References: - Baboota S., Shakeel F., Ahuja A., Ali J. & Shafiq S. Design development and evaluation of novel nanoemulsions formulations for transdermal potential of celecoxib. Acta Pharm 8, 316-332 (2007). - Keys A., Menotti A., Karvonen M.J., et al. The diet and 15-year death rate in the Seven Countries Study. Am J Epidemiol 124, 903-915 (1986). - Taha E I., Al-saidan S., Samy A M. & Khan M A. Preparation and in-vitro characterization of self-nanoemulsified drug d elivery system (SNEDDS) of all trans-retinol acetate, Int. J. Pharm 285(1-2): 109-19 (2004). - Tharwat T P., Izquierdo J. & Esquena C S. Formation and stability of nano- emulsions. Journal of Colloid and Interface Science 108-109, 303-318 (2004). - Maestro A., Solé I., González C., Solans C. & Gutiérrez J.M. Influence of the phase behavior on the properties of ionic nanoemulsions prepared by the phase inversion composition method. Journal of Colloid and Interface Science 327, 433-439 (2008). METHOD RESULT AND DISCUSSION Figure 1. Ternary phase diagram of olive oil nanoemulsion. CONCLUSION Droplet Size Stability Olive oil Sucrose Laurate Glycerol Formulation Olive oil Laurate 1695 Particle size (nm) PDI F1 50% 10% 272 ± 2.38 0.321 F2 50% 15% 251 ± 2.08 0.409 F3 50% 20% 190 ± 2.52 0.339 F4 50% 25% 156 ± 3.21 0.251 F5 60% 10% 186 ± 2.67 0.499 F6 60% 15% 179 ± 2.34 0.419 F7 60% 20% 169 ± 1.73 0.336 F8 60% 25% 143 ± 1.54 0.285 Table 1. The results of droplet size and size distribution of 50% and 60% olive oil nanoemulsion. Table 1. Shows that the mean droplet size was decreased with an increased in surfactant concentration from 10% to 25% for both 50% and 60% olive oil nanoemulsion formulations. F4 and F8 show the smallest drolplets and the lowest PDI. Formulation Observation Initial After 1 month 4 0 C 25 0 C 40 0 C F5 Droplet size 186 ± 2.67 194 ± 4.63 203 ± 4.53 472 ± 5.12 PDI 0.499 0.480 0.662 0.739 F6 Droplet size 179 ± 2.34 198 ± 3.89 216 ± 4.69 425 ± 4.14 PDI 0.419 0.449 0.510 0.670 F7 Droplet size 169 ± 1.73 192 ± 2.32 214 ± 3.19 336 ± 3.19 PDI 0.336 0.394 0.482 0.661 F8 Droplet size 143 ±1.54 157 ± 3.69 169 ± 3.98 356 ± 4.12 PDI 0.285 0.290 0.308 0.332 After 3 month 4 0 C 25 0 C 40 0 C 220 ± 4.54 275 ± 4.21 585 ± 3.56 0.401 0.562 0.598 215 ± 4.45 278 ± 5.25 509 ± 3.4 0.569 0.698 0.645 217 ± 2.04 235 ± 4.7 465 ± 3.7 0.43 0.432 0.79 197 ± 3.45 233 ± 4.73 434 ± 2.72 0.3 0.371 0.475 After 3 month 4 0 C 25 0 C 40 0 C 280 ± 2.8 387 ± 6.65 575 ± 3.42 0.395 0.42 0.598 285 ± 5.51 328 ± 5.25 559 ± 4.14 0.569 0.517 0.645 207 ± 4.04 255 ± 3.87 565 ± 2.3 0.43 0.432 0.69 180 ± 2.65 233 ± 4.73 443 ± 3.2 0.35 0.371 0.575 Formulation Observation Initial After 1 month 4 0 C 25 0 C 40 0 C F1 Droplet size 272 ± 2.38 278 ± 2.62 358 ± 3.65 475 ± 2.42 PDI 0.321 0.375 0.386 0.493 F2 Droplet size 251 ± 2.08 265 ± 5.51 290 ± 4.51 459 ± 6.14 PDI 0.409 0.569 0.387 0.431 F3 Droplet size 190 ± 2.52 207 ± 4.04 215 ± 3.72 365 ± 2.31 PDI 0.339 0.43 0.374 0.429 F4 Droplet size 156 ± 3.21 167 ± 2.65 173 ± 2.43 345 ± 2.12 PDI 0.251 0.28 0.271 0.411 Table 3. The stability results of 60% droplet size and uniformity at 4 0 C, 25 0 C and 40 0 C. Table 2. The stability results of 50% droplet size and uniformity at 4 0 C, 25 0 C and 40 0 C. Based on the result obtained in table 1, statistical result showed that there were no significant difference (p > 0.05) on droplet size for 50% olive oil pre-nanoemulsion containing 25% sucrose laurate after 1 month storage at 4 0 C and 25 0 C, while after 3 months stored at 25 0 C shows significant difference but there is no difference at 4 0 C. However, there was significant difference on droplet size for formulation stored at 40 0 C after 1 and 3 months. Based on the result obtained in table 2, statistical result showed that there were no significant difference (p > 0.05) on droplet size for 50% olive oil nanoemulsion containing 25% sucrose laurate after 1 month storage at 4 0 C and 25 0 C, while after 3 months nanoemulsion stored at 25 0 C shows significant difference but there is no difference at 4 0 C. However, there was significant difference on droplet size for formulation stored at 40 0 C after 1 and 3 months. Formulation Droplet size (nm) Zeta potential F4 173.7 ± 1.43 -64.4 ± 1.40 F8 164.6 ± 1.49 -67.2 ± 1.52 Table 4. Shows results of droplet size and zeta potential of optimum nanoemulsion. F4 is a mixture of pre-nanoemulsion formulation 50% oil and 25% Laureate. While F8 is a mixture of 60% oil and 25% Laureate to produce nanoemulsion. Results show the F4 and F8 were in nano-size with high negative zeta potential which indicate that the nanoemulsions were very stable. Table 4. Results of droplet size and zeta potential of nanoemulsion. Material and Method Olive oil, Sucrose Monoester Laurate and glycerol were used to prepare the nanoemulsion. First Pr-emulsion was prepared by mixing oil, Laureate and glycerol under heat (80 ⁰C). Then nanoemulsion was prepared by self-emulsifying the pre-emulsion with water under gentle agitation.