• Wantida Chaiyana Department of Pharmaceutical Science, Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand
  • Rungsinee Phongpradist Department of Pharmaceutical Science, Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand.
  • Pimporn Leelapornpisid Department of Pharmaceutical Science, Faculty of Pharmacy, Chiang Mai University, Chiang Mai 50200, Thailand.


Objective: The present study aims to investigate the compositions and biological activities of essential oil extracted from pomelo peel and develop into microemulsions.

Methods: Four subspecies of pomelo including Kao-Namphung (KN), Kao-Puang (KP), Kao-Tang-Gwa (KT), and Kao-Yai (KY) were subjected to the hydrodistillation to yield essential oils. The constituents of each oil was analyzed by GC-MS. Radical scavenging activities were determined by ABTS and DPPH assays, whereas, lipid antioxidant activity was determined by linoleic acid peroxidation assay. Antityrosinase activity and safety on human PBMCs were also investigated. Pseudoternary phase diagrams were constructed to reveal the effects of each compostions on the microemulsion regions. The microemulsion was formulated and chracterized for the particle size, rheological behavior and biological activities.

Results: Limonene was the major constituent in KN, KP, KT, and KY oil which was detected up to 86.19%, 85.76%, 79.36%, and 80.20%, respectively. Among four oils, KT oil exhibited the highest radical scavenging, antioxidant and antityrosinase activities. The MTT assay revealed that KT oil had no toxicity on human PBMCs. The microemulsion formulation (ME) containing 15% KT, 36% Tween 20, 9% PEG 400, and 40% water, were formulated and chracterized. ME was transparent liquid with the particle size of 90.28 ± 1.60 nm. ME exhibited the Newtonian flow behavior with low viscosity (16.78 ± 0.12 Pas). In a comparison with KT oil, ME show significant higher radical scavenging and antioxidant activities (p< 0.01).

Conclusion: Development of microemulsion increased radical scavenging and antioxidant activities of KT oil and would be an attractive system for further development to effective topical products.


Keywords: Pomelo, Essential oil, Antioxidant, Antityrosinase, Cytotoxicity, Microemulsion, Pseudoternary phase diagram.


Download data is not yet available.


1. Moulik SP, Paul BK. Structure, dynamics and transport properties of microemulsions. Adv Colloid Interface Sci 1998;78(2):99-195.
2. Lawrence MJ, Rees GD. Microemulsion-based media as novel drug delivery systems. Adv Drug Deliv Rev 2000;45(1):89-121.
3. Garcı́a-Sánchez F, Eliosa-Jiménez G, Salas-Padrón A, Hernández-Garduza O, Ápam-Martı́nez D. Modeling of microemulsion phase diagrams from excess Gibbs energy models. Chem Eng J 2001;84(3):257-74.
4. Dixit SG, Mahadeshwar AR, Haram SK. Some aspects of the role of surfactants in the formation of nanoparticles. Colloids Surf A Physicochem Eng Asp 1998;133(1):69-75.
5. Prince LM. A theory of aqueous emulsions I. Negative interfacial tension at the oil/water interface. J Colloid Interf Sci 1967;23(2):165-73.
6. Gasco MR. Microemulsions in the pharmaceutical field: perspectives and applications. Surfactant Sciser1997;66:97-122.
7. Špiclin P, Homar M, Zupančič-Valant A, Gašperlin M. Sodium ascorbyl phosphate in topical microemulsions. Int J Pharm 2003;256(1):65-73.
8. Chen H, Chang X, Du D, Li J, Xu H, Yang X. Microemulsion-based hydrogel formulation of ibuprofen for topical delivery. Int J Pharm 2006;315(1):52-8.
9. Akanabe Y, Kusunoki A, Ikeda Y, Tanaka M. A comparison of volatile components from the peel of ohshima no. 1 with its parent cultivars. Biosci Biotechnol Biochem 2008;72(7):1969-72.
10. Jena SN, Kumar S, Nair NK. Molecular phylogeny in Indian Citrus L. (Rutaceae) inferred through PCR-RFLP and trnL-trnF sequence data of chloroplast DNA. Sci Hort 2009;119(4):403-16.
11. Moore GA. Oranges and lemons: clues to the taxonomy of Citrus from molecular markers. Trends Genet 2001;17(9):536-40.
12. Al-Bahrany AM. Effect of phytohormones on in vitro shoot multiplication and rooting of lime Citrus aurantifolia (Christm.) Swing Sci Hort 2002;95(4):285-95.
13. Arias BÁ, Ramón-Laca L. Pharmacological properties of citrus and their ancient and medieval uses in the Mediterranean region. J Ethnopharmacol 2005:97(1):89-95.
14. Lim HK, Moon JY, Kim H, Cho M, Cho SK. Induction of apoptosis in U937 human leukaemia cells by the hexane fraction of an extract of immature Citrus grandis Osbeck fruits. Food Chem 2009;114(4):1245-50.
15. Mokbel MS, Hashinaga F. Evaluation of the antioxidant activity of extracts from buntan (Citrus grandis Osbeck) fruit tissues. Food Chem 2006;94(4):529-34.
16. Darah I, Taufiq MMJ, Lim SH. Pomelo Citrus grandis (L.) osbeck peel as an economical alternative substrate for fungal pectinase production. Food Sci Biotechnol 2013;22(6):1683-90.
17. Knight TG, Klieber A, Sedgley M. The relationship between oil gland and fruit development in Washington navel orange (Citrus sinensis L. Osbeck). Ann Botany 2001:88(6):1039-47.
18. Siripongvutikorn S, Thummaratwasik P, Huang YW. Antimicrobial and antioxidation effects of Thai seasoning, Tom-Yum. LWT-Food Sci Technol 2005;38(4):347-52.
19. Saeio K, Chaiyana W, Okonogi S. Antityrosinase and antioxidant activities of essential oils of edible Thai plants. Drug Discov Ther 2011;5(3):144-9.
20. Adams RP. Quadrupole mass spectra of compounds listed in order of their retention time on DB-5. Identification of essential oils components by gas chromatography/quadrupole mass spectroscopy. Allured Publishing Co, Carol Stream, IL, USA; 2001.
21. Fellegrini N, Ke R, Yang M, Rice-Evans C. Screening of dietary carotenoids and carotenoid-rich fruit extracts for antioxidant activities applying 2, 2′-azinobis (3-ethylenebenzothiazoline-6-sulfonic acid radical cationdecolorization assay. Meth enzymol 1999;299:379-89.
22. Blois MS. Antioxidant determination by the use of a stable free radical. Nature 1958;181:1199-200.
23. Niehius WG, Samuelson B. Formation of malondialdehyde from phospholipid arachido-nate during microsomal lipid peroxidation. Eur J Biochem 1968;6(1):126-30.
24. Pomerantz SH. Separation, Purification, and properties of two tyrosinases from hamster melanoma. J Biol Chem 1963;238(7):2351-7.
25. Phongpradist R, Chittasupho C, Okonogi S, Siahaan T, Anuchapreeda S, Ampasavate C, et al. LFA-1 on leukemic cells as a target for therapy or drug delivery. Curr Pharm Des 2010;16(21):2321-30.
26. Chaiyana W, Saeio K, Hennink WE, Okonogi S. Characterization of potent anticholinesterase plant oil based microemulsion. Int J Pharm 2010;401(1):32-40.
27. Njoroge SM, Koaze H, Karanja PN, Sawamura M. Volatile constituents of redblush grapefruit (Citrus paradisi) and pummelo (Citrus grandis) peel essential oils from Kenya. J Agric Food Chem 2005;53(25):9790-4.
28. Hosni K, Zahed N, Chrif R, Abid I, Medfei W, Kallel M, et al. Composition of peel essential oils from four selected tunisian citrus species: evidence for the genotypic influence. Food Chem 2010;123(4):1098–104.
29. Janaszewska A, Bartosz G. Assay of total antioxidant capacity: comparison of four methods as applied to human blood plasma. Scand J Clin Lab Inv 2002;62(3):231-6.
30. Bauzaite R, Venscutonis PR, Gruzdiene D, Tirzite D, Tirzitis G. Radical scavenging and antioxidant activity of various plants grown in Lithuania. Food Technology and Quality Evaluation: Science Publishers, In3, USA; 2003.
31. Moon JK, Shibamoto T. Antioxidant assays for plant and food components. J Agric Food Chem 2009;57(3):1655-66.
32. Tirzitis G, Bartosz G. Determination of antiradical and antioxidant activity: basic principles and new insights. Acta Biochim Pol 2010;57:139-42.
33. Halliwell B, Chirico S. Lipid peroxidation: its mechanism, measurement, and significance. Amer J Clin Nutr 1993:57(5):715S-24S.
34. Niki E, Yoshida Y, Saito Y, Noguchi N. Lipid peroxidation: mechanisms, inhibition, and biological effects. Biochem Biophys Res Commun 2005;338(1):668-76.
35. Candan F, Unlu M, Tepe B, Daferera D, Polissiou M, Sökmen A, et al. Antioxidant and antimicrobial activity of the essential oil and methanol extracts of Achilleamillefolium subsp. Millefolium Afan. (Asteraceae). J Ethnopharmacol 2003;87(2):215-20.
36. Abirami A, Nagarani G, Siddhuraju P. In vitro antioxidant, anti-diabetic, cholinesterase and tyrosinase inhibitory potential of fresh juice from Citrus hystrix and C. maxima fruits. Food Sci Human Wellness 2014;3:16-25.
37. Wu SJ, Ng CC, Tzeng WS, Ho KC, Shyu YT. Functional antioxidant and tyrosinase inhibitory properties of extracts of Taiwanese pummelo (Citrus grandis Osbeck). Afr J Biotechnol 2013;10(1):7668-74.
38. Kreilgaard M. Influence of microemulsions on cutaneous drug delivery. Adv Drug Deliv Rev 2002;54:S77-S98.
39. Ali AA, Mulley BA. Formation of liquid crystal and other non‐fluid phases in emulsions containing non‐ionic surfactants. J Pharm Pharmacol 1978;30(1):205-13.
40. Groves MJ, Ahmad AB. Some rheological properties of lyotropic liquid crystals formed by phosphate polyoxyethylene surfactants, n-hexane and water. Rheol Acta 1976;15(9):501-7.
41. Alany RG, Rades T, Agatonovic-Kustrin S, Davies NM, Tucker IG. Effects of alcohols and diols on the phase behaviour of quaternary systems. Int J Pharm 2000;196(2):141-5.
42. Garti N, Aserin A, Ezrahi S, Wachtel E. Water solubilization and chain length compatibility in nonionic microemulsions. J Colloid Interf Sci 1995;169(2):428-36.
43. Gao ZG, Choi HG, Shin HJ, Park KM, Lim SJ, Hwang KJ, et al. Physicochemical characterization and evaluation of a microemulsion system for oral delivery of cyclosporin A. Int J Pharm 1998;161(2):75-86.
44. Kale NJ, Allen Jr LV. Studies on microemulsions using Brij 96 as surfactant and glycerin, ethylene glycol and propylene glycol as cosurfactants. Int J Pharm 1989;57(2):87-93.
45. Krauel K, Girvan L, Hook S, Rades T. Characterisation of colloidal drug delivery systems from the naked eye to Cryo-FESEM. Micron 2007;38(8):796-803.
46. Boonme P, Krauel K, Graf A, Rades T, Junyaprasert VB. Characterization of microemulsion structures in the pseudoternary phase diagram of isopropyl Palmitate/Water/Brij 97:1-Butanol. AAPS Pharm Sci Tech 2006;7(2):99-104.
47. Chaiyana W, Rades T, Okonogi S. Characterization and in vitro permeation study of microemulsions and liquid crystalline systems containing the anticholinesterase alkaloidal extract from Tabernaemontanadivaricata. Int J Pharm 2013;452(1):201-10.
48. Okonogi S, Chaiyana W. Enhancement of anti-cholinesterase activity of Zingibercassumunar essential oil using a microemulsion technique. Drug Discov Ther 2012;6(5):249-55.
745 Views | 2743 Downloads
How to Cite
Chaiyana, W., R. Phongpradist, and P. Leelapornpisid. “CHARACTERIZATION OF HYDRODISTILLATED POMELO PEEL OIL AND THE ENHANCEMENT OF BIOLOGICAL ACTIVITIES USING MICROEMULSION FORMULATIONS”. International Journal of Pharmacy and Pharmaceutical Sciences, Vol. 6, no. 9, 1, pp. 596-02,
Original Article(s)