• Rachmawati Ramadhana Mustofa Laboratory of Physical Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.
  • Iskandarsyah . Laboratory of Physical Pharmacy, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia.


Objective: This study aimed to prepare and characterize anti-acne ethosomes using the cold- and thin-layer hydration methods.
Methods: A sonication step was included during ethosome preparation to improve the quality of the cold method. Azelaic acid, Phospholipon 90G,
ethanol, propylene glycol, and phosphate buffer (pH 7.4) were used in the procedures. Prepared ethosomal suspensions were characterized using
transmission electron microscopy, particle-size analysis, and spectrophotometry.
Results: Ethosomes prepared using the thin-layer hydration method (F1) had small unilamellar vesicles, while those prepared using the cold method
with 15-min sonication (F4) showed spherical, elliptical, unilamellar, and multilamellar vesicles. F1 ethosomes had a Dmean volume of 648.57±231.26,
whereas those prepared using the cold method with 5- (F2), 10- (F3), and 15-min (F4) sonication had Dmean volumes of 2734.04±231.49 nm,
948.90±394.52 nm, and 931.69±471.84 nm, respectively. Polydispersity indices of F2, F3, and F4 ethosomes were 0.74±0.21, 0.86±0.05, and 0.91±0.03,
respectively, with a poor particle-size distribution, compared to that of F1 (0.39±0.01). Zeta potentials of F1–F4 ethosomes were −38.27±1.72 mV,
−23.53±1.04 mV, −31.4±1.04 mV, and −34.3±1.61 mV, respectively. Entrapment efficiencies of F1–F4 ethosomes were 90.71±0.11%, 53.84±3.16%,
72.56±0.28%, and 75.11±1.42%, respectively.
Conclusion: Anti-acne ethosomes produced using the thin-layer hydration method had superior properties than those produced using the cold
method with 15-min sonication.

Keywords: Azelaic acid, Characterization, Cold method, Ethosome, Preparation, Sonication, Thin-layer hydration method.


1. Manosroi J, Apriyani MG, Foe K, Manosroi A. Enhancement of the
release of azelaic acid through the synthetic membranes by inclusion
complex formation with hydroxypropyl-beta-cyclodextrin. Int J Pharm
2. O’Neil M. The Merck Index. 14th ed. Whitehouse Station (NJ): Merck;
2006. p. 153.
3. Ascenso A, Raposo S, Batista C, Cardoso P, Mendes T, Praça FG,
et al. Development, characterization, and skin delivery studies of
related ultradeformable vesicles: Transfersomes, ethosomes, and
transethosomes. Int J Nanomedicine 2015;10:5837-51.
4. Shen S, Liu SZ, Zhang YS, Du MB, Liang AH, Song LH, et al.
Compound antimalarial ethosomal cataplasm: Preparation, evaluation,
and mechanism of penetration enhancement. Int J Nanomedicine
5. Mistry A, Ravikumar P. Development and evaluation of azelaic acid
based ethosomes for topical delivery for the treatment of acne. Indian J
Pharm Educ Res 2016;50:232-43.
6. Mbah C, Builders P, Nzekwe I, Kunle O, Adikwu M, Attama A.
Formulation and in vitro evaluation of pH-responsive ethosomes
for vaginal delivery of metronidazole. J Drug Deliv Sci Technol
7. Celia C, Cilurzo F, Trapasso E, Cosco D, Fresta M, Paolino D,
et al. Ethosomes® and transfersomes® containing linoleic acid:
Physicochemical and technological features of topical drug delivery
carriers for the potential treatment of melasma disorders. Biomed
Microdevices 2012;14:119-30.
8. Maestrelli F, Capasso G, González-Rodríguez ML, Rabasco AM,
Ghelardini C, Mura P, et al. Effect of preparation technique on the
properties and in vivo efficacy of benzocaine-loaded ethosomes.
J Liposome Res 2009;19:253-60.
9. Chen JG, Liu YF, Gao TW. Preparation and anti-inflammatory activity
of triptolide ethosomes in an erythema model. J Liposome Res
10. Wirarti GA. Formulations of Green Tea Leaves (Camellia sinensis L.
Kuntze) Etosomic Gel Preparations and in vitro Penetration Test
Using Franz Diffusion Cells. Depok: Universitas Indonesia; 2016.
p. 26-7.
11. Abdulbaqi IM, Darwis Y, Khan NA, Assi RA, Khan AA. Ethosomal
nanocarriers: The impact of constituents and formulation techniques
on ethosomal properties, in vivo studies, and clinical trials. Int J
Nanomedicine 2016;11:2279-304.
12. Kadam T, Darekar A, Gondkar S, Saudagar R. Development and
validation of spectrophotometric method for determination of azelaic
acid. Asian J Res Pharm Sci 2015;5:83-5.
13. Li G, Fan Y, Fan C, Li X, Wang X, Li M, et al. Tacrolimus-loaded
ethosomes: Physicochemical characterization and in vivo evaluation.
Eur J Pharm Biopharm 2012;82:49-57.
14. Smith E, Dea PK. Applications of calorimetry in a wide contextdifferential
scanning calorimetry, isothermal titration calorimetry
and microcalorimetry. Differential Scanning Calorimetry Studies
of Phospholipid Membranes: The Interdigitated Gel Phase. Ch. 18.
London: InTech; 2013.
15. Alberts B, Johnson A, Lewis J, Raff M, Roberts K, Walker P. Molecular
Biology of the Cell. 4th ed. New York: Garland Science; 2002.
16. Prasanthi D, Lakshmi PK. Vesicles-mechanism of transdermal
permeation: A review. Asian J Pharm Clin Res 2012;5:18-25.
17. Nurleni N, Iskandarsyah I, Aulia A. Formulation and penetration
testing of ethosome azelaic acid on abdominal skin white male rats
(Rattus Norvegicus) with franz diffusion cell. Asian J Pharm Clin Res
18. Nimisha S, Srivastava K, Singh AK. Formulation and evaluation of sea
buckthron leaf extract loaded ethosomal gel. Asian J Pharm Clin Res
19. Suslick KS. Kirk-Othmer Encyclopedia of Chemical Technology.
4th ed. New York: J. Wiley and Sons; 1998. p. 517-41.
20. Malvern Instruments. Dynamic Light Scattering Common Terms
Defined. Malvern Instruments; 2011. p. 1-6.
21. Marto J, Vitor C, Guerreiro A, Severino C, Eleutério C, Ascenso A,
et al. Ethosomes for enhanced skin delivery of griseofulvin. Colloids
Surf B Biointerfaces 2016;146:616-23.
35 Views | 68 Downloads
How to Cite
Mustofa, R. R., & ., I. (2018). PREPARATION AND CHARACTERIZATION OF ANTI-ACNE ETHOSOMES USING COLD AND THIN-LAYER HYDRATION METHODS. International Journal of Applied Pharmaceutics, 10(1), 338-342.
Original Article(s)