ISSN - 0975-7058 Vol 10, Special Issue 1, 2018 ISSN - 0975-7058 NOVEL TRANSETHOSOME CONTAINING GREEN TEA (CAMELLIA SINENSIS L. KUNTZE) LEAF EXTRACT FOR ENHANCED SKIN DELIVERY OF EPIGALLOCATECHIN GALLATE: FORMULATION AND IN VITRO PENETRATION TEST EFFIONORA ANWAR*, DELLY RAMADON, GHINA DESVIYANTI ARDI Department of Pharmaceutical Technology, Faculty of Pharmacy, Universitas Indonesia, Depok, 16424, Indonesia. Email: effionora.anwar@farmasi.ui.ac.id Received: 20 June 2018, Revised and Accepted: 19 November 2018 ABSTRACT Objective: This study aimed to formulate a transethosome cream (TEC) to increase skin penetration of epigallocatechin gallate (EGCG) in green tea leaf extract and evaluate their physicochemical characteristics and skin penetration capacity. Methods:Transethosomeswerepreparedthroughthin-layerhydrationmethodinthreeformulations(F1−F3)withdifferentSpan80andethanolconcentrations. Transethosome morphology was characterized using transmission electron microscopy, particle size, polydispersity (PDI), and zeta potential using a particle size analyzer and entrapment efficiency (EE). Penetration was tested using an in vitro Franz diffusion cell with female Sprague-Dawley rat skin as the membrane. Results: Transethosome F2 containing green tea extract equivalent to 3% EGCG, 4% Lipoid P30, 0.75% Span 80, and 30% ethanol had the best characteristic including spherical shape, smallest particle size (35.35 nm), 0.319 PDI, zeta potential of −29.97±3.05 mV, and highest EE of 45.26%±8.15%. TEC permitted greater flux than non-TEC (first phase: 60.56±4.52 vs. 25.69±0.83 µg•cm −2 •h −1 and second phase: 23.13±1.38 vs. 7.36±1.59 µg•cm −2 •h −1 ). Conclusion: Transethosome can increase the skin penetration of green tea leaf extract. Keywords: Cream, Epigallocatechin gallate, Green tea leaf extract, Penetration, Transethosome. INTRODUCTION Green tea from the tea plant Camellia sinensis has several health benefits [1]. Bioactive compounds present in green tea leaves include alkaloids, flavonoids, and phenols [2]. The flavonoids (particularly the catechin fraction) are basic phenolic compounds responsible for antioxidant activity [3]. Epigallocatechin gallate (EGCG) is the major catechin in green tea and the primary source of its antioxidant activity [4]. EGCG is hydrophilic (log p=0.48) [5] and thus has poor skin permeability. However, skin delivery has several advantages such as the mitigation of gastrointestinal disturbances and bypass of hepatic first- pass metabolism [6]. Therefore, a delivery system that can facilitate the skin absorption of EGCG is desirable, and a formulation using transethosomes could be among the best such vehicles for EGCG-based treatment. For the skin delivery of such compounds, several strategies have been used to bypass the stratum corneum barrier. Liposomes have been used for topical drug delivery; however, conventional liposomes do not deeply penetrate the skin [7]. To overcome this limitation, novel lipid vesicles - transethosome and transfer some - were developed. Transethosome is a modified ethosome formed into lipid vesicles that can improve penetration through the skin [8]. Transethosome contains high ethanol content together with an edge activator or permeation enhancer [7]. Transethosome can be formulated into a semi-solid form such as a cream. Creams generally have greater esthetic appeal because they are non-greasy and can be easily applied, spread, and removed [9]. In this study, transethosomes containing green tea extract were formulated into a cream (transethosome cream, TEC) and evaluated for their physicochemical characteristics and skin penetration capacity. This formulation exhibited highly advantageous physicochemical characteristics, high entrapment efficiency (EE), and markedly enhanced skin flux than a non-TEC (NTEC). METHODS Materials Green tea leaf extract (C. sinensis L. Kuntze) (Andy Biotech [Xi’an] Co. Ltd., China), EGCG standard (Sigma-Aldrich, Singapore), Lipoid P30 (Lipoid GmbH, Germany), solvents, and other chemical reagents were of analytical grade. EGCG assay of green tea leaf extract Green tea left extract was prepared according to the methods of Fangueiro et al. with some modifications [10]. Standard solutions of EGCG were prepared at 25, 30, 35, 40, 45, and 50 µg/mL for constructing a calibration curve. The solution was analyzed using reversed-phase- high-performance liquid chromatography with a UV-visible detector set at 280 nm and a flow rate of 1.0 mL/min. The mobile phase comprised 0.05% (v/v) acetic acid: acetonitrile (87:13 v/v, pH approximately 4.0). The injection volume was 20 µg/mL, and retention time was approximately 16 min. Antioxidant activity of green tea leaf extract The antioxidant activity was measured according to the method of Mishra et al [11]. Ascorbic acid standard and green tea leaf extract solutions were prepared at different concentrations. DPPH 100 µg/mL (1.0 mL) and methanol (2.0 mL) were added to 1.0 mL of each sample solution. The mixture was shaken and incubated at room temperature for 30 min. The antioxidant activity was determined using a UV-visible spectrophotometer at 514 nm. Research Article © 2018 The Authors. Published by Innovare Academic Sciences Pvt Ltd. This is an open access article under the CC BY license (http://creativecommons. org/licenses/by/4. 0/) DOI: http://dx.doi.org/10.22159/ijap.2018.v10s1.66 The 2 nd Physics and Technologies in Medicine and Dentistry Symposium (PTMDS), Universitas Indonesia. Depok, Indonesia