Abstract—Anthocyanins are natural pigments with effective UV protection but their topical use could be limited due to their physicochemical characteristics. An attempt to overcome such limitations by complexation of 2 major anthocyanin-rich sources, C. ternatea and Z. mays, has potentiated its use as topical anti- inflammatory. Cell studies indicate no cytotoxicity of the anthocyanin complex (AC) up to 1 mg/ml tested in HaCaT and human fore head fibroblasts by MTT. Croton oil-induced ear edema in Wistar rats suggests an effective dose of 5 mg/cm 2 of AC as a topical anti-inflammatory in comparison to 0.5 mg/cm 2 of fluocinolone acetonide. Niosomal encapsulation of the AC significantly prolonged the anti-inflammatory activity particularly at 8 h after topical application (p = 0.0001). The AC was not cytotoxic and its anti-inflammatory and activity was dose-dependent and prolonged by niosomal encapsulation. It has also shown to promote collagen type 1 production in cell culture. Thus, AC could be a potential candidate for topical anti-inflammatory agent from natural resources. Keywords—Anthocyanin complex, ear edema, inflammation, niosomes, skin. I. INTRODUCTION NTHOCYANINS are natural pigments daily intake from coloured fruits, flowers, and vegetables [1]. Anthocyanins possess anti-inflammatory activity [2], [3], anti- oxidant [4], [5], anti-carcinogenic [6] and anti-microbial including anti-candida activities [7], [8]. Topical application of anthocyanins has been reported for UV protection on the skin [9], however, the use of anthocyanins can be restricted due to physicochemical charateristics including pH, colour staining and poor skin permeation. Complex formation of natural anthocyanins is common in plants so as to physicochemically stabilize the molecular conformation of anthocyanins [10], [11]. Complex formation of natural anthocyanins is common in plants so as to physicochemically stabilize the molecular conformation of anthocyanins [10], [11]. Niosomes are self-assembly vesicles which mimic bilayer structure of cell membrane, and could be used to enhance skin permeation of the encapsulated compounds [12]. An anthocyanin complex (AC) formed with 2 major A. Priprem is with the Division of Pharmaceutical Technology, Faculty of Pharmaceutical Science, Khon Kaen University, 123 Mittraphap Rd, KhonKaen 40002, Thailand (Corresponding author: Tel.: +66-43-362092; fax: +66-43-202379. E-mail apriprem@gmail.com). S. Limsitthichaikoon is with the Division of Pharmaceutical Technology, Faculty of Pharmaceutical Science, Khon Kaen University, 123 Mitraparb Rd, Muang, KhonKaen, 40002, Thailand (e-mail: lsucharat@gmail.com). S. Thappasarapong is with the Division of Pharmaceutical Chemistry, Faculty of Pharmaceutical Science, Khon Kaen University, Khon Kaen 40002, Thailand (e-mail:sutpit1@kku.ac.th). anthocyanins, i.e. delphinidin- and cyanidin-rich sources, was able to be taken up by one of the human fibroblast cells [13]. Since fibroblasts play a vital role in the viable skin, including responses to inflammatory stimuli [14]. Forming a complex might retard skin permeation, thus an in vivo test on the effectiveness of topical applications of the anthocyanin complex is required. In this light, niosomal encapsulation was introduced for comparison on the permeation of anthocyanin complex. Topical steroids are commonly prescribed as effective anti- inflammatory property, however, there are many common side effects and complications occurred such as steroid allergy, steroid-induced skin atrophy, including secondary infection from bacteria, virus and fungus, if long-term or high dose uses [15]. If the anthocyanin complex has been shown to be effective as a topical anti-inflammatory, and also the niosomes could encapsulate the anthocyanin complex, a potential use of the anthocyanins from abundant natural resources is made possible. II. MATERIALS AND METHODS A. Anthocyanin Complex (AC) Preparation Dried blue flowers of Clitoria ternatea L. (Sisaket, Thailand), identified [16] and herbarium collected no.KKUAP_03 at the institute plant collection, were extracted by water and freeze-dried to obtain its crude extract (CT). Dried cobs of purple waxy corn (Zea mays L. ceritina Kulesh.) from an open-pollinated variety (Kao Kum) harvested in the university campus were grinded and extracted by water followed by freeze-dried to obtain its crude extract (CC). An anthocyanin complex (AC) was formed using the previously described protocol [13]. In brief, an aqueous mix of CC, CT and turmeric rhizome (ratio = 5:5:1) was heated to 80ºC and trace amount of Zn 2+ (Ajax Finechem, Australia) and caffeic acid (Sigma-Aldrich, China) were mixed and then cooled to -20°C to precipitate. AC powder was obtained from the oven-dried precipitate. B. Preparations of AC AC solution was prepared by dissolving AC powder with 0.1% sodium polyacrylate (GMP, Bangkok, Thailand) in isotonic phosphate buffer saline at pH 7.4. Span 60 (Sigma, St. Louis, MO, U.S.A.) and cholesterol (Sigma, St. Louis, MO, U.S.A.) were dissolved in 1:1 chloroform and methanol and rotary evaporated under vacuum to form a thin film. The thin film was rehydrated with the AC solution to form AC niosomes. Blank niosomes were prepared Anti-Inflammatory Activity of Topical Anthocyanins by Complexation and Niosomal Encapsulation Aroonsri Priprem, Sucharat Limsitthichaikoon, Suttasinee Thappasarapong A World Academy of Science, Engineering and Technology International Journal of Chemical and Molecular Engineering Vol:9, No:2, 2015 142 International Scholarly and Scientific Research & Innovation 9(2) 2015 scholar.waset.org/1307-6892/10000421 International Science Index, Chemical and Molecular Engineering Vol:9, No:2, 2015 waset.org/Publication/10000421