~ 95 ~ International Journal of Herbal Medicine 2014; 2 (2): 95-99 ISSN 2321-2187 IJHM 2014; 2 (2): 95-99 Received: 22-07-2014 Accepted: 30-07-2014 Caroline Mathen OCT Therapies and Research Pvt. Ltd., 1 st F loor, Devendra M achinery & Fabricators Compound, Tank Road, Bhandup W est, Mumbai 400 078, India Renuka Thergaonkar K ETs V .G. V aze College, Department of P ost Graduate D iploma in Perfumery and Cosmetics M anagement, M ulund East, M umbai 400 082 Moushami Teredesai K ETs V .G. V aze College, Department of P ost Graduate D iploma in Perfumery and Cosmetics M anagement, M ulund East, M umbai 400 082 Gaurav Soman National Centre for Cell Science, NCCS Complex, University of Pune Campus, Ganeshkhind, Pune 411007, M aharashtra, India Shiney Peter St. X avier’s College, Department of Biotechnology, 5, M ahapalika M arg, M umbai, M aharashtra 400 001 Correspondence: Caroline Mathen OCT Therapies and R esearch Pvt. Ltd., 1 st Floor, Devendra Machinery & Fabricators Compound, Tank Road, Bhandup West, Mumbai 400 078, India Evaluation of anti-elastase and antioxidant activity in antiaging formulations containing terminalia extracts Caroline Mathen, Renuka Thergaonkar, Moushami Teredesai, Gaurav Soman and Shiney Peter ABSTRACT Most in vitro efficacy studies of creams are based on estimation of active constituents as solubility is the limiting factor. Our aim was to establish a method for efficacy testing of both anti-elastase and antioxidant activity in final formulations and to compare the same with the activity of the active ingredients to ascertain whether the native activity remains unaffected, enhanced or is compromised in any way. The actives evaluated in the antiaging formulations included the crude extracts of Terminalia arjuna and Terminalia chebula, either alone or in combination on the activity of porcine pancreatic elastase and to reduce 2, 2-Diphenyl-1-picrylhydrazyl (DPPH). The results showed that the T. arjuna formulation had higher inhibitory activity of elastase (IC50 6.6 mg/ml) while T. chebula formulation depicted better antioxidant activity (IC50 0.032 mg/ml). The combination cream had excellent anti- elastase (IC50 2.8 mg/ml) and antioxidant properties (IC50 0.14 mg/ml). Further the cytotoxicity of the two extracts was tested on 3T3 cell line and both extracts were found to have negligible inhibition up to 80 μg/ml. Keywords: anti-elastase, antioxidant, DPPH, antiaging, Terminalia arjuna, Terminalia chebula, cytotoxicity 1. Introduction The two distinct types of aging that exist may be due to genetic inheritance implicated in intrinsic (internal) involving cellular senescence and extrinsic (external) aging, such as photoaging [1, 2] . A consequence of both however is the production of free radicals at whose door the blame is laid for the loss of skin elasticity; one of the classical aging characteristics. Skin firmness and elasticity is mainly contributed to by elastin [3] , a dermal protein which is a constituent of the connective tissue (CT). Over time, the metabolism of the CT proteins slows down accompanied by an increase in enzymatic activity, particularly elastase, which breaks down elastin [4] . One way to prevent such a loss of elasticity is to use active ingredients that are able to inhibit these enzymes [5] and we concentrated on evaluation of the anti-elastase activity. Another molecule of interest is the antioxidant which functions by contributing an additional oxygen molecule to free radicals. In the absence of antioxidants, the free radicals will acquire an oxygen molecule from another biological molecule and this leads to tissue damage and eventually to skin aging. Kim and colleagues [6] , have described the various reactive oxygen species (ROS) which cause skin damage. For the most part, free radicals are chemically unstable and are a normal by-product of metabolic processes, but have a negative effect on the body as they are detrimental to cells. It is thus logical to incorporate compounds with free radical scavenging activity in an antiaging formulation [7] . We estimated antioxidant activity by the DPPH assay. DPPH is a stable free radical in methanol and is purple-coloured. The odd electron in DPPH becomes paired with hydrogen from a free radical scavenging antioxidant to form reduced DPPH-H. The resultant decolorization from purple to yellow can be detected spectro-photometrically. Various studies have already reported multi faceted biological activities of the Terminalia species, the anti-oxidant and anti-diabetic potential among the most commonly reported [8, 9, 10] . Others have rationalized that a formulation containing Terminalia extracts would be highly beneficial in an anti-aging and skin protective context [11, 12] . The majority of in vitro studies on anti-elastase and antioxidant activities are based on the evaluation of the actives. We incorporated the ethanolic extracts of T. arjuna and T. chebula in anti-aging formulations and