Research Article ANTIOXIDANT AND ANTIMICROBIAL ACTIVITY OF PHYLLANTHUS EMBLICA FOR ITS APPLICATION IN TREATMENT OF OPHTHALMIC DISORDERS S.S.SAWHNEY*, R.M.PAINULI**, DOLLY SINGH** * Uttaranchal College of Science and Technology, Dehradun, India, ** H.N.B. Garhwal Central University, Srinagar, Garhwal, India. Email: dolly.ms@gmail.com Received: 13 June 2011, Revised and Accepted: 3 July 2011 ABSTRACT The oxidation stress is known to be a reason for various tissue degeneration and plays a role in premature aging. In wake of this, there is need to explore a natural antioxidant which can help body maintain a balance between inbuilt oxidation stress and detoxifying reactions. We present the antioxidant power of various extracts of Phyllanthus emblica which was tested by FRAP method. The phytochemical analysis of the extracts was checked by HPLC. Along with the standard compound, gallic acid, the peaks were analyzed. The peaks showed the compound present was mainly, Gallic acid. The antioxidant activity was found to be maximum in the order of Ethyl Acetate extract followed by Acetone, Methanol and Aqueous extract. Along with antioxidant activity the extracts were tested as an anti‐microbial agent for the pathogens known to cause various eye infections. The minimal inhibitory concentration was calculated and was found to be 1mg/ml for S.mutans, S.aureus and S.pyogenes and 0.6mg/ml for S.pneumoniae. Phyllanthus emblica is a powerful antioxidant along with its activity to inhibit the growth of the pathogens makes us believe that it can be used as new drug compound for preparation of eye drops and also in various therapeutic. Keywords: Phyllanthus emblica, HPLC, Antioxidant, Eye infection, Broad‐spectrum antibiotic, MIC INTRODUCTION Oxidative Stress (OS) is a generally used to describe the sturdy state of oxidative damage in a cell or tissue and can also effect the normal function of organs which is caused by the reactive oxygen species (ROS) 1 . This oxidative stress affects either a specific molecule or the organism as a whole. Reactive oxygen species represent a class of molecules that are derived from the metabolism of oxygen and are inherently found in all aerobic organisms in form of free radicals and peroxides 2 . Energy‐derived metabolic activity or endogenous source are the primary source of production of such free radicals such as those involving detoxification reactions like liver cytochrome P‐450 enzyme system or the energy generation from mitochondria. Enzyme system or the energy generation from mitochondria 3 . Beside these endogenous source the environmental factors that play role in ROS generation includes exposure to cigarette smoke, emission from automobiles and industries, consumption of alcohol in excess, and bacterial, viral or fungal infection 4 . The exogenous factors beside the environmental pollutants are vitamins A, C, E, carotenoids etc 5 . Oxidative stress is resultant of an imbalance between production of the reactive oxygen and a body's ability to naturally detoxify the reactive intermediates. Resultant of this imbalance oxidation stress is involved in most of the major disorders like cardiovascular diseases, arthrosclerosis and Parkinson’s and in the minor imbalance results in premature ageing 6 . In serious diseases like, Duchenne Muscular Dystrophy (DMD), progressive muscle cell necrosis occurs due to loss of dystrophin and its demonstrated in mice that muscle cells, under oxidative stress are severely damaged which is generally protected by Stra13 and in absences of Stra13 muscle necrosis progress leading to symptoms of DMD in mice 7 . The level of damage due to oxidative stress is determined by the balance between induced oxidative damage versus the rate at which it is capably repaired and removed by the natural system 8 . Damage can also be determined by the rate of reactive oxygen generated and how rapidly the endogenous agents inactivate ROS like antioxidants and the level of repair enzymes present in the human body in turn decides this. There is involvement of the individual hereditary factors that are unique for each human and these can act as one of determinant factors in fighting the oxidation stress built up in the body. Besides these factors the lifestyle and environment of the each individual can also play a key role in the fighting ability of the body against the oxidation stress at any given point 9 . Present day clinical treatments are having a setback because of the ever‐evolving nature of microorganisms, which have developed resistance to them, and hence treatment of infectious diseases is problematic scenario 10 . The infections caused by bacteria, viruses and fungus can also lead to the built up of the reactive oxygen species in the body 11 . Oxidative damage is majorly observed in the eyes particularly affecting the retina and the lens, is an also known contributing factor to age‐related ocular degeneration and cataract 12 . The present study in detail describes the extraction procedure, phytochemical analysis, antioxidant power and screening of the antimicrobial activity of Phyllanthus emblica against pathogens especially known to cause various eye infections. MATERIAL AND METHODS Materials The plants material (seeds) was collected from the local market in Mumbai. Mueller‐Hinton agar was purchased from Hi‐Media, Mumbai, Bacterial Inoculum was procured from IMTECH chandigarh, India. Chemicals used for HPLC analysis were of HPLC grade. The microorganisms used in this study are Streptococcus mutans (S.mutans), Streptococcus pyogenes (S.pyogenes), Streptococcus pneumoniae (S.pneumoniae), Staphylococcus aureus (S.aureus) and Pseudomonas aeruginosa (P.aeruginosa). These microorganisms were procured from the IMTECH, Chandigarh, INDIA. Preparation of plant extracts Dried fruits were powdered in the grinder and stored in the airtight container in the dark until further use. 25g of the powder of seeds of P.emblica were taken in a thimble and the extracts were prepared in the series of 500ml of different solvents based on increasing polarity (Ethyl acetate, Acetone, Methanol, and Water) using Soxhlet extraction method. The solvents were evaporated to dryness using rotary evaporator and the extracts were lyophilized at –50°C. The lyophilized extracts powder was stored in airtight bottles at 4°C till further experimentation 13 . High Performance Liquid Chromotagraphy (HPLC) HPLC analysis was carried out using C18 PCX 500 Dionex analytical column, (WATER 2414 refractive index detector and 515 HPLC pump) with 0.1 M KCl, 0.05 M HCl and 10% acetonitrile as the mobile phase. The detection was carried out at 260nm using UV detector. Peak areas were identified, and gallic acid was used as an external standard 14 . International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 3, Issue 4, 2011 A Ac c a a d d e e m mi i c c S Sc c i i e e n n c c e e s s