Vol 13, Issue 8, 2020 Online - 2455-3891 Print - 0974-2441 EVALUATION OF IN VITRO ANTI-INFLAMMATORY ACTIVITY OF CITRUS MACROPTERA MONTR SONAM BHUTIA* Department of Pharmacognosy, Government Pharmacy College, Government of Sikkim, Sikkim University, Rumtek, Sajong, East Sikkim. Email: sonamkzbhutia@gmail.com Received: 28 April 2020, Revised and Accepted: 09 June 2020 ABSTRACT Objective: The use of naturally occurring medicines dependent on essential oils (EOs) is nowadays of great interest. In addition, within the human body, EO shows high efficacy as antioxidants and anti-inflammatory drugs. The present experiment was conducted to access the anti-inflammatory activity of EO obtained from the fruit peels of Citrus macroptera Montr. (Rutaceae) against the denaturation of protein in vitro model. Methods: The test sample (EO) was incubated under controlled laboratory conditions at varying concentrations with egg albumin and was subjected to absorbance determination for the anti-inflammatory property analysis. Diclofenac sodium was used as the standard reference drug for the experiment. Results: The results show a concentration-dependent inhibition of protein (albumin) denaturation by the test oil. This was concluded by comparing their IC 50 average values. Citrus macroptera Montr. EO possessed IC 50 average value 54.6+0.07 μg/mL, whereas that of diclofenac sodium was found to be 52.89+0.06 μg/ml. The result shows that the test oil is more effective than the standard drug. Conclusion: From the above experimental finding, it can be concluded that Citrus macroptera Montr. EO has significance anti-inflammatory effect against the denaturation of the protein in vitro model. The activity may be due to the presence of terpene polyphenolic component or some other active compound present in the oil. The provided information was first of its kind of knowledge to keep the scientific data for future reference. Key words: Anti-inflammatory, essential oil, Citrus macroptera, Protein denaturation, Polyphenolic compounds. INTRODUCTION Inflammation is a complex process often associated with pain and involves occurrences such as increased vascular permeability, enhanced protein denaturation, and rearrangement of the membrane. When cells in the body are damaged by microbes, physical agents, or chemical agents, the injury is in the form of stress. Inflammation of tissue is due to response to stress. It is a defensive response that is characterized by redness, pain, heat, and swelling and loss of function in the injured area. Loss of function depends on the area and severity of the condition occurring. Since inflammation is one of the non-specific intelligences which is based on mechanisms of the body, a tissue response to an unintentional cut is similar to that of other forms of tissue damage caused by heat, radiation, bacterial, or viral [1]. An organism or tissue elicits the inflammatory responses as a defensive mechanism; however, prolonged inflammation can result in undesired health effects as a result of interplaying various biomolecules that are secreted during the inflammation phase. Inflammation in many diseases has been documented including cancer [2]. Chronic pain induced by inflammatory processes is a major clinical problem worldwide, steroidal and nonsteroidal anti- inflammatory drugs (NSAIDs) are the most widely used treatments in these chronic pain states. NSAIDs, such as diclofenac, aspirin, and indometacin, block the biosynthesis pathway of prostaglandins by inhibiting the cyclooxygenase (COX) enzymes, producing anti-inflammatory, analgesic, and antipyretic effects [3]. At present, used medications for pain control and inflammatory disorders pose dangerous side effects on chronic administration. Attempts are, therefore, being made to research promising plants which can lead to the production of newer or safer drugs [4]. MATERIALS AND METHODS Collection, identification, and authentication of plant The fruits of the plant Citrus macroptera were collected locally from East Khasi Hills, Meghalaya. The plant was identified, confirmed, and authenticated by Botanical Survey of India, Shillong, Meghalaya. Extraction of EO The fruit peels of Citrus macroptera were thoroughly washed with distilled water, cut into small pieces and about 200 g in four fractions were subjected to hydrodistillation using a Clevenger apparatus for about 4 h. The steamed and vaporized oil were condensed into liquid by a vertical condenser and collected in a measuring cylinder. Being immiscible and lighter than water, the volatile oil is separated out as an upper layer. The oil from the fruit peels were collected by following the standard procedure (Chutia et al., 2009). Finally, it was dried over anhydrous sodium sulfate and kept in an airtight container at 4–8°C until further analysis (Isman, 2000). Standard drugs Diclofenac sodium was purchased from Sigma-Aldrich Co., and purity was labeled to be 98%, respectively. Preparation of crystallized egg albumin The crystallized albumin obtained from 24 eggs was dissolved in about 500 ml of distilled water and poured into 5 L of boiling distilled water with rapid stirring so that the coagulated albumin was finely divided. It was then poured on several large filters and washed until the washings were entirely free of sulfates. The substance was then allowed to drain thoroughly, transferred to a flask containing about 10 volumes of 95% boiling alcohol for an hour, and filtered. This was repeated once with five volumes of 95% alcohol and once with five volumes of absolute alcohol. The material was filtered and then stirred with three volumes of a good grade of ether and again filtered. This process was repeated twice and the protein was finally dried in a vacuum desiccator over sulfuric acid. This gave a dry white product which could easily be reduced to a powder. The yield was about 1 g/egg [5-7]. © 2020 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/ajpcr.2020.v13i8.38063 Research Article