CHEMICAL COMPOSITION, ANTIOXIDANT, ANTIBACTERIAL AND CYTOTOXICITY ANALYSIS OF BLUMEA LACERA (BURM. F.) DC Original Article SAVITA KHATRI, NEETU PHOUGAT, RENU CHAUDHARY, BHARAT SINGH, ANIL KUMAR CHHILLAR* Centre for Biotechnology, Maharshi Dayanand University, Rohtak, Haryana, India Email: anil.chhillar@gmail.com Received: 06 May 2016 Revised and Accepted: 20 Jun 2016 ABSTRACT Objective: To investigate the in vitro antibacterial and antioxidant potential of B. lacera (leaves). The most active extracts were examined for their chemical composition and cytotoxicity. Methods: The antibacterial activity of five different extracts were examined against 8 bacterial strains (5 Gram-positive and 3 Gram-negative) using resazurin based microtitre dilution assay (RMDA) and disk diffusion assay (DDA). The antioxidant activity of extracts was demonstrated by using DPPH (1, 1-diphenyl-2-picrylhydrazyl) assay and superoxide radical scavenging assay. Chemical composition and cytotoxicity were assessed by using gas chromatography-mass spectrometry (GC-MS) and haemolytic assay, respectively. Results: According to RMDA, the petroleum ether extract (PEE) and chloroform extract (CE) exhibited highest antibacterial activity. The PEE showed highest activity against Salmonella enterica ser. typhi and Serratia marcescens with MIC i.e. 390.62 μg/ml. Similarly, the CE showed highest antibacterial activity against Bacillus cereus and Micrococcus luteus with MIC i.e. 390.62 μg/ml. In DPPH assay, CE showed the highest radical scavenging activity with IC50 Conclusion: The present investigation represents B. lacera as an incredible herb. The PEE and CE were found to possess promising antibacterial and antioxidant properties. The CE exhibited lesser toxicity as compared with PEE. 57.46 µg/ml. In GCMS analysis, the principal compounds in PEE and CE were stigmasterol (12.86 %) and L-(+)-ascorbic acid 2, 6-dihexadecanoate (11.73 %), respectively. In haemolytic assay, the PEE and CE showed non-toxic behaviour up to 125 µg/ml and 500 µg/ml, respectively. Keywords: Antibacterial, Antioxidant, B. lacera, Chemical composition, Cytotoxicity © 2016 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/) INTRODUCTION Multidrug resistance to conventional antibiotics is responsible for the alarming rates of pathogenic microbes. Bacterial infections are still amongst the leading causes of mortality worldwide [1]. Imprudent use of chemotherapeutic agents and improper diagnosis of microbial infections leads to the emergence of drug resistant pathogens [2]. This imperative need for novel therapeutic agents leads to re-emergence of natural products for drug discovery. According to Food and Drug Administration (FDA), 34% of new approved medicines between 1981 and 2010 including anticancer drugs and immunosuppressants were based on natural products or their derivatives [3]. Phytoconstituents are also a safer alternative to prevent cancers [4]. Oxidative stress and cell death are widely related to the generation of reactive oxygen species (ROS) which attack on macromolecules. Various degenerative human diseases including Alzheimer’s disease, diabetes mellitus, neurodegenerative disorders, Parkinson’s disease, inflammation, atherosclerosis, cancer and ageing are associated with uncontrolled generation of free radicals [5]. Several medicinal plants are known for their antioxidant capacity since antiquity [6]. The genus Blumea consists of about 80 species. B. lacera (Burm. f.) DC (Compositae) is generally known as Janglimulli, Siyalmutra, Kakaronda and Susksampatra [7]. It is a perennial, corymbosely branched herb with well known odour of camphor. Its stem is erect with ash colour. The whole plant is covered with long soft hairs. Leaves are obovate and deeply serrated. It has yellow groundsel-like flowers, arranged in axillary cymes or terminal panicle. Fruits are oblong and without ribs. Its blooming period is during January to April. It is generally found in roadside areas, river margins and wastelands. B. lacera occurs all over the plains of India from the north-west ascending to 2,000 ft in the Himalayas. It is also dispersed to Australia, China and Tropical Africa. Ethnomedicinally, the plant is used as an astringent, stimulant, thermogenic, styptic, opthalmic, digestive, liver tonic, anthelmintic and expectorant [8]. B. lacera also possesses anticancer, anti-inflammatory, antispasmodic, antipyretic, tranquilizing and diuretic activities. Various phytoconstituents including flavones, triterpenes, β-sitosterol, cineol, campesterol, lupeol, hentriacontane and α-amyrin are obtained from essential oils, leaves, root and bark of B. lacera [9]. It is well established that essential oils of B. lacera possessed a broad range of activity spectrum however true potential of its leaves formulations has to be explored. So, we have carried out this study with a broad range of representative bacterial strains and investigate the in vitro antibacterial potential of B. lacera. The in vitro antioxidant potential of B. lacera was also evaluated. Chemical composition and cytotoxicity of extracts having antibacterial and antioxidant properties were assessed. MATERIALS AND METHODS Plant material Fresh leaves of B. lacera were collected from District Jhajjar, Haryana, India, in November 2012. The plant was identified from Department of Botany, Maharshi Dayanand University, Rohtak, Haryana (India) with voucher no. CBT-05 and further authenticated with the help of flora of Haryana [10]. Chemicals and reagents Petroleum ether, chloroform, acetone, methanol, dimethylsulfoxide (DMSO), hydrochloric acid, Dragendorff’s reagent, ferric chloride, conc. sulphuric acid, Luria broth (LB), Luria agar (LA), resazurin dye, gentamicin, 1, 1-diphenyl-2-picryl hydrazyl radical (DPPH), ascorbic acid, nitro blue tetrazolium (NBT), sodium dihydrogen orthophosphate, di-sodium hydrogen orthophosphate, sodium chloride, NADH, phenazine metho-sulphate (PMS), gallic acid, triton X-100 were purchased from Himedia Chemicals; India. Extraction procedure Fresh leaves were air-dried under shade for four successive weeks at room temperature. The dried plant material was chopped and International Journal of Pharmacy and Pharmaceutical Sciences ISSN- 0975-1491 Vol 8, Issue 8, 2016