e n v i r o n m e n t a l t o x i c o l o g y a n d p h a r m a c o l o g y 3 6 ( 2 0 1 3 ) 135–141 Available online at www.sciencedirect.com jo ur nal homepage: www.elsevier.com/locate/etap Genoprotective effects of lignin isolated from oil palm black liquor waste Prashantha Naik a , Hj Din Rozman b , Rajeev Bhat c,* a Department of Biosciences, Mangalore University, Mangalagangothri 574 199, India b Bioresources, Paper and Coatings Technology, School of Industrial Technology, Universit Sains Malaysia, Penang 11800, Malaysia c Food Technology Division, School of Industrial Technology, Universit Sains Malaysia, Penang 11800, Malaysia a r t i c l e i n f o Article history: Received 25 October 2012 Received in revised form 9 February 2013 Accepted 11 March 2013 Available online 18 March 2013 Keywords: Antioxidants Lignin Genotoxicity Cytotoxicity Cyclophosphamide Micronucleus test a b s t r a c t Black liquor waste (BLW), a major by-product of palm oil extraction process contains lignin as one of the constituents. Lignin isolated from BLW was evaluated for antioxidant and genoprotective properties and was compared with the commercial lignin for overall efficacy. Antioxidant compounds (phenolics and tannins) and antioxidant activities (phosphomolyb- denum assay, ABTS + and FRAP assays) of lignin isolated from BLW were compared with commercial lignin. Bone marrow micronucleus (MN) test was employed for evaluating the dose-yield protective effect against cyclophosphamide (CP, 50 mg/kg b.w.) induced geno- toxicity in mouse. Results revealed isolated lignin to exhibit rich antioxidant activities. A decrease in MN frequency and recovery of P/N ratio (P: polychromatic erythrocytes, N: nor- mochromatic erhythocytes) indicated protective effects of lignin against cyclophosphamide induced genotoxicity and cytotoxicity. The efficacy of BLW-derived lignin as an antioxidant and genoprotective agent was comparable to commercial lignin. Results on lignin isolated from BLW are envisaged to find potential applications in food and/or pharmaceutical indus- tries. © 2013 Elsevier B.V. All rights reserved. 1. Introduction Palm oil extraction process results in the generation of large quantity of wastes, which is of a major apprehension to surmount some of the major environmental issues. Today, researchers are constantly looking for the possible alternatives of utilizing the palm-based wastes (lingo-cellulose) for produc- tion of bio-fuels or using them as reinforcement or composite materials for developing non-toxic and easily biodegradable biopolymer packaging films. The major by-product of oil extraction process comprises of black liquor waste (BLW). This BLW is an aqueous solution consisting of lignin and hemi- celluloses along with the residues of inorganic chemicals ∗ Corresponding author. Tel.: +60 4653 5212; fax: +60 4657 3678. E-mail addresses: rajeevbhat1304@gmail.com, rajeevbhat@usm.my (R. Bhat). used during the extraction process. BLW, if left untreated, can results in generation of stinking smell (due to microbial inter- actions) thus polluting the environment (Bhat et al., 2009). Lignin, being a natural polyphenolic compound is non- toxic, and is reported to exhibit antimicrobial, anti-parasitic, antioxidant, anticancer, genoprotective and immuno- protective activities (Mikulásová and Kosíková, 2003; Mitjans and Vinardell, 2005; Bhat et al., 2009). Previously, we have reported similarity in the functional groups between commer- cial lignin and lignin isolated from BLW, and how this can be an effective low cost replacement for commercial lignin (Bhat et al., 2009). In view of various therapeutic values of lignin, the present investigation was undertaken to evaluate antioxidant 1382-6689/$ – see front matter © 2013 Elsevier B.V. All rights reserved. http://dx.doi.org/10.1016/j.etap.2013.03.004