Establishment of a method to detect sulfonamide residues in chicken meat and eggs by high-performance liquid chromatography J.M.K.J.K. Premarathne a, b, * , D.A. Satharasinghe a , A.R.C. Gunasena a , D.M.S. Munasinghe a , P. Abeynayake a a Faculty of Veterinary Medicine and Animal Science, University of Peradeniya, Peradeniya, 20400, Sri Lanka b Faculty of Livestock, Fisheries and Nutrition, Wayamba University of Sri Lanka, Makandura, 60170, Gonawila, Sri Lanka article info Article history: Received 15 May 2015 Received in revised form 21 November 2015 Accepted 11 December 2015 Available online xxx Keywords: Food safety Antimicrobial residues Sulfonamides abstract Sulfonamides are a group of antimicrobials used for treatment and prevention of infectious diseases in humans and animals. In veterinary practice, sulfonamides are extensively used due to its broad spectrum of activity and low cost. A multi-residue analysis of seven sulfonamides (sulfadiazine, sulfathiazole, sulfapyridine, sulfamerazine, sulfamethazine, sulfamethizole, and sulfamethoxypyridazine) in chicken and eggs using HPLC- DAD detection method has been proposed. Chicken and egg samples were ho- mogenized and extracted with distilled water:ethyl acetate (1:3, v/v) liquid-liquid extraction. The ex- tracts were defatted with n-heptane and dried under nitrogen flow at 55  C. The dry residue was dissolved in 500 mL methanol:acetic acid:water mixture with a ratio of 10:4:36 (v/v/v) and 10 mL of the sample was subjected to HPLC determination under the following conditions: column, Luna 5 m C18; particle size, 5 mm; mobile phase, 17 mM acetic acid: methanol: acetonitrile (83:10:7, v/v/v); flow rate, 1.0 mL/min; and detection, 270 nm. The specificity was evaluated by analyzing 30 different blank samples of chicken and eggs in order to verify the absence of potential interfering compounds. No interfering peaks were found around the retention time of analytes in the matrices under investigation. The linear correlation coefficients (r 2 ) for 50e250 ppb range were above 0.99 for all sulfonamides tested. The mean recoveries for chicken and eggs spiked at 50, 100, and 150 ppb were in the range of 86e108% for all analytes. Repeatability and within laboratory reproducibility of the developed method was determined at 100 ppb and quantified as the relative standard deviation was lower than 15%. The de- cision limits were between 108 and 116 ppb for all analytes whilst the detection capability of all analytes ranged from 129 to 140 ppb. An inexpensive and simple liquid-liquid extraction with isocratic elution mode for rapid analysis of residues of seven sulfonamides in chicken and egg samples using HPLC-DAD detection was established in this study. © 2015 Elsevier Ltd. All rights reserved. 1. Introduction Sulfonamides are a group of antimicrobials used for treatment and prevention of infections in humans and in animals (Bishop, 2005). In veterinary practice, sulfonamides (SAs) are extensively used due to their broad spectrum of activity and low cost (Saif et al., 2003). In addition, SAs are used as feed additives to promote growth in animals (Bishop, 2005). Improper use of sulfonamides can lead to the development of resistant microorganisms and the occurrence of residues in animal origin food products (Giguere et al., 2006). Sulfadiazine (SDZ) has been shown to have toxic effects especially on the thyroid (Tsai, Kondo, Ueyama, & Azama, 1995). Therefore to ensure consumer safety, the European Union (EU) has set the maximum residue limits (MRLs) of 100 ppb for sulfonamides in food of animal origin. Various physiochemical and immunological methods have been developed to detect and quantify sulfonamide residues in animal origin food products. Among them are single or simultaneous analysis of sulfonamides by immunoassay methods (De Keizera, Bienenmann-Plouma, Bergwerff, & Haasnoota, 2008; Zhang et al., 2007) or biosensor immunoassay (Haasnoot, Bienenmann-Ploum, & Kohen, 2003; Situ, Crooks, Baxter, Ferguson, & Elliottet, 2002), multi-residue analysis of sulfonamides using liquid * Corresponding author. Faculty of Livestock, Fisheries and Nutrition, Wayamba University of Sri Lanka, Makandura, 60170, Gonawila, Sri Lanka. E-mail address: krissjayaruk@yahoo.com (J.M.K.J.K. Premarathne). Contents lists available at ScienceDirect Food Control journal homepage: www.elsevier.com/locate/foodcont http://dx.doi.org/10.1016/j.foodcont.2015.12.012 0956-7135/© 2015 Elsevier Ltd. All rights reserved. Food Control xxx (2015) 1e7 Please cite this article in press as: Premarathne, J. M. K. J. K., et al., Establishment of a method to detect sulfonamide residues in chicken meat and eggs by high-performance liquid chromatography, Food Control (2015), http://dx.doi.org/10.1016/j.foodcont.2015.12.012