Sensors and Actuators B 199 (2014) 330–338 Contents lists available at ScienceDirect Sensors and Actuators B: Chemical jo u r nal homep age: www.elsevier.com/locate/snb Development of electrochemical biosensor with ceria–PANI core–shell nano-interface for the detection of histamine Manju Bhargavi Gumpu a,b , Noel Nesakumar a,c , Swaminathan Sethuraman a,b , Uma Maheswari Krishnan a,b , John Bosco Balaguru Rayappan a,c,∗ a Centre for Nanotechnology & Advanced Biomaterials (CeNTAB), SASTRA University, Thanjavur 613 401, Tamil Nadu, India b School of Chemical and Biotechnology, SASTRA University, Thanjavur 613 401, Tamil Nadu, India c School of Electrical & Electronics Engineering, SASTRA University, Thanjavur 613 401, Tamil Nadu, India a r t i c l e i n f o Article history: Received 27 January 2014 Received in revised form 19 March 2014 Accepted 3 April 2014 Available online 15 April 2014 Keywords: Histamine CeO2–PANI Diamine oxidase Cyclic volatammetry Amperometry Tiger prawn a b s t r a c t A mediator-free electrochemical biosensor with CeO 2 –PANI nano-interface for sensing histamine using diamine oxidase (DAO) has been developed. CeO 2 –PANI core–shell nanoparticles were prepared by hydrothermal method. The field emission scanning electron microscopy (FE-SEM) revealed the aggre- gated spherical morphology of CeO 2 . The core–shell formation of CeO 2 –PANI was confirmed with field emission transmission microscopy (FE-TEM). The polycrystallinity of CeO 2 and CeO 2 –PANI was confirmed using X-ray diffraction (XRD). Immobilization of DAO with CeO 2 –PANI was confirmed with Fourier trans- form infrared spectroscopy (FT-IR). Electrochemical studies were carried out through cyclic voltammetry and amperometry using modified GCE/CeO 2 –PANI/DAO as a working electrode, Ag/AgCl saturated with 0.1 M KCl as a reference electrode and platinum (Pt) wire as a counter electrode. The linear range was observed from 0.45 to 1.05 mM with a sensitivity of 724.94 A cm -2 mM -1 . Michaelis–Menten constant was calculated as 0.798 mM. It exhibited limit of detection of 48.7 M, limit of quantification of 132.4 M with a response time of <1 s and good shelf life of 86% till 18 days. Also the developed biosensor was applied on tiger prawn to estimate the histamine content. © 2014 Published by Elsevier B.V. 1. Introduction Histamine is a heterocyclic low molecular weight organic com- pound formed by enzymatic removal of carboxyl group from histidine, found in various foods such as meat, fish, alcoholic bever- ages, vinegar, fermented foods etc [1,2]. Improper handling, storage conditions, availability of free amino acids and optimum tempera- tures are the main reasons for increase in histamine concentration thereby leading to the toxification of fish related food items [3,4]. Consumption of such fish leads to “Histaminosis” or “Scombroid fish poisoning” causing both physiological and toxicological effects based on dosage [5]. According to World review of fisheries and aquaculture in 2011, production of fish is estimated to be 154 million tonnes all over the world which signifies that the rate of consumption is being increased. As a result, safety evaluation of fish and its products ∗ Corresponding author at: Centre for Nanotechnology & Advanced Biomaterials (CeNTAB) & School of Electrical & Electronics Engineering (SEEE) SASTRA University, Thanjavur 613 401, India. Tel.: +91 4362 264 101x255; fax: +91 4362 264120. E-mail address: rjbosco@ece.sastra.edu (J.B.B. Rayappan). became a key determinant to avoid adverse effects. As a part of food safety evaluation, levels of histamine in fish are limited to 0.45 mM by Food and Drug Administration (FDA) [3,5]. To detect histamine levels in fish and its related products, many enzymes such as methyl amine dehydrogenase, monoamine oxidase (MAO), diamine oxidase (DAO) etc., have been used. Among these, DAO is of particular interest as it shows high speci- ficity towards histamine and also it can oxidatively deaminate histamine [6]. To estimate histamine levels for fish freshness deter- mination various analytical methods such as high performance liquid chromatography [7,8], Spectrofluorimetry [9], thin layer chromatography [10], ELISA, capillary electrophoresis, impedance spectroscopy [11], microdialysis [12] etc., have been used. But all these methods require sample preparation and time consuming [13]. In a step of advancements in biosensors, Telsing et al. developed a voltammetric pea seedling amine oxidase modified biosensor which was able to detect cadaverine, putrescine with a linear range of 30–88 g mL -1 and 24–67 g mL -1 respectively, but it failed to detect histamine [14]. Lomillo et al. developed a disposable biosensor which showed poor precision in terms of repeata- bility [15]. Also, various electrochemical based biosensors were http://dx.doi.org/10.1016/j.snb.2014.04.009 0925-4005/© 2014 Published by Elsevier B.V.