A label-free electrochemical impedance immunosensor for the sensitive detection of aflatoxin B1

Because of the potential health impact of aflatoxin B1 (AFB1), it is essential to monitor the level of this mycotoxin in a variety of foods and agricultural products. In this paper, a novel immunosensor for the rapid detection of AFB1 based on label-free electrochemical impedance spectroscopy (EIS) monitoring was achieved. The immunosensor was fabricated by stepwise immobilization of 1,6-hexanedithiol, colloidal Au, and aflatoxin B1–bovine serum albumin conjugate (AFB1–BSA) on a gold electrode via self-assembling technique. The interfacial properties of the modified electrodes were evaluated using the Fe(CN)63−/4− redox couple as a probe via cyclic voltammetry (CV) and EIS. An equivalent circuit model with a constant phase element was used to interpret the obtained impedance spectra. The impedance via the specific immuno-interaction at the sensor surface was utilized to detect AFB1 in samples. Under the optimized conditions, the impedance increment was linearly related to the AFB1 concentration in the range of 0.08 to 100 ng mL−1 with a detection limit of 0.05 ng mL−1 (S/N = 3) and a correlation coefficient of 0.9919.

[1]  J. Randles Kinetics of rapid electrode reactions , 1947 .

[2]  Guo-Li Shen,et al.  Impedance immunosensor based on receptor protein adsorbed directly on porous gold film , 2005 .

[3]  Mohammad Sarwar Nasir,et al.  Development of a fluorescence polarization assay for the determination of aflatoxins in grains. , 2002, Journal of agricultural and food chemistry.

[4]  Jean-Louis Marty,et al.  Label-free impedimetric immunosensor for sensitive detection of ochratoxin A. , 2009, Biosensors & bioelectronics.

[5]  Jian-Ding Qiu,et al.  A label-free amperometric immunosensor based on biocompatible conductive redox chitosan-ferrocene/gold nanoparticles matrix. , 2009, Biosensors & bioelectronics.

[6]  Guo-Li Shen,et al.  Reusable electrochemical sensing platform for highly sensitive detection of small molecules based on structure-switching signaling aptamers. , 2007, Analytical chemistry.

[7]  Xiulan Sun,et al.  A Simple, Highly Sensitive, and Label-Free Impedimetric Immunosensor for Detection of Microcystin-LR in Water , 2010 .

[8]  C. Maragos,et al.  Analysis of Aflatoxin B1 in Corn Using Capillary Electrophoresis with Laser-Induced Fluorescence Detection , 1997 .

[9]  M. Agut,et al.  Determination of aflatoxins B1, G1, B2 and G2 in medicinal herbs by liquid chromatography-tandem mass spectrometry. , 2004, Journal of chromatography. A.

[10]  C. Cavaliere,et al.  Determination of aflatoxins in olive oil by liquid chromatography-tandem mass spectrometry. , 2007, Analytica chimica acta.

[11]  Song Zhang,et al.  A sensitive impedance immunosensor based on functionalized gold nanoparticle-protein composite films for probing apolipoprotein A-I. , 2007, Talanta.

[12]  T. Yoshizawa,et al.  In-house direct cELISA for determining aflatoxin B 1 in Thai corn and peanuts , 2003, Food additives and contaminants.

[13]  H. Karnes,et al.  Determination of aflatoxin B1 in sidestream cigarette smoke by immunoaffinity column extraction coupled with liquid chromatography/mass spectrometry. , 2005, Journal of chromatography. A.

[14]  E Anklam,et al.  Validation of analytical methods for determining mycotoxins in foodstuffs , 2002 .

[15]  Itamar Willner,et al.  Label-free and reagentless aptamer-based sensors for small molecules. , 2006, Journal of the American Chemical Society.

[16]  Zhao-xiang Zhang,et al.  Determination of aflatoxins in high-pigment content samples by matrix solid-phase dispersion and high-performance liquid chromatography. , 2006, Journal of agricultural and food chemistry.

[17]  Frank Davis,et al.  Label-free and reversible immunosensor based upon an ac impedance interrogation protocol , 2005 .

[18]  Shi Li,et al.  Amperometric biosensor for aflatoxin B1 based on aflatoxin-oxidase immobilized on multiwalled carbon nanotubes , 2011 .

[19]  G. Shen,et al.  A signal-amplified electrochemical immunosensor for aflatoxin B(1) determination in rice. , 2009, Analytical biochemistry.

[20]  Yanbin Li,et al.  Interdigitated Array microelectrode-based electrochemical impedance immunosensor for detection of Escherichia coli O157:H7. , 2004, Analytical chemistry.

[21]  R. G. Freeman,et al.  Preparation and Characterization of Au Colloid Monolayers , 1995 .

[22]  H. Reinhard,et al.  Reversed-phase liquid chromatographic behavior of the mycotoxins citrinin and ochratoxin A. , 1999, Journal of chromatography. A.

[23]  Joseph Wang,et al.  Label-free bioelectronic detection of aptamer–protein interactions , 2005 .

[24]  Joseph Owino,et al.  Electrochemical Immunosensor Based on Polythionine/Gold Nanoparticles for the Determination of Aflatoxin B1 , 2008, Sensors.

[25]  G. Bang,et al.  A novel electrochemical detection method for aptamer biosensors. , 2005, Biosensors & bioelectronics.

[26]  B. Kamp,et al.  Development of an electrochemical immunosensor for direct detection of interferon-γ at the attomolar level , 2001 .

[27]  J. Chen,et al.  Thin-layer chromatography of mycotoxins and comparison with other chromatographic methods. , 1998, Journal of chromatography. A.

[28]  S. Hajare,et al.  Development of a radioimmunoassay procedure for aflatoxin B1 measurement. , 2003, Journal of agricultural and food chemistry.

[29]  J Gilbert,et al.  Overview of mycotoxin methods, present status and future needs. , 1999, Natural toxins.