Abstract
This work describes the investigation of a label-free immunosensor for the detection of aflatoxin B1 (AFB1). CD-trodes (electrodes obtained from recordable compact disks) were used as low-cost and disposable transducers after modification with a self-assembled monolayer (SAM) of lipoic acid. The anti-aflatoxin B1 antibody was immobilized via EDC/NHS activation, followed by blocking with bovine serum albumin and immunoassays with AFB1. The optimization of analytical parameters and the detection were carried out using electrochemical impedance measurements. Using chemometric tools, the best conditions for the immunosensor development were defined as: anti-AFB1 antibody at 1:2000 dilution and surface blocking with 0.5% bovine serum albumin, both incubated for 1 h, and antibody–antigen immunoreaction for 30 min. The impedimetric immunosensor showed a linear range from 5 × 10−9 to 1 × 10−7 mol·L−1 (1.56–31.2 ng·mL−1), limit of detection and limit of quantification, respectively, 3.6 × 10−10 and 1.1 × 10−9mol·L−1 (0.11 and 0.34 ng·mL−1). The proposed immunosensor was applied to analyze peanut samples.
Highlights
Aflatoxin B1 (AFB1 ) is a toxic metabolite produced by fungi Aspergillus flavus and A. parasiticus that is characterized by acute toxicity, teratogenicity, mutagenicity, and carcinogenicity [1]
This work reports the development of an impedimetric immunosensor for the determination of aflatoxin B1, through gold CD-trode surface modification with a self-assembled monolayer (SAM) of lipoic acid activated via EDC (1-ethyl-3-(3-(dimethylamino)-propyl)carbodiimide)/NHS (N-hydroxy succinimide) for the immobilization of anti-aflatoxin B1 antibody
EDC/NHS prepared in deionized water that was added on the surface of the modified electrode, and the incubation time was 60 min (Scheme 1-2) [32]
Summary
Aflatoxin B1 (AFB1 ) is a toxic metabolite produced by fungi Aspergillus flavus and A. parasiticus that is characterized by acute toxicity, teratogenicity, mutagenicity, and carcinogenicity [1]. The acute effects are observed mainly in the liver; they can result in necrosis, hemorrhage, injury, fibrosis, cirrhosis, and cancer This toxin infects a wide range of agricultural products [2], especially peanuts, corn, wheat, rice, nuts, dried fruits, among others. An alternative is offered by the use of immunosensors, due to their sensibility, stability, and ease of handling In this case, the antibodies are immobilized on an electrode and must maintain their biological activity on the transducer [8]. This work reports the development of an impedimetric immunosensor for the determination of aflatoxin B1 , through gold CD-trode (electrode obtained from recordable compact disks) surface modification with a self-assembled monolayer (SAM) of lipoic acid activated via EDC (1-ethyl-3-(3-(dimethylamino)-propyl)carbodiimide)/NHS (N-hydroxy succinimide) for the immobilization of anti-aflatoxin B1 antibody. The proposed biosensor was applied to determine the antigen in peanut samples
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