Sol–gel immobilized biosensor for the detection of organophosphorous pesticides: A voltammetric method

• Published in: Bioelectrochemistry (Amsterdam, Netherlands) Netherlands), 2012-02, Vol.83, p.19-24
• Main Authors: Raghu, P., Kumara Swamy, B.E., Madhusudana Reddy, T., Chandrashekar, B.N., Reddaiah, K.
• Format: Article
• Language: English
• Subjects: Acetylcholinesterase; Acetylcholinesterase - chemistry; Acetylcholinesterase - metabolism; Acetylthiocholine - metabolism; Acetylthiocholine chloride; Animals; Biosensing Techniques - methods; Carbon paste electrode; Electrochemistry - methods; Electrodes; Electrophorus; Enzymes, Immobilized - chemistry; Enzymes, Immobilized - metabolism; Graphite - chemistry; Hydrogen-Ion Concentration; Immobilization; Limit of Detection; Methyl Parathion - analysis; Methyl parathion and acephate; Ointments - chemistry; Organothiophosphorus Compounds - analysis; Oxidation-Reduction; Pesticides - analysis; Phase Transition; Phosphoramides; Potentiometry - methods; Silica Gel - chemistry
• ISSN: 1567-5394; 1878-562X
• DOI: 10.1016/j.bioelechem.2011.08.002

Organophosphorous compounds are important neuroactive molecules whose presence exhibits significant analytical challenges. An acetylcholinesterase (AChE) based amperometric biosensor was developed by silica sol–gel film immobilization of the enzyme onto the carbon paste electrode. The mono enzyme biosensor was used for the determination of two organophosphorous compounds such as methyl parathion (MP) and acephate in 0.1 M phosphate buffer (pH 7.0). The substrate used was acetylthiocholine chloride (ASChCl) confirmed the formation of thiocholine and it was electrochemically oxidized giving significant increase in anodic peak current around at 0.60 V versus calomel electrode. The influence of pH, enzyme loading and substrate concentration on the response of the biosensor was investigated. The monoenzyme biosensor provided linearity to methyl parathion and acephate in the concentration range of 0.1–0.5 ppb and 50–750 ppb with an incubation time of 20 min and 4 min. The detection limits under the optimum working conditions were found to be 0.08 ppb for methyl parathion and 87 ppb for acephate. The sensor shows good operational stability 89% of its original activity for 60 successive measurements. ► This paper describes the determination of organophosphorous pesticides. ► Biosensor, prepared through sol–gel immobilization method for the AChE entrapment. ► LOD of the biosensor was related to the capacity of the pesticide to inhibit AChE. ► Fast response, good reproducibility and sensitive enough to determine pesticides.