P. Raghu

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

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 0.1M 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.

Acetylcholinesterase based biosensor for monitoring of Malathion and Acephate in food samples: A voltammetric study

Acetylcholinesterase (AChE) biosensor was developed through silica sol-gel (SiSG) immobilisation of AChE on the carbon paste electrode (CPE) and used as working electrode. AChE catalyses the cleavage of acetylthiocholine chloride (ASChCl or substrate) to thiocholine, which was oxidised to give a disulphide compound by dimerisation at 0.60V versus saturated calomel electrode. All the experiments were carried out in 0.1M phosphate buffer solution (PBS) at pH 7.0 and 0.1M KCl solution at room temperature. The limit of detection and limit of quantification values were found to be 0.058ppm, 0.044ppm and 0.194ppm, 0.147ppm for Malathion and Acephate, respectively. The response of the biosensor showed a good linearity range with an incubation time of 4min for Malathion and Acephate, respectively. This biosensor was used for the direct determination of pesticides without any pretreatment and it requires less time for analysis.

A novel electrochemical biosensor based on horseradish peroxidase immobilized on Ag-nanoparticles/poly(l-arginine) modified carbon paste electrode toward the determination of pyrogallol/hydroquinone

A novel electrochemical biosensor for the determination of pyrogallol (PG) and hydroquinone (HQ) has been constructed based on the poly l-arginine (poly(l-Arg))/carbon paste electrode (CPE) immobilized with horseradish peroxidase (HRP) and silver nanoparticles (AgNPs) through the silica sol-gel (SiSG) entrapment. The electrochemical properties of the biosensor were characterized by employing the electrochemical techniques. The proposed biosensor showed a high sensitivity and fast response toward the determination of PG and HQ around 0.18V. Under the optimized conditions, the anodic peak current of PG and HQ was linear with the concentration range of 8μM to 30×10(-5)M and 1-150μM. The limit of detection (LOD) and limit of quantification (LOQ) were found to be 6.2μM, 20μM for PG and 0.57μM, 1.92μM for HQ respectively. The electrochemical impedance spectroscopy (EIS) studies have confirmed that the occurrence of electron transfer at HRP-SiSG/AgNPs/poly(l-Arg)/CPE was faster. Moreover the stability, reproducibility and repeatability of the biosensor were also studied. The proposed biosensor was successfully applied for the determination of PG and HQ in real samples and the results were found to be satisfactory.

A novel horseradish peroxidase biosensor towards the detection of dopamine: a voltammetric study.

A polymerized film of glycine (Gly) was prepared on the surface of carbon paste electrode (CPE) through the cyclic voltammetry (CV) technique. A novel biosensor for the determination of dopamine (DA) has been constructed based on horseradish peroxidase (HRP) and multiwalled carbon nanotubes (MWCNTs) immobilizing on Poly (Gly)/CPE through silica sol-gel (SiSG) entrapment. CV measurements were employed in order to understand the feasibility of poly (Gly) as an electron carrier between the immobilized peroxidase and the surface of CPE. By using differential pulse voltammetry (DPV) the calibration curves of DA was obtained in the range of 15-865 μM. The limit of detection (LOD) and limit of quantification (LOQ) of DA was found to be 6×10⁻⁷ M and 2×10⁻⁶ M respectively. The apparent Michaelis-Menten constant (Km(app)) was found to be 0.5 mM and illustrated that the good biological activity of the fixed enzyme. Electrochemical impedance spectroscopy (EIS) results confirmed the rapid electron transfer and also the immobilization of enzyme on the electrode surface. The biosensor showed high sensitivity, selectivity and reproducibility. This method has been used to determine DA in the presence of various interferences and in clinical preparations.

An electrochemical sensor based on poly (solochrome dark blue) film coated electrode for the determination of dopamine and simultaneous separation in the presence of uric acid and ascorbic acid: a voltammetric method.

In this paper, the carbon paste electrode (CPE) was modified with poly- Solochrome dark blue (poly-SDB), through polymerization process, and it was used for the sensitive and selective determination of dopamine (DA). The simultaneous determination of Dopamine (DA) Ascorbic acid (AA) and Uric acid (UA) in 0.1 M Phosphate buffer solution (PBS) of pH 6.0 was carried out by employing differential pulse voltammetric technique (DPV). The effect of pH, concentration, scan rate, accumulation time, repetability and reproducibility of DA was studied at poly-SDB film modified electrode. The poly-SDBCPE exhibited a strong electrocatalytic action toward the oxidation of DA, AA and UA. The overlapping voltammetric response of biomolecules separated in to three well defined peaks with lower oxidation potential and significant increase in the anodic peak currents in the presence of poly-SDBCPE was achieved. The results showed a good sensitivity, selectivity and high reproducibility of electro synthesized polymer electrode. The limit of detection, limit of quantification and correlation coefficient of DA at poly-SDBCPE was 0.8 μM, 2.8 μM and 0.99765 respectively. The effect of interference was studied by DPV technique. The developed modified electrode was used for the analysis of DA in pharmaceutical formulations with satisfactory results. The interfacial electron transfer behavior of DA was studied by electrochemical impedance spectroscopy (EIS) and results showed that the charge transfer rate was enhanced at poly- SDBCPE, when compared with bare CPE, glassy carbon electrode (GCE) and platinum electrode (Pt).