A Nafion/AChE-cSWCNT/MWCNT/Au-based amperometric biosensor for the determination of organophosphorous compounds.

In the present study, a biosensor was developed for the detection of organophosphorous compounds. Core electrode of a working electrode was obtained by depositing the paste of Gold nanoparticles and Multi-walled Carbon Nanotubes on a gold wire. The acetylcholinesterase enzyme was immobilized on carboxylated Single-walled Carbon Nanotubes and pasted onto a core of electrode followed by coating with a nafion layer to prevent enzyme leaching from the electrode. This electrode was further used as a working electrode in the sensor. This sensor worked on the AChE inhibition mechanism where the signal is inversely proportional to the amount of organophosphorous compounds. The electrocatalytic activity of this sensor was observed at a potential of +0.360 mV. The standardized conditions for this sensor were pH at 7.0, temperature at 30°C and response time at less than 10s. The linear working range of this biosensor was 0.1-130 µM with the lowest detection limit (LOD) of 1.9, 2.3, 2.2 and 2.5 nM for Methyl Parathion, Monocrotophos, Chlorpyrifos and Endosulfan, respectively. The biosensor showed excellent reusability (upto 55 times) and can be stored stably for 2 months.

Fabrication of AChE/SnO2-cMWCNTs/Cu Nanocomposite-Based Sensor Electrode for Detection of Methyl Parathion in Water.

The work highlights inhibition-based Acetylcholinesterase (AChE) fabrication using composite nanomaterial comprising tin oxide nanoparticles (SnO2) and carboxylated multiwalled carbon nanotubes (cMWCNTs) for detection of pesticide methyl parathion (MP) in water samples. Working electrode AChE/SnO2-cMWCNTs/Cu exhibited high sensitivity with a linearity range of 1.0 µM to 160 µM and a minimum detection limit of 0.1 µM for MP in water. The fabricated electrode was found biocompatible and nontoxic which can be used to detect low concentrations of pesticide in water samples. The synergistic and facilitated electron transferring properties of SnO2-cMWCNTs/Cu made it an excellent support for immobilization of enzyme in sensing technology. The enzyme AChE was covalently immobilized with cMWCNTs using glutaraldehyde as crosslinking agent which has enhanced the storage stability and reusability of the method. The reusability attained was 30 times for 40 days when AChE/SnO2-cMWCNTs/Cu was stored at low temperature of 4°C. Developed sensor showed excellent analytical recovery of pesticide in water sample with negligible effect of interfering species. Also, AChE/SnO2-cMWCNTs/Cu was easily reactivated simply by varying pH of phosphate buffer. This method is fast, reliable, and accurate showing successful development of amperometric biosensor for detection of MP in water sample.

Recent approaches to ameliorate selectivity and sensitivity of enzyme based cholesterol biosensors: a review.

The healthcare area is often reluctant to execute new technology unless they are proven to be safe, constructive and secure. Eventually, an aspiration stands for providing point-of-care testing service to allow a better estimation of the biochemical levels of a patient that entails an insistent remedial action. With increasing mortality rate due to cardiovascular diseases (CVDs) in present scenario, it has become the need of hour to develop more advance methods for their diagnosis, so that it can be determined at sensitive levels and can be prevented from being fatal. Elevated level of cholesterol in blood stream is one of the utmost risk factors which lead to CVDs. Discernible from the vast research in this field, worth of cholesterol biosensors is already recognized and flourished in the clinical analysis of brain and cardiac vascular diseases. It necessitates unremitting progress in the development of biosensing technology towards fabrication, miniaturization and multiplexing ability of cholesterol quantification devices so that they can endow with lab-on-chip-analysis systems to the medical field. Different strategies have been meticulously explored for the engineering of cholesterol biosensors utilizing nanocomposites, conducting polymers, nanotubes and nanoparticles. Foremost, this article reviews the contemporary evolution in cholesterol biosensors, which encompass various strategies for immobilization of enzymes and roles of various matrices and artificial mediators used for the biosensor fabrication. Still there remains an enormous challenge to congregate the demands of performance and yield in a cost effective manner for its application in successful treatments of CVDs.

Fabrication and Optimization of ChE/ChO/HRP-AuNPs/c-MWCNTs Based Silver Electrode for Determining Total Cholesterol in Serum.

The developed method used three enzymes comprised of cholesterol esterase, cholesterol oxidase, and peroxidase for fabrication of amperometric biosensor in order to determine total cholesterol in serum samples. Gold nanoparticles (AuNPs) and carboxylated multiwall carbon nanotubes (cMWCNTs) were used to design core of working electrode, having covalently immobilized ChO, ChE, and HRP. Polyacrylamide layer was finally coated on working electrode in order to prevent enzyme leaching. Chemically synthesised Au nanoparticles were subjected to transmission electron microscopy (TEM) for analysing the shape and size of the particles. Working electrode was subjected to FTIR and XRD. The combined action of AuNP and c-MWCNT showed enhancement in electrocatalytic activity at a very low potential of 0.27 V. The pH 7, temperature 40°C, and response time of 20 seconds, respectively, were observed. The biosensor shows a broad linear range from 0.5 mg/dL to 250 mg/dL (0.01 mM-5.83 mM) with minimum detection limit being 0.5 mg/dL (0.01 mM). The biosensor showed reusability of more than 45 times and was stable for 60 days. The biosensor was successfully tested for determining total cholesterol in serum samples amperometrically with no significant interference by serum components.

Biosensors based on electrochemical lactate detection: A comprehensive review.

Lactate detection plays a significant role in healthcare, food industries and is specially necessitated in conditions like hemorrhage, respiratory failure, hepatic disease, sepsis and tissue hypoxia. Conventional methods for lactate determination are not accurate and fast so this accelerated the need of sensitive biosensors for high-throughput screening of lactate in different samples. This review focuses on applications and developments of various electrochemical biosensors based on lactate detection as lactate being essential metabolite in anaerobic metabolic pathway. A comparative study to summarize the L-lactate biosensors on the basis of different analytical properties in terms of fabrication, sensitivity, detection limit, linearity, response time and storage stability has been done. It also addresses the merits and demerits of current enzyme based lactate biosensors. Lactate biosensors are of two main types - lactate oxidase (LOD) and lactate dehydrogenase (LDH) based. Different supports tried for manufacturing lactate biosensors include membranes, polymeric matrices-conducting or non-conducting, transparent gel matrix, hydrogel supports, screen printed electrodes and nanoparticles. All the examples in these support categories have been aptly discussed. Finally this review encompasses the conclusion and future emerging prospects of lactate sensors.

Preparation of electrochemical biosensor for detection of organophosphorus pesticides.

Polyvinyl chloride (PVC) can be used to develop reaction beaker which acts as electrochemical cell for the measurement of OP pesticides. Being chemically inert, corrosion resistant, and easy in molding to various shapes and size, PVC can be used for the immobilization of enzyme. Organophosphorus hydrolase was immobilized covalently onto the chemically activated inner surface of PVC beaker by using glutaraldehyde as a coupling agent. The carbon nanotubes paste working electrode was constructed for amperometric measurement at a potential of +0.8 V. The biosensor showed optimum response at pH 8.0 with incubation temperature of 40°C. K m and I max for substrate (methyl parathion) were 322.58 µM and 1.1 µA, respectively. Evaluation study showed a correlation of 0.985, which was in agreement with the standard method. The OPH biosensor lost 50% of its initial activity after its regular use for 25 times over a period of 50 days when stored in 0.1 M sodium phosphate buffer, pH 8.0 at 4°C. No interference was observed by interfering species.

Acetylcholinesterase biosensors for electrochemical detection of organophosphorus compounds: a review.

The exponentially growing population, with limited resources, has exerted an intense pressure on the agriculture sector. In order to achieve high productivity the use of pesticide has increased up to many folds. These pesticides contain organophosphorus (OP) toxic compounds which interfere with the proper functioning of enzyme acetylcholinesterase (AChE) and finally affect the central nervous system (CNS). So, there is a need for routine, continuous, on spot detection of OP compounds which are the main limitations associated with conventional analytical methods. AChE based enzymatic biosensors have been reported by researchers as the most promising tool for analysis of pesticide level to control toxicity and for environment conservation. The present review summarises AChE based biosensors by discussing their characteristic features in terms of fabrication, detection limit, linearity range, time of incubation, and storage stability. Use of nanoparticles in recently reported fabrication strategies has improved the efficiency of biosensors to a great extent making them more reliable and robust.