Glutamate is a critical neurotransmitter in the central nervous system that plays a key role in numerous physiological processes and neurological disorders. Traditional methods of glutamate detection have low spatiotemporal resolution, while electrochemical methods are limited due to glutamate not being readily redox active at unmodified carbon electrode surfaces. This study presents the development of a glutamate oxidase-modified microelectrode for the sensitive, real-time detection of glutamate using fast-scan cyclic voltammetry (FSCV) with a triangle waveform. Here, we employed a chitosan-hydrogel coating to immobilize glutamate oxidase onto carbon fiber microelectrodes, enabling selective metabolism of glutamate to hydrogen peroxide. The metabolism to hydrogen peroxide facilitates indirect detection with high sensitivity across a concentration range relevant to physiological concentrations. We utilized FSCV for detection, which enhanced temporal resolution and chemical selectivity, allowing for the co-detection of glutamate with other neurotransmitters such as dopamine and norepinephrine. We performed proof-of-concept validation and testing utilizing both biological fluids and complex food samples, demonstrating the enzyme-modified microelectrode's broad applicability in clinical diagnostics and food quality assessment. The sensor showed excellent stability, resistance to fouling, and retained over 90% of its initial response after multiple uses. This work highlights the potential of this biosensor as a versatile tool for minimally invasive, biocompatible, rapid, and accurate glutamate measurement in a wide variety of samples for a diverse set of applications.

Voltammetric methods hold promise for the rapid and sensitive quantification of melatonin. This study reports the direct electrochemical quantification of melatonin using carbon nanotube (CNT) fiber cross-sections as microelectrodes. Six identical highly densified CNT fiber cross-sections were employed to quantify melatonin in the range of 0.05-100 μM. The limit of detection and quantification were 10 and 35 nM, respectively, with a sensitivity of 0.1322 nA/μM. Interference studies with uric acid, hypoxanthine, and ascorbic acid demonstrate its performance. Real-world application was highlighted by measuring melatonin in food, pharmaceutical, and human urine samples.

In developing countries, subsistence gold mining entails mixing metallic mercury with crushed sediments to extract gold. In this approach, the gold-mercury amalgam is heated to evaporate mercury and obtain gold. Thus, the highly volatile mercury can be absorbed through inhalation, resulting in adverse health effects. Urinalysis can be used to detect mercury, which is excreted in urine and feces, and correlate exposure with toxic effects. The current gold standard analytical methods are based on fluorescence or inductively coupled plasma mass spectrometry methods, but are expensive, time consuming, and are not easily accessible in countries where testing is needed. In this work, we report on a miniature electrochemical sensor that can rapidly detect mercury in urine at levels well below the US Biological Exposure Index (BEI) limit of 50 ppb (μg/L). The sensor is based on a thin-film gold electrode and anodic stripping voltammetry electroanalytical approach. The sensor successfully detected mercury at trace levels in urine, with a limit of detection of ~15 ppb Hg in the linear range of 20-80 ppb. With the low-cost disposable sensors and portable instrumentation, it is well suited for point-of-care applications.

A method for the determination of selected aromatic amino acid biomarkers of oxidative stress using microchip electrophoresis with electrochemical detection is described. The separation of the major reaction products of phenylalanine and tyrosine with reactive nitrogen and oxygen species was accomplished using ligand exchange micellar electrokinetic chromatography with a PDMS/glass hybrid chip. Electrochemical detection was achieved using a pyrolyzed photoresist film working electrode. The system was evaluated for the analysis of the products of the Fenton reaction with tyrosine and phenylalanine, and the reaction of peroxynitrite with tyrosine.

We report an in-plane extended nanopore Coulter counter (XnCC) chip fabricated in a thermoplastic via imprinting. The fabrication of the sensor utilized both photolithography and focused ion beam milling to make the microfluidic network and the in-plane pore sensor, respectively, in Si from which UV resin stamps were generated followed by thermal imprinting to produce the final device in the appropriate plastic (cyclic olefin polymer, COP). As an example of the utility of this in-plane extended nanopore sensor, we enumerated SARS-CoV-2 viral particles (VPs) affinity-selected from saliva and extracellular vesicles (EVs) affinity-selected from plasma samples secured from mouse models exposed to different ionizing radiation doses.

This work describes the sensitive voltammetric determination of favipiravir (FAV) based on its reduction for the first time with a low-cost and disposable pencil graphite electrode (PGE). In addition, the determination of FAV was also performed based on its oxidation. Differential pulse (DP) voltammograms recorded in 0.5 M HSO for the reduction of FAV show that peak currents increase linearly in the range of 1.0 to 600.0 μM with a limit of detection of 0.35 μM. The acceptable recovery values (98.9-106.0 %) obtained from a pharmaceutical tablet, real human urine, and artificial blood serum samples spiked with FAV confirm the high accuracy of the proposed method.

Here, a novel biosensing platform for the detection of SARS-CoV-2 usable both at voltammetric and impedimetric mode is reported. The platform was constructed on a multi-walled carbon nanotubes (MWCNTs) screen-printed electrode (SPE) functionalized by methylene blue (MB), antibodies against SARS-CoV-2 spike protein (SP), a bioactive layer of chitosan (CS) and protein A (PrA). The voltammetric sensor showed superior performances both in phosphate buffer solution (PBS) and spiked-saliva samples, with LOD values of 5.0±0.1 and 30±2.1 ng/mL, compared to 20±1.8 and 50±2.5 ng/mL for the impedimetric sensor. Moreover, the voltammetric immunosensor was tested in real saliva, showing promising results.

A sensitive detection of extremely toxic phenylpyrazole insecticide, 'Fipronil' is presented. Currently, the advancement of approaches for the detection of insecticides at low concentrations with less time is important for environmental safety assurance. Considering this fact, an effort has been made to develop an electrospun CoZnO nanofiber (NF) based label-free electrochemical system for the detection of fipronil. The CoZnO NF were characterized using different techniques including field emission scanning electron microscopy (FE-SEM), Energy Dispersive X-Ray Analysis (EDX), X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and Raman Spectroscopy. Based on the experimental results, the proposed platform displayed a linear response for fipronil in the attogram/mL range despite the multiple interfering agents. The sensitivity of the device was found to be 3.99 Kῼ (g/ml) cm. Limit of detection (LOD) and limit of quantification (LOQ) were calculated and found to be 112 ag mL and 340 ag mL respectively. Further, this proposed sensor will be implemented in the fields for the rapid and proficient detection of the real samples.

Contactless broadband microwave spectroscopy (a.k.a., broadband dielectric spectroscopy (BDS)) enables the accurate analysis of the electrical and magnetic properties without compromising the kinetic conditions of the experiment. The BDS method is sensitive to the actual electronic structure of species, and it is most relevant to redox reactions involving charge-transfer. In this paper, using BDS, we have studied and characterized the oxidation of a copper layer in a purposely built prototypical 3-D integrated circuit (3D-IC) during cycled high-temperature storage. We show that the microwave signal loss in these devices is attributable to the energy dissipation through the signal's interactions with the copper oxidation product. The results demonstrate that contactless BDS could be leveraged into an excellent metrology for applications that use metal oxide as sensing elements.

We describe a microfluidic device that can be used to detect interactions between red blood cells (RBCs) and endothelial cells using a gold pillar array (created by electrodeposition) and an integrated detection electrode. Endothelial cells can release nitric oxide (NO) via stimulation by RBC-derived ATP. These studies incorporate on-chip endothelial cell immobilization, direct RBC contact, and detection of NO in a single microfluidic device. In order to study the RBC-EC interactions, this work used a microfluidic device made of a PDMS chip with two adjacent channels and a polystyrene base with embedded electrodes for creating a membrane (via gold pillars) and detecting NO (at a glassy carbon electrode coated with platinum-black and Nafion). RBCs were pharmacologically treated with treprostinil in the absence and presence of glybenclamide, and ATP release was determined as was the resultant NO release from endothelial cells. Treprostinil treatment of RBCs resulted in ATP release that stimulated endothelial cells to release on average 1.8 ± 0.2 nM NO per endothelial cell (average ± SEM, = 8). Pretreatment of RBCs with glybenclamide inhibited treprostinil-induced ATP release and, therefore, less NO was produced by the endothelial cells (0.92 ± 0.1 nM NO per endothelial cell, = 7). In the future, this device can be used to study interactions between many other cell types (both adherent and non-adherent cell lines) and incorporate other detection schemes.

Microfluidic platforms can lead to miniaturisation, increased throughput and reduced reagent consumption, particularly when the processes are automated. Here, a programmable microcontroller is used for automation of a microfluidic platform configured to electrochemically determine the levels of 8 proteins simultaneously in complex liquid samples. The platform system is composed of a programmable Arduino microcontroller that controls inexpensive valve actuators, pump, magnetic stirrer and electronic display. The programmable microcontroller results in repeatable timing for each step in a complex assay protocol, such as sandwich immunoassays. Application of the platform is demonstrated using a multiplexed electrochemical immunoassay based on capture at the electrode surface of magnetic particles labelled with horseradish peroxidase and detection antibody. The multiplexed assay protocol is completed in less than 30 mins and results in detection of eight proteins associated with prostate cancer. The approach presented can be used to automate and simplify high-throughput screening campaigns, such as detection of multiple biomarkers in patient samples.

Capillary electrophoresis coupled with electrochemical detection can be a powerful analysis tool; however, previous methods developed to integrate these two techniques can often times be fragile and have alignment issues such that there are no commercially available approaches. In this paper, we present the use of a 3D-printed Wall-Jet Electrode device for integrating capillary electrophoresis with electrochemical detection. A pressure mobilization step was also utilized to further reduce noise by allowing the electrophoresis separation step to continue only until the first analyte was close to elution. Then, the separation voltage was terminated and pressure-based flow was used for elution of the analyte bands onto the electrode surface with a wall-jet configuration. It is shown that the pressure-based elution is beneficial for the reduction of baseline noise and elimination of field effects. A mixture of catecholamines were separated to demonstrate effectiveness of the system. In addition, the system was coupled with a Beckman Coulter commercial capillary electrophoresis instrument in a straightforward manner. The system was also shown to be effective in separations done with a high ionic strength physiological buffer. This 3D printing approach can be used by researchers to utilize electrochemical detection on commercial capillary electrophoresis systems by downloading the provided STL and/or CAD files.

Nitric oxide (NO) levels in exhaled breath are a non-invasive marker that can be used to diagnose various respiratory diseases and monitor a patient's response to given therapies. A portable and inexpensive device that can enable selective NO concentration measurements in exhaled breath samples is needed. Herein, the performance of an amperometric Pt-Nafion-based gas phase sensor for detection of NO in exhaled human nasal breath is examined. Enhanced selectivity over carbon monoxide and ammonia is achieved via an in-line zinc oxide-based filter. Exhaled nasal NO levels measured in 21 human samples with the sensor are shown to correlate well with those obtained using a chemiluminescence reference method (R = 0.9836).

Poly-L-lysine is one of the biocompatible polymers having amino and carboxyl groups in its structure. This attractive feature of poly-L-lysine makes it very convenient for bioactive molecule attachment. This study details the preparation of poly-L-lysine-based pencil graphite electrodes (PLL/PGEs) and use of the coated electrodes for direct ultrasensitive DNA hybridization detection. In the first part of this study, poly-L-lysine coated electrodes were prepared using L-lysine as the monomer by cyclic voltammetry (CV) with different cyclic scans. The effect of these cyclic scans during the electropolymerization was investigated. Coated electrodes were characterized by cyclic voltammetry, electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). Then, one-pot preparation of poly-L-lysine composites with graphene (GN) and multi-walled carbon nanotubes (MWCNTs) onto the pencil graphite electrodes were achieved. Electrochemical responses of these 3 electrodes were compared. After all, electrochemical DNA hybridization was performed using the poly-L-lysine-based electrodes prepared at optimum polymerization condition. The PLL/PGE coated electrode presented a good linear response in the target concentration range of 1.0×10 to 1.0×10 with a detection limit of 2.25×10 using differential pulse voltammetry as the detection method. We believe that poly-L-lysine-based surfaces will be useful for further clinical applications.

Huntington's disease (HD) is a fatal neurodegenerative disorder that is characterized by degeneration of the striatum. Here, fast-scan cyclic voltammetry at carbon-fiber microelectrodes was used to uncover regional differences in dopamine (DA) release in the caudate putamen of R6/2 and wild-type control mice. We found a decreasing ventral-to-dorsal gradient in DA release, evoked by a single electrical stimulus pulse, in aged R6/2 mice. Moreover, under more intense stimulation conditions (120 pulses), DA release was significantly attenuated in the dorsal, but not in the ventral caudate. Autoradiography measurements using [H]WIN 35,428 revealed that the overall density of DA transporter (DAT) protein molecules was significantly less in R6/2 mice compared to WT control mice; however, quadrants of the caudate putamen were not differentially altered in the R6/2 mice. These data collectively suggest that DA release in the dorsal caudate region is more vulnerable with age progression compared to the ventral region.

Carbon nanohorns (CNHs), closed cone-shaped cages of -hybridized carbons, are a promising nanomaterial to improve carbon-fiber microelectrode (CFME) dues to their high specific surface area and edge planes, but few studies have tested their electrochemical properties. Here, we tested the dopamine detection at electrodeposited CNHs on CFME (CNH/CFME). The optimized concentration of CNHs in the deposition solution is 0.5 mg/mL, and the optimized electrodeposition waveform is 10 cycles of triangular waveform scanned from -1.0 V and +1.0 V at 50 mV/s. Using fast-scan cyclic voltammetry, the optimized CNH/CFME enhances dopamine peak current to 2.3 ± 0.2 times that of the CFME. To further increase the current, CNH/CFMEs were oxidized in NaOH (ox-CNH/CFME), which creates more defects and surface oxide groups to adsorb dopamine. The oxidative etching further increases the peak current to 3.5 ± 0.2 times of the CFME, and ox-CNH/CFME had a limit of detection of 6 ± 2 nM. The dopamine anodic current at ox-CNH/CFME was stable for 8 h of continuous scanning. The ox-CNH/CFME enhanced the anodic peak current for other cationic neurotransmitters including epinephrine, norepinephrine, and serotonin, but less enhancement was found for ascorbic acid, showing higher selectivity for cationic molecules. CNHs also decreased tissue biofouling at CFME. Thus, electrodeposited CNHs are a promising new method for increasing the surface area and current of CFMEs for dopamine detection.

High-performance biosensors were fabricated by efficiently transferring enzyme onto Pt electrode surfaces using a polydimethylsiloxane (PDMS) stamp. Polypyrrole and Nafion were coated first on the electrode surface to act as permselective films for exclusion of both anionic and cationic electrooxidizable interfering compounds. A chitosan film then was electrochemically deposited to serve as an adhesive layer for enzyme immobilization. Glucose oxidase (GOx) was selected as a model enzyme for construction of a glucose biosensor, and a mixture of GOx and bovine serum albumin was stamped onto the chitosan-coated surface and subsequently crosslinked using glutaraldehyde vapor. For the optimized fabrication process, the biosensor exhibited excellent performance characteristics including a linear range up to 2 mM with sensitivity of 29.4 ± 1.3 μA mM cm and detection limit of 4.3 ± 1.7 μM (S/N = 3) as well as a rapid response time of ~2 s. In comparison to those previously described, this glucose biosensor exhibits an excellent combination of high sensitivity, low detection limit, rapid response time, and good selectivity. Thus, these results support the use of PDMS stamping as an effective enzyme deposition method for electroenzymatic biosensor fabrication, which may prove especially useful for the deposition of enzyme at selected sites on microelectrode array microprobes of the kind used for neuroscience research .

In this work, we report on the determination of trace manganese (Mn) using cathodic stripping voltammetry (CSV) using a microfabricated sensor with a Pt thin-film working electrode. While an essential trace metal for human health, prolonged exposure to Mn tends to gradually impair our neurological system. The potential sources of Mn exposure make it necessary to monitor the concentration in various sample matrices. Previous work by us and others suggested CSV as an effective method for measuring trace Mn. The analytical performance metrics were characterized and optimized, leading to a calculated limit of detection (LOD) of 16.3 nM (0.9 ppb) in pH 5.5, 0.2 M acetate buffer. Further, we successfully validated Mn determination in surface water with ~90% accuracy and >97% precision as compared with ICP-MS "gold standard" measurement. Ultimately, with stable, accurate and precise electrochemical performance, this Pt sensor permits rapid monitoring of Mn in environmental samples, and could potentially be used for point-of-use measurements if coupled with portable instrumentation.

Electrochemical hybridization sensors have been explored extensively for analysis of specific nucleic acids. However, commercialization of the platform is hindered by the need for attachment of separate oligonucleotide probes complementary to a RNA or DNA target to an electrode's surface. Here we demonstrate that a single probe can be used to analyze several nucleic acid targets with high selectivity and low cost. The universal electrochemical four-way junction (4J)-forming (UE4J) sensor consists of a universal DNA stem-loop (USL) probe attached to the electrode's surface and two adaptor strands (m and f) which hybridize to the USL probe and the analyte to form a 4J associate. The m adaptor strand was conjugated with a methylene blue redox marker for signal ON sensing and monitored using square wave voltammetry. We demonstrated that a single sensor can be used for detection of several different DNA/RNA sequences and can be regenerated in 30 seconds by a simple water rinse. The UE4J sensor enables a high selectivity by recognition of a single base substitution, even at room temperature. The UE4J sensor opens a venue for a re-useable universal platform that can be adopted at low cost for the analysis of DNA or RNA targets.

An array of four independently wired indium tin oxide (ITO) electrodes was used for electrochemically stimulated DNA release and activation of DNA-based Identity, AND and XOR logic gates. Single-stranded DNA molecules were loaded on the mixed poly(,-di-methylaminoethyl methacrylate) (PDMAEMA)/poly-(methacrylic acid) (PMAA) brush covalently attached to the ITO electrodes. The DNA deposition was performed at pH 5.0 when the polymer brush is positively charged due to protonation of tertiary amino groups in PDMAE-MA, thus resulting in electrostatic attraction of the negatively charged DNA. By applying electrolysis at -1.0 V(vs. Ag/AgCl reference) electrochemical oxygen reduction resulted in the consumption of hydrogen ions and local pH increase near the electrode surface. The process resulted in recharging the polymer brush to the negative state due to dissociation of carboxylic groups of PMAA, thus repulsing the negatively charged DNA and releasing it from the electrode surface. The DNA release was performed in various combinations from different electrodes in the array assembly. The released DNA operated as input signals for activation of the Boolean logic gates. The developed system represents a step forward in DNA computing, combining for the first time DNA chemical processes with electronic input signals.

Acetaminophen (APAP) is an antipyretic, analgesic agent, the overdose of which during medical treatment poses a risk for liver failure. Hence, it is important to develop methods to monitor physiological APAP levels to avoid APAP. Here, we report an efficient, selective electrochemical APAP sensor made from depositing silicon nanowires (SiNWs) onto glassy carbon electrodes (GCEs). Electrocatalytic activity of the SiNW/GCE sensors was monitored under varying pH and concentrations of APAP using cyclic voltammetry (CV) and chronoamperometry (CA). CV of the SiNWs at 0.5 to 13 mmol dm APAP concentrations was used to determine the oxidation and reduction potential of APAP. The selective detection of APAP was then demonstrated using CA at +0.568 V vs Ag/AgCl, where APAP is fully oxidized, in the 0.01 to 3 mmol dm concentration range with potentially-interfering species. The SiNW sensor has the ability to detect APAP well within the detection limits for APAP toxicity, showing promise as a practical biosensor.