Innovative Graphene-Based Nanocomposites for Improvement of Electrochemical Sensors: Synthesis, Characterization, and Applications.

Graphene, renowned for its exceptional physicochemical attributes, has emerged as a favored substrate for integrating a wide array of inorganic and organic materials in scientific endeavors and innovations. Electrochemical graphene-based nanocomposite sensors have been developed by incorporating diverse nanoparticles into graphene, effectively immobilized onto electrodes through various techniques. These graphene-based nanocomposite sensors have effectively detected and quantified various electroactive species in samples. This review delves into using graphene nanocomposites to fabricate electrochemical sensors, leveraging the exceptional electrical, mechanical, and thermal properties inherent to graphene derivatives. These nanocomposites showcase electrocatalytic activity, substantial surface area, superior electrical conductivity, adsorption capabilities, and notable porosity, which are highly advantageous for sensing applications. A myriad of characterization techniques, including Raman spectroscopy, scanning electron microscopy (SEM), transmission electron microscopy (TEM), BET surface area analysis, and X-ray diffraction (XRD), have proven effective in exploring the properties of graphene nanocomposites and validating the adjustable formation of these nanomaterials with graphene. The applicability of these sensors across various matrices, encompassing environmental, food, and biological domains, has been evaluated through electrochemical measurements, such as cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS), and differential pulse voltammetry (DPV). This review provides a comprehensive overview of synthesis methods, characterization techniques, and sensor applications pertinent to graphene-based nanocomposites. Furthermore, it deliberates on the challenges and future prospects within this burgeoning field.

Patulin-imprinted origami 3D-ePAD based on graphene screen-printed electrode modified with Mn-ZnS quantum dot coated with a molecularly imprinted polymer.

We developed a novel, compact, three-dimensional electrochemical paper-based analytical device (3D-ePAD) for patulin (PT) determination. The selective and sensitive PT-imprinted Origami 3D-ePAD was constructed based on a graphene screen-printed electrode modified with manganese-zinc sulfide quantum dots coated with patulin imprinted polymer (Mn-ZnS QDs@PT-MIP/GSPE). The Mn-ZnS QDs@PT-MIP was synthesized using 2-oxindole as the template, methacrylic acid (MAA) as a monomer, N,N'-(1,2-dihydroxyethylene) bis (acrylamide) (DHEBA) as cross-linker and 2,2'-azobis (2-methylpropionitrile) (AIBN) as initiator, respectively. The Origami 3D-ePAD was designed with hydrophobic barrier layers formed on filter paper to provide three-dimensional circular reservoirs and assembled electrodes. The synthesized Mn-ZnS QDs@PT-MIP was quickly loaded on the electrode surface by mixing with graphene ink and then screen-printing on the paper. The PT-imprinted sensor provides the greatest enhancement in redox response and electrocatalytic activity, which we attributed to synergetic effects. This arose from an excellent electrocatalytic activity and good electrical conductivity of Mn-ZnS QDs@PT-MIP, which improved electron transfer between PT and the electrode surface. Under the optimized DPV conditions, a well-defined PT oxidation peak appears at +0.15 V (vs Ag/AgCl) using 0.1 M of phosphate buffer (pH 6.5) containing 5 mM K3Fe(CN)6 as the supporting electrolyte. Our developed PT imprinted Origami 3D-ePAD revealed excellent linear dynamic ranges of 0.001-25 µM, with a detection limit of 0.2 nM. Detection performance indicated that our Origami 3D-ePAD possesses outstanding detection performance from fruits and CRM in terms of high accuracy (%Error for inter-day is 1.11%) and precision (%RSD less than 4.1%). Therefore, the proposed method is well-suited as an alternative platform for ready-to-use sensors in food safety. The imprinted Origami 3D-ePAD is an excellent disposable device with a simple, cost-effective, and fast analysis, and it is ready to use for determining patulin in actual samples.

An alternative ready-to-use electrochemical immunosensor for point-of-care COVID-19 diagnosis using graphene screen-printed electrodes coupled with a 3D-printed portable potentiostat.

A severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a cause of worldwide Coronavirus 2019 (COVID-19) disease pandemic. It is thus important to develop ultra-sensitive, rapid and easy-to-use methods for the identification of COVID-19 infected patients. Herein, an alternative electrochemical immunosensor based on poly(pyrrolepropionic acid) (pPPA) modified graphene screen-printed electrode (GSPE) was proposed for rapid COVID-19 detection. The method was based on a competitive enzyme immunoassay process utilizing horseradish peroxidase (HRP)-conjugated SARS-CoV-2 as a reporter binding molecule to compete binding with antibody against the SARS-CoV-2 receptor binding domain (SARS-CoV-2 RBD) protein. This strategy enhanced the current signal via the enzymatic reaction of HRP-conjugated SARS-CoV-2 RBD antibody on the electrode surface. The modification, immobilization, blocking, and detection processes were optimized and evaluated by amperometry. The quantitative analysis of SARS-CoV-2 was conducted based on competitive enzyme immunoassay with amperometric detection using a 3D-printed portable potentiostat for point-of-care COVID-19 diagnosis. The current measurements at -0.2 V yielded a calibration curve with a linear range of 0.01-1500 ng mL-1 (r2 = 0.983), a low detection limit of 2 pg mL-1 and a low quantification limit of 10 pg mL-1. In addition, the analyzed results of practical samples using the developed method were successfully verified with ELISA and RT-PCR. Therefore, the proposed portable electrochemical immunosensor is highly sensitive, rapid, and reliable. Thus, it is an alternative ready-to-use sensor for COVID-19 point-of-care diagnosis.

Paper-Based Screen-Printed Ionic-Liquid/Graphene Electrode Integrated with Prussian Blue/MXene Nanocomposites Enabled Electrochemical Detection for Glucose Sensing.

As glucose biosensors play an important role in glycemic control, which can prevent the diabetic complications, the development of a glucose sensing platform is still in needed. Herein, the first proposal on the in-house fabricated paper-based screen-printed ionic liquid/graphene electrode (SPIL-GE) modified with MXene (Ti3C2Tx), prussian blue (PB), glucose oxidase (GOx), and Nafion is reported. The concentration of PB/Ti3C2Tx was optimized and the optimal detection potential of PB/Ti3C2Tx/GOx/Nafion/SPIL-GE is -0.05 V. The performance of PB/Ti3C2Tx/GOx/Nafion modified SPIL-GE was characterized by cyclic voltammetry and chronoamperometry technique. This paper-based platform integrated with nanomaterial composites were realized for glucose in the range of 0.0-15.0 mM with the correlation coefficient R2 = 0.9937. The limit of detection method and limit of quantification were 24.5 µM and 81.7 µM, respectively. In the method comparison, this PB/Ti3C2Tx/GOx/Nafion/SPIL-GE exhibits a good correlation with the reference hexokinase method. This novel glucose sensing platform can potentially be used for the good practice to enhance the sensitivity and open the opportunity to develop paper-based electroanalytical devices.

Fast, sensitive and selective simultaneous determination of paraquat and glyphosate herbicides in water samples using a compact electrochemical sensor.

We report a new ready-to-use sensor for simultaneous determination of paraquat (PQ) and glyphosate (GLY) based on a graphite screen-printed electrode modified with a dual-molecularly imprinted polymer coated on a mesoporous silica-platinum core. Amino-mesoporous silica nanoparticles (MSN-NH2) were first synthesized by a simple co-condensation method using tetraethyl orthosilicate and 3-aminopropyltrimethoxysilane. PtNPs were then decorated on the surface of MSN-NH2 by chemical reduction. Finally, the dual-MIP was successfully coated on the MSN-PtNP core. This 3D-surface-imprinting strategy enhances the conductivity and monodispersity of the MSN-PtNPs@d-MIP. Quantitative analysis was performed by differential pulse voltammetry with an oxidation current appearing at -0.95 V for PQ and +0.97 V for GLY. The dual-MIP sensor shows good linear calibration curves in the range of 0.025-500 µM for both analytes with detection limits of 3.1 nM and 4.0 nM for PQ and GLY, respectively. The dual-MIP sensor shows high selectivity and specificity, attributed to the increased affinity of the imprinted cavities formed on the polymer film for the target PQ and GLY molecules. The proposed dual-MIP sensor was successfully applied to detect PQ and GLY concentrations simultaneously in water samples. The ready-to-use dual-MIP sensor is well suited for water-quality control and on-site applications without sophisticated instrumentation.

Ready-to-use paraquat sensor using a graphene-screen printed electrode modified with a molecularly imprinted polymer coating on a platinum core.

We propose the fabrication of a novel ready-to-use electrochemical sensor based on a screen-printed graphene paste electrode (SPGrE) modified with platinum nanoparticles and coated with a molecularly imprinted polymer (PtNPs@MIP) for sensitive and cost-effective detection of paraquat (PQ) herbicide. Successive coating of the PtNPs surface with SiO2 and vinyl end-groups formed the PtNPs@MIP. Next, we terminated the vinyl groups with a molecularly imprinted polymer (MIP) shell. MIP was attached to the PtNPs cores using PQ as the template, methacrylic acid (MAA) as the monomer, ethylene glycol dimethacrylate (EGDMA) as the cross-linker, and 2,2'-azobisisobutyronitrile (AIBN) as the initiator. Coating the SPGrE surface with PtNPs@MIP furnished the PQ sensor. We studied the electrochemical mechanism of PQ on the MIP sensor using cyclic voltammetry (CV) experiments. The PQ oxidation current signal appears at -1.08 V and -0.71 V vs. Ag/AgCl using 0.1 M potassium sulfate solution. Quantitative analysis was performed by anodic stripping voltammetry (ASV) using a deposition potential of -1.4 V for 60 s and linear sweep voltammetric stripping. The MIP sensor provides linearity from 0.05 to 1000 µM (r2 = 0.999), with a lower detection limit of 0.02 µM (at -0.71 V). The compact imprinted sensor gave a highly sensitive and selective signal toward PQ. The ready-to-use MIP sensor can provide an alternative approach to the determination of paraquat residue on vegetables and fruits for food safety applications.

A simple label-free electrochemical sensor for sensitive detection of alpha-fetoprotein based on specific aptamer immobilized platinum nanoparticles/carboxylated-graphene oxide.

A label-free electrochemical aptamer-based sensor has been fabricated for alpha-fetoprotein (AFP) detection. Platinum nanoparticles on carboxylated-graphene oxide (PtNPs/GO-COOH) modified screen-printed graphene-carbon paste electrode (SPGE) was utilized as an immobilization platform, and the AFP aptamer was employed as a bio-recognition element. The synthesized GO-COOH helps to increase the surface area and amounts of the immobilized aptamer. Subsequently, PtNPs are decorated on GO-COOH to enhance electrical conductivity and an oxidation current of the hydroquinone electrochemical probe. The aptamer selectively interacts with AFP, causing a decrease in the peak current of the hydroquinone because the binding biomolecules on the electrode surface hinder the electron transfer of the redox probe. Effects of aptamer concentration and AFP incubation time were studied, and the current changes of the redox probe before and after AFP binding were investigated by square wave voltammetry. The developed aptasensor provides a linear range from 3.0-30 ng mL-1 with a detection limit of 1.22 ng mL-1. Moreover, the aptamer immobilized electrode offers high selectivity to AFP molecules, good stability, and sensitive determination of AFP in human serum samples with high recoveries.

Graphene-based electrochemical genosensor incorporated loop-mediated isothermal amplification for rapid on-site detection of Mycobacterium tuberculosis.

Tuberculosis (TB) is one of the most contagious and lethal infectious diseases that affects more than 10 million individuals worldwide. A lack of rapid TB diagnosis is partly responsible for its alarming spread and prevalence in many regions. To address this problem, we report a novel integrated point-of-care platform to detect a TB-causative bacterium, Mycobacterium tuberculosis (Mtb). This leverages loop-mediated isothermal amplification (LAMP) for Mtb-DNA amplification and the screen-printed graphene electrode (SPGE) for label-free electrochemical analysis of DNA amplicons. When implemented on a portable potentiostat device developed in-house, the system (LAMP-EC) offers a rapid end-point qualitative analysis of specific DNA amplicons that will be displayed as a discrete positive/negative readout on the LCD screen. Under optimized conditions, LAMP-EC showed a comparable detection limit to the previously developed LAMP assay with a lateral flow readout at 1 pg total DNA or 40 Mtb genome equivalents. This highly specific technique detected the presence of TB in all 104 blinded sputum samples with a 100% accuracy. Our technique can also easily be clinically adopted due to its affordability (∼USD2.5/test), rapidity (<65 min turnaround time) and feasibility (lack of advanced instrumental requirement). This serves as a practical incentive, appealing to users in both high- and low-resource settings across the TB endemic regions and economic backgrounds.

Non-Enzymatic Amperometric Glucose Sensor Based on Carbon Nanodots and Copper Oxide Nanocomposites Electrode.

In this research work, a non-enzymatic amperometric sensor for the determination of glucose was designed based on carbon nanodots (C-dots) and copper oxide (CuO) nanocomposites (CuO-C-dots). The CuO-C-dots nanocomposites were modified on the surface of a screen-printed carbon electrode (SPCE) to increase the sensitivity and selectivity of the glucose sensor. The as-synthesized materials were further analyzed for physico-chemical properties through characterization tools such as transmission electron microscopy (TEM) and Fourier-transform infrared spectroscopy (FTIR); and their electrochemical performance was also studied. The SPCE modified with CuO-C-dots possess desirable electrocatalytic properties for glucose oxidation in alkaline solutions. Moreover, the proposed sensing platform exhibited a linear range of 0.5 to 2 and 2 to 5 mM for glucose detection with high sensitivity (110 and 63.3 µA mM-1cm-2), and good selectivity and stability; and could potentially serve as an effective alternative method of glucose detection.

Novel amperometric flow-injection analysis of creatinine using a molecularly-imprinted polymer coated copper oxide nanoparticle-modified carbon-paste-electrode.

We report a novel amperometric flow-injection (FI) analysis of creatinine based on a sensor comprising copper oxide nanoparticles (CuO) coated with a molecularly-imprinted polymer (CuO@MIP) and decorating a carbon-paste electrode (CPE) to form the CuO@MIP/CPE electrode. The CuO@MIP was synthesized by using CuO as the supporting core, creatinine as the template, methacrylic acid (MAA) as monomer, N, N'-(1,2-dihydroxyethylene)bis(acrylamide) (DHEBA) as cross-linker, and 2,2'-azobis (2-methylpropionitrile) (AIBN) as initiator. Morphology and structural characterization reveal that CuO nanoparticle imprinted sites (CuO) synthesized using a precipitation method, exhibits features that are well suited to creatinine detection: high surface area, good analyte diffusion and adsorption characteristics that provide shorter response times, and large numbers of specific cavities for enhanced analyte capacity and sensitivity. Cyclic voltammetric measurements indicate that our sensor provides excellent performance toward electro-oxidation of creatinine. The amperometric FI system was used to quantitatively determine creatinine at the CuO@MIP/CPE sensor, in a phosphate buffer carrier. The imprinted sensor exhibits excellent performance for creatinine oxidation at an applied potential of +0.35 V and flow rate of 0.6 mL.min-1. The as-prepared sensor exhibits a linear dynamic range for creatinine detection from 0.5 to 200 µM (r2 = 0.995) with a limit of detection of 0.083 µM (S/N = 3). The system exhibits satisfactorily good precision (%RSD = 1.94%, n = 30) and selectivity toward creatinine. There is only approximately 20% loss from initial response after 2 weeks when stored at 4 oC. We successfully applied the FI detection system to detect creatinine in human urine samples.

Point-of-care rapid detection of Vibrio parahaemolyticus in seafood using loop-mediated isothermal amplification and graphene-based screen-printed electrochemical sensor.

Vibrio parahaemolyticus is one of the most important foodborne pathogens that cause various life-threatening diseases in human and animals. Here, we present a rapid detection platform for V. parahaemolyticus by combining loop-mediated isothermal amplification (LAMP) and disposable electrochemical sensors based on screen-printed graphene electrodes (SPGEs). The LAMP reactions using primers targeting V. parahaemolyticus toxR gene were optimized at an isothermal temperature of 65 °C, providing specific detection of V. parahaemolyticus within 45 min at the detection limit of 0.3 CFU per 25 g of raw seafood. The LAMP amplicons can be effectively detected using unmodified SPGEs, redox active molecules namely Hoechst-33258 and a portable potentiostat. Therefore, the proposed system is particularly suitable as a point-of-care device for on-site detection of foodborne pathogens.

Electrochemical detection on electrowetting-on-dielectric digital microfluidic chip.

In this work, the use of three-electrode electrochemical sensing system with an electrowetting-on-dielectric (EWOD) digital microfluidic device is reported for quantitative analysis of iodide. T-junction EWOD mixer device was designed using arrays of 50-µm spaced square electrodes for mixing buffer reagent and analyte droplets. For fabrication of EWOD chips, 5-µm thick silver EWOD electrodes were formed on a glass substrate by means of sputtering and lift-off process. PDMS and Teflon thin films were then coated on the electrodes by spin coating to yield hydrophobic surface. An external three-electrode system consisting of Au working, Ag reference and Pt auxiliary wires were installed over EWOD electrodes at the end of T-junction mixer. In experiment, a few-microliter droplets of Tris buffer and iodide solutions were moved toward the mixing junction and transported toward electrochemical electrodes by EWOD process. A short processing time within seconds was achieved at EWOD applied voltage of 300V. The analyte droplets mixed with different concentrations were successfully analyzed by cyclic voltametry. Therefore, the combination of EWOD digital microfluidic and electrochemical sensing system has successfully been demonstrated for rapid chemical analysis with minimal reagent consumption.

AAO-CNTs electrode on microfluidic flow injection system for rapid iodide sensing.

In this work, carbon nanotubes (CNTs) nanoarrays in anodized aluminum oxide (AAO-CNTs) nanopore is integrated on a microfluidic flow injection system for in-channel electrochemical detection of iodide. The device was fabricated from PDMS (polydimethylsiloxane) microchannel bonded on glass substrates that contains three-electrode electrochemical system, including AAO-CNTs as a working electrode, silver as a reference electrode and platinum as an auxiliary electrode. Aluminum, stainless steel catalyst, silver and platinum layers were sputtered on the glass substrate through shadow masks. Aluminum layer was then anodized by two-step anodization process to form nanopore template. CNTs were then grown in AAO template by thermal chemical vapor deposition. The amperometric detection of iodide was performed in 500-µm-wide and 100-µm-deep microchannels on the microfluidic chip. The influences of flow rate, injection volume and detection potential on the current response were optimized. From experimental results, AAO-CNTs electrode on chip offers higher sensitivity and wider dynamic range than CNTs electrode with no AAO template.

Flow injection based microfluidic device with carbon nanotube electrode for rapid salbutamol detection.

A microfabicated flow injection device has been developed for in-channel electrochemical detection (ECD) of a beta-agonist, namely salbutamol. The microfluidic system consists of PDMS (polydimethylsiloxane) microchannel and electrochemical electrodes formed on glass substrate. The carbon nanotube (CNT) on gold layer as working electrode, silver as reference electrode and platinum as auxiliary electrode were deposited on a glass substrate. Silver, platinum, gold and stainless steel catalyst layers were coated by DC-sputtering. CNTs were then grown on the glass substance by thermal chemical vapor deposition (CVD) with gravity effect and water-assisted etching. 100-microm-deep and 500-microm-wide PDMS microchannels fabricated by SU-8 molding and casting were then bonded on glass substrate by oxygen plasma treatment. Flow injection and ECD of salbutamol was performed with the amperometric detection mode for in-channel detection of salbutamol. The influences of flow rate, injection volume, and detection potential on the response of current signal were optimized. Analytical characteristics, such as sensitivity, repeatability and dynamic range have been evaluated. Fast and highly sensitive detection of salbutamol have been achieved. Thus, the proposed combination of the efficient CNT electrode and miniaturized lab-on-a-chip is a powerful platform for beta-agonists detection.

High sensitivity electrochemical cholesterol sensor utilizing a vertically aligned carbon nanotube electrode with electropolymerized enzyme immobilization.

In this report, a new cholesterol sensor is developed based on a vertically aligned CNT electrode with two-step electrochemical polymerized enzyme immobilization. Vertically aligned CNTs are selectively grown on a 1 mm(2) window of gold coated SiO(2)/Si substrate by thermal chemical vapor deposition (CVD) with gravity effect and water-assisted etching. CNTs are then simultaneously functionalized and enzyme immobilized by electrochemical polymerization of polyaniline and cholesterol enzymes. Subsequently, ineffective enzymes are removed and new enzymes are electrochemically recharged. Scanning electron microscopic characterization indicates polymer-enzyme nanoparticle coating on CNT surface. Cyclic voltammogram (CV) measurements in cholesterol solution show the oxidation and reduction peaks centered around 450 and -220 mV, respectively. An approximately linear relationship between the cholesterol concentration and the response current could be observed in the concentration range of 50-300 mg/dl with a sensitivity of approximately 0.22 µA/mg·dl(-1), which is considerably higher compared to previously reported CNT bioprobe. In addition, good specificity toward glucose, uric acid acetaminophen and ascorbic acid have been obtained. Moreover, sensors have satisfactory stability, repeatability and life time. Therefore, the electropolymerized CNT bioprobe is promising for cholesterol detection in normal cholesterol concentration in human blood.