Utilizing COVID-19 as a Model for Diagnostics Using an Electrochemical Sensor.

This paper reports a rapid and sensitive sensor for the detection and quantification of the COVID-19 N-protein (N-PROT) via an electrochemical mechanism. Single-frequency electrochemical impedance spectroscopy was used as a transduction method for real-time measurement of the N-PROT in an immunosensor system based on gold-conjugate-modified carbon screen-printed electrodes (Cov-Ag-SPE). The system presents high selectivity attained through an optimal stimulation signal composed of a 0.0 V DC potential and 10 mV RMS-1 AC signal at 100 Hz over 300 s. The Cov-Ag-SPE showed a log response toward N-PROT detection at concentrations from 1.0 ng mL-1 to 10.0 µg mL-1, with a 0.977 correlation coefficient for the phase (θ) variation. An ML-based approach could be created using some aspects observed from the positive and negative samples; hence, it was possible to classify 252 samples, reaching 83.0, 96.2 and 91.3% sensitivity, specificity, and accuracy, respectively, with confidence intervals (CI) ranging from 73.0 to 100.0%. Because impedance spectroscopy measurements can be performed with low-cost portable instruments, the immunosensor proposed here can be applied in point-of-care diagnostics for mass testing, even in places with limited resources, as an alternative to the common diagnostics methods.

An electrochemical microfluidic device for non-enzymatic cholesterol determination using a lab-made disposable electrode.

Cholesterol is an important steroid and hormone precursor, and its levels in the blood are associated with risk factors for cardiovascular diseases. In this work, a non-enzymatic methodology for cholesterol determination in serum samples is described. First, a working electrode was constructed using homemade ink and a plastic substrate by a simple dunking process. Next, the dunked electrode (DWE) was modified with nickel ions (Ni-DWE) and combined with a low-cost microfluidic platform, resulting in a thread-based electroanalytical device (µTED). The arrangement of µTED consists of two coupled electrodes (one reference in the inlet reservoir and an auxiliary electrode against the outlet reservoir) and a mobile support for facile working electrode exchange. After optimization of construction parameters, the system was applied for non-enzymatic determination of cholesterol under alkaline conditions using the redox pair Ni(II)/Ni(III) as a mediator. Under the best analytical conditions, a calibration curve was constructed with a linear dynamic range (LDR) from 0.25 to 25.0 µmol L-1, and the calculated limits of detection (LOD) and quantification (LOQ) were 0.074 and 0.24 µmol L-1, respectively. No effects of possible interferents on electrochemical response were found in the presence of ascorbic acid, uric acid, dopamine, cysteine, and glucose, suggesting that the proposed device can be used for the determination of cholesterol without significant matrix effects of human plasma. Finally, cholesterol analysis was carried out using spiked plasma samples, and good recovery values were achieved.

Enhancing a SARS-CoV-2 nucleocapsid antigen test sensitivity with cost efficient strategy through a cotton intermembrane insertion.

Lateral flow antigen tests have been widely used in the Covid-19 pandemic, allowing faster diagnostic test results and preventing further viral spread through isolation of infected individuals. Accomplishment of this screening must be performed with tests that show satisfactory sensitivity in order to successfully detect the target protein and avoid false negatives. The aim of this study was to create a lateral flow test that could detect SARS-CoV-2 nucleocapsid protein in low concentrations that were comparable to the limits of detection claimed by existing tests from the market. To do so, several adjustments were necessary during research and development of the prototypes until they were consistent with these criteria. The proposed alternatives of increasing the test line antibody concentration and addition of an intermembrane between the conjugate pad and the nitrocellulose membrane were able to increase the sensitivity four-fold and generate a new rapid test prototype called "lateral flow intermembrane immunoassay test" (LFIIT). This prototype showed an adequate limit of detection (2.0 ng mL-1) while maintaining affordability and simplicity in manufacturing processes.

Biochar: An environmentally friendly platform for construction of a SARS-CoV-2 electrochemical immunosensor.

Waste management is a key feature to ensure sustainable consumption and production patterns, and to combat the impacts of climate change. In this scenario, the production of biochar from different biomasses results in environmental and economic advantages. In this study, biochar was produced from sugarcane bagasse pyrolysis, to immobilize biomolecules, in order to assemble an electrochemical immunosensor to detect antibodies against SARS-CoV-2. For this, screen-printed carbon electrodes (SPCE) were modified with a dispersion of biochar and used to immobilize the receptor-binding-domain (RBD) against virus S-protein, through EDC/NHS crosslinking reaction. Under the best set of experimental conditions, negative and positive serum samples responses distinguished based on a cutoff value of 82.3 %, at a 95 % confidence level. The immunosensor showed selective behavior to antibodies against yellow fever and its performance was stable up to 7 days of storage. Therefore, biochar yielded from sugarcane bagasse is an ecofriendly material that can be used as a platform to immobilize biomolecules for construction of electrochemical biosensors.

Selective carbonaceous-based (nano)composite sensors for electrochemical determination of paraquat in food samples.

A novel electrochemical sensor based on activated biochar (AB4) and reduced graphene oxide (rGO) was developed and tested for detection of paraquat (PQ) in food samples. Precursor biochar was obtained by the pyrolysis of water hyacinth biomass at 400, 500, and 600 °C, followed by a chemical activation step using HNO3 to increase the amount of oxygenated and nitrogenated groups. The modified electrodes (rGO-AB4) were tested in different experimental conditions, and exhibited good response under the optimized conditions, showing linearity from 0.74 to 9.82 µmol L-1 and a limit of detection and limit of quantification of 0.02 µmolL-1 and 0.07 µmol L-1, respectively. Interfering species such as glyphosate caused insignificant changes in the peak current of paraquat, and the selectivity of the method was tested using blank and spiked samples of coconut water, wastewater, honey, lettuce and lemon. Recovery ranged from 87.70±2.07% to 103.80±3.94%.

Label-free aptasensor for p24-HIV protein detection based on graphene quantum dots as an electrochemical signal amplifier.

Human immunodeficiency virus (HIV) is still considered a pandemic, and the detection of p24-HIV protein has an important role in the early diagnosis of HIV in adults and newborns. The accessibility of these trials depends on the price and execution difficulty of the method, which can be reduced using electrochemical methods by using enzymeless approaches, disposable and accurate devices. In this work, graphene quantum dots were acquired by a simple synthesis and employed as an electrochemical signal amplifier and support for the aptamer immobilization through a feasible and stable modification of disposable screen-printed electrodes. The device has been easily assembled and used to detect p24-HIV protein without the interference of similar proteins or sample matrix. Using the best set of experimental conditions, a linear correlation between analytical signal and log of p24-HIV concentration from 0.93 ng mL-1 to 93 µg mL-1 and a limit of detection of 51.7 pg mL-1 were observed. The developed device was applied to p24 determination in spiked human serum and provided distinct levels of signal for positive and negative samples, successfully identifying real samples with the target protein. This sensor is a step towards the development of point-of-care devices and the popularization of electrochemical methods for trials and diagnostics of relevant diseases.

Electrochemical sensor based on biochar and reduced graphene oxide nanocomposite for carbendazim determination.

For the first time, a nanocomposite based on biochar and reduced graphene oxide (rGO) was employed to construct a modified carbon paste electrode and applied for the determination of carbendazim (CBZ). Biochar was obtained by through pyrolysis of Eichhornia crassipes biomass, also known how "Aguapé" at 400 °C. The modified electrode with our nanocomposite proposal shows to be able to preconcentrate CBZ and presented the highest analytical response in comparison to the unmodified electrode and by the electrodes prepared with the proposed materials separately. Using differential pulse voltammetry (DPV) under optimized conditions, the sensor showed a linear dynamic response (LDR) from 30 to 900 nmol L-1, a limit of detection (LOD) of 2.3 nmol L-1 and limit of quantification (LOQ) of 7.7 nmol L-1. No significant influence of inorganic ions or organic compounds on sensor response was verified, considering the recovery evaluation data. The proposed sensor was successfully applied for the determination of CBZ in spiked whole orange juice, lettuce leaves, drinking water, and wastewater samples. Good recovery values were found using the ex-situ methodology, showing excellent analytical performance of the electrochemical sensor based on biochar and rGO nanocomposite.

A simple enzymeless approach for Paraoxon determination using imidazole-functionalized carbon nanotubes.

This work describes the application of a glassy carbon electrode (GCE) modified with imidazole functionalized carbon nanotubes (CNT-H-IMZ) for Paraoxon (PX) determination in samples of commercial, fresh and 100% orange juice. Homemade multi-walled CNTs were treated according to the Hummers procedure to oxidize graphite and later chemically functionalized with imidazole groups. Modified electrodes with CNT-H-IMZ presented a high peak current of PX reduction and an electrocatalytic effect in comparison to the other electrodes. This behavior was associated with the synergistic contribution of IMZ and CNT that increases the electrochemical activity of PX. Repeatability and reproducibility studies showed that the relative peak current values did not show significant differences between them, less than 10%, and it was possible to define that the diffusional process is the mechanism that limits the electrode mass transport. After the optimization of parameters inherent to the methodology and the voltammetric technique, the proposed device presented a linear region of 1.0 to 16.0 µM-1 (R2 = 0.99), presenting LOD and LOQ as 120 and 400 nM-1, respectively. The method proposed was successfully applied to PX determination in spiked samples.

A carbon fiber ultramicroelectrode as a simple tool to direct antioxidant estimation based on caffeic acid oxidation.

This work describes the construction and evaluation of carbon fiber ultramicroelectrodes (CF-UMEs) in the voltammetric estimation of the antioxidant capacity of wine and grape samples based on caffeic acid (HCAF) oxidation. For this, lab-made CF-UMEs were constructed using an arrangement of six carbon fibers (7 µm diameters individual) assembled in a glass capillary, and caffeic acid (HCAF) was used as a standard solution. By using the most straightforward 2-electrode cell arrangement (the CF-UME as a working electrode and Ag/AgCl as a reference/auxiliary electrode), voltammetric measurements of a 1.0 mmol L-1 HCAF solution were done in the absence of a supporting electrolyte. A sigmoidal voltammetric profile was observed in CF-UMEs caused by a more effective mass transport by radial diffusion, which leads to a rapid formation of the diffusion layer. Reproducibility studies for different 6-fiber electrodes manually constructed in different batches showed an RSD of less than 5%. For the same electrode surface, a variation of 2.7% was observed. Under optimized conditions, a linear relationship between anodic peak current and HCAF concentration from 3.0 to 500 µmol L-1 with a sensitivity of 12 µA L mol-1 was reached. The limits of detection (LOD) and quantification (LOQ) were calculated to be 0.41 and 1.26 µmol L-1, respectively. The proposed electrochemical method was applied in the estimation of the antioxidant capacity in three different wine samples as well as in green and red grapes. Concordant and satisfactory results by comparison with a proper method were obtained, which suggests that the proposed sensor can be successfully applied for direct analysis of wine and grape samples by estimation of HCAF content.

Nonenzymatic electrochemical sensor based on imidazole-functionalized graphene oxide for progesterone detection.

The modification of electrode surfaces has been the target of study for many researchers in order to improve the analytical performance of electrochemical sensors. Herein, the use of an imidazole-functionalized graphene oxide (GO-IMZ) as an artificial enzymatic active site for voltammetric determination of progesterone (P4) is described for the first time. The morphology and electrochemical performance of electrode modified with GO-IMZ were characterized by scanning electron microscopy and cyclic voltammetry, respectively. Under optimized conditions, the proposed sensor showed a synergistic effect of the GO sheets and the imidazole groups anchored on its backbone, which promoted a significant enhancement on electrochemical reduction of P4. Figures of merits such as linear dynamic response for P4 concentration ranging from 0.22 to 14.0 µmol L-1, limit of detection of 68 nmol L-1 and limit of quantification and 210 nmol L-1 were found. In addition, presented a higher sensitivity, 426 nA L µmol-1, when compared to the unmodified electrode. Overall, the proposed device showed to be a promising platform for a simple, rapid, and direct analysis of progesterone.

Evaluation of antimony microparticles supported on biochar for application in the voltammetric determination of paraquat.

This work describes the construction and application of carbon paste electrodes modified with biochar and antimony microparticles (SbBCPE) for voltammetric determination of paraquat using a simple and sensitive procedure based on voltammetric stripping analysis. Some parameters such as amount of biochar and antimony used in the composition of the carbon paste and instrumental parameters were examined in detail. Under optimized conditions, an analytical curve was obtained for paraquat determination employing SbBCPE, which showed a linear response ranging from 0.2 to 2.9 µmol L(-1), with limit of detection and quantification of 34 nmol L(-1) and 113 nmol L(-1), respectively, after paraquat pre-concentration of 120 s. The repeatability study presented a RSD=2.0% for 10 consecutive measurements using the same electrode surface and the reproducibility study showed a RSD=2.7% for measurements with 10 different electrode surfaces. The proposed sensor was successfully applied for paraquat determination in tap water and citric fruit juice spiked samples and good recoveries were obtained without any sample pre-treatment, showing its promising analytical performance.