An Electrochemical o-Phthalaldehyde Sensor Using a Modified Disposable Screen-Printed Electrode with Polyacrylate Hydrogel for Concentration Verification of Clinical Disinfectant.

The study proposes an o-phthalaldehyde (OPA) sensor for rapid and reliable detection of OPA in healthcare disinfection practices, based on a hydrogel-modified screen-printed carbon electrode strip. The hydrogel film, which contains glycine and N-acetylcysteine, reacts with OPA to produce a reductive isoindole derivative. The derivative is then oxidized for OPA determination using cyclic voltammetry. The proposed sensor achieves an optimal detection time of 20-30 s and requires only a small analyte volume of 5 µL. It exhibits good precision (10%) and sensitivity (3.3 µA/cm2 mM) in a phosphate-buffered solution (pH 7.6), with excellent linearity (R2 > 0.97) and precision (<3%) in the detection range (0.2-0.6%) required for clinical OPA solutions. Moreover, the sensor demonstrates good concentration verification of Cidex-OPA disinfection in healthcare institutes, with high sensitivity (18.28 µA/cm2 mM) and precision around the minimum effective concentration (0.3%). Overall, the proposed sensor offers a promising and practical solution for accurate and reliable OPA detection in clinical disinfection practices.

Ultrasensitive Determination of Glial-Fibrillary-Acidic-Protein (GFAP) in Human Serum-Matrix with a Label-Free Impedimetric Immunosensor.

In this work, immobilizing anti-GFAP antibodies via covalent attachment onto L-cysteine/gold nanoparticles that were modified with screen-printed carbon electrodes (Anti-GFAP/L-cys/AuNps/SPCE) resulted in the development of a sensitive label-free impedance immunosensor for the detection of Glial Fibrillary Acidic Protein (GFAP). The immunosensor's stepwise construction was studied using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). L-cysteine was chosen as the linker between GFAP antibodies and Au NPs/SPCE because it enables the guided and stable immobilization of GFAP antibodies, thus resulting in increased immunosensor sensitivity. As a redox probe, 5 mM of [Fe(CN)6]3-/4- was used to measure the electron-transfer resistance (Ret), which was raised by the binding of antigens to the immobilized anti-GFAP on the surface of the modified electrode. A linear correlation between Rct and GFAP concentration was achieved under optimum conditions in the range of 1.0-1000.0 pg/mL, with an extraordinarily low detection limit of 51.0 fg/mL. The suggested immunosensor was successfully used to detect the presence of GFAP in human blood serum samples, yielding good findings. As a result, the proposed platform may be utilized to monitor central nervous system injuries.

Impedance Technique-Based Label-Free Electrochemical Aptasensor for Thrombin Using Single-Walled Carbon Nanotubes-Casted Screen-Printed Carbon Electrode.

An impedance technique-based aptasensor for the detection of thrombin was developed using a single-walled carbon nanotube (SWCNT)-modified screen-printed carbon electrode (SPCE). In this work, a thrombin-binding aptamer (TBA) as probe was used for the determination of thrombin, and that was immobilized on SWCNT through π-π interaction. In the presence of thrombin, the TBA on SWCNT binds with target thrombin, and the amount of TBA on the SWCNT surface decreases. The detachment of TBA from SWCNT will be affected by the concentration of thrombin and the remaining TBA on the SWCNT surface can be monitored by electrochemical methods. The TBA-modified SWCNT/SPCE sensing layer was characterized by cyclic voltammetry (CV). For the measurement of thrombin, the change in charge-transfer resistance (Rct) of the sensing interface was investigated using electrochemical impedance spectroscopy (EIS) with a target thrombin and [Fe(CN)6]3- as redox maker. Upon incubation with thrombin, a decrease of Rct change was observed due to the decrease in the repulsive interaction between the redox marker and the electrode surface without any label. A plot of Rct changes vs. the logarithm of thrombin concentration provides the linear detection ranges from 0.1 nM to 1 µM, with a ~0.02 nM detection limit.

Glucose oxidase immobilized amine terminated multiwall carbon nanotubes/reduced graphene oxide/polyaniline/gold nanoparticles modified screen-printed carbon electrode for highly sensitive amperometric glucose detection.

A novel amine terminated multiwall carbon nanotubes/polyaniline/reduced graphene oxide/gold nanoparticles modified screen-printed carbon electrode (SPCE) was fabricated. Followed by, glucose oxidase (GOx) was immobilized on SPCE for highly sensitive glucose biosensor. The synthesized nanomaterial and their composites were characterized using scanning electron microscope (SEM) and UV-Visible spectroscopy. The electrochemical analysis has been followed at different stages of glucose oxidase coating on modified SPCE using cyclic voltammetry. The reduction current has enhanced 13.43 times with the lowest working potential by the modified SPCE when compared to bare SPCE. The glucose biosensor exhibited good reproducibility (90.23%, n = 7), high stability (after 30 days 96% at -20 °C storage, 2 week 74.5% at -4 °C storage), wide linear range (1-10 mM), less KMapp value (0.734), lowest detection limit (64 µM) and good sensitivity (246 µ Acm-2 mM-1). The biosensor was validated for the detection of glucose level in human blood serum samples using the amperometric technique. As designed nanocomposite based SPCE has the potential for an efficient glucose sensor, which also enabled the platform for various biochemical sensors.

Voltammetric determination of iodide in iodized table salt using cetyltrimethylammonium bromide as ion-pairing.

In this work, voltammetric study based on cetyltrimethylammonium bromide (CTAB) as an ion-pairing agent for the determination of iodine level in iodized table salt has been explored. CTAB was used as an intermediate compound between iodide (I-) and the electrode due to its ability to dissociate to produce cetyltrimethylammonium ions ([CTA]+). The [CTA]+ with a long hydrophobic alkyl chain can be directly adsorbed onto the surface of the working electrode, and this in turns coated the electrode with cationic charge and enhance the electrode ability to bind to iodide (I-) and other molecular iodine ions. A mixture of iodide and CTAB ([CTA]+I-) was prepared and potential of 1.0 V for 60.0 s was applied to pre-concentrate the solution on the working electrode causing the [CTA]+I- to oxidize to iodine (I2). The produced I2 immediately react with chloride ion (Cl-) from the electrolyte of hydrochloric acid (HCl) to produce I2Cl- and form ion-pair with CTA+ as [CTA]+I2Cl-. The linear calibration curve of the developed method towards iodide was in the concentration range of 0.5-4.0 mg/L with sensitivity of - 1.383 µA mg/L-1 cm-2 (R2 = 0.9950), limit of detection (LOD) of 0.3 mg/L and limit of quantification (LOQ) of 1.0 mg/L, respectively. The proposed method indicates good agreement with the standard method for iodine determination with recovery range from 95.0 to 104.3%. The developed method provided potential application as a portable on-site iodine detector.

Development of a disposable electrochemical sensor for detection of cholesterol using differential pulse voltammetry.

In this study, a sensitive and selective electrochemical sensor was fabricated by using a screen printed carbon electrode (SPCE), multi-walled carbon nanotubes (MWCNTs) and ß-cyclodextrin (ß-CD) for detecting cholesterol. MWCNTs were functionalized with benzoic acid moiety by employing diazonium salt chemistry, and, subsequently, a thin film of functionalized CNTs were coated on the surface of SPCE. Afterwards, ß-CD was immobilized on functionalized MWCNTs modified SPCE which acts as a host to recognize guest (cholesterol) molecule specifically. Under the optimal experimental conditions and using differential pulse voltammetry (DPV) as transduction technique the sensor was able to detect cholesterol level ranges from 1 nM to 3 µM, with a detection limit of 0.5 nM. Specificity of the developed sensor towards target analyte (cholesterol) was confirmed in the presence of common interfering species including glucose, uric acid and ascorbic acid. The applicability of proposed sensor was also demonstrated for cholesterol determination in human serum samples with good recovery results (94-96%) and maximum RSD (relative standard deviation) of 4.5%.

Ultrasensitive determination of receptor tyrosine kinase with a label-free electrochemical immunosensor using graphene quantum dots-modified screen-printed electrodes.

A new label-free electrochemical immunosensor is constructed for the selective and sensitive determination of the clinically relevant biomarker receptor tyrosine kinase (AXL) in human serum. The disposable immunosensing platform is prepared by immobilization of the specific anti-AXL antibody onto amine functionalized graphene quantum dots (fGQDs)-modified screen-printed carbon electrodes (SPCEs). The affinity reactions were monitored by measuring the decrease in the differential pulse voltammetric (DPV) response of the redox probe Fe(CN)63-/4-. All the experimental variables involved in the preparation of the modified electrodes and in the immunosensor performance were optimized. The as prepared immunosensor exhibits an improved analytical performance with respect to other electrochemical immunosensors reported so far, with a wider range of linearity and a lower detection limit, 0.5 pg mL-1, which is more than one hundred thousand times lower than the established cut-off value for heart failure (HF) diagnosis in serum (71 ng mL-1). The developed immunosensor was successfully applied to the determination of the endogenous content of AXL in serum of HF patients without any matrix effect observed after just a sample dilution.

Sequential injection differential pulse voltammetric method based on screen printed carbon electrode modified with carbon nanotube/Nafion for sensitive determination of paraquat.

The screen-printed carbon electrode (SPCE) modified with various nanoparticles has been studied for using as a working electrode in voltammetric technique. The electrochemical behavior of paraquat on different electrodes was studied by cyclic voltammetry (CV), and then differential pulse voltammetry (DPV) has been employed for trace analysis of paraquat based on redox reaction which the peak current was directly proportional to the concentration of paraquat in the solution. The SPCE modified with carbon nanotube dispersed in Nafion and ethanol (SPCE-CNT/Nafion) gave the best result. Sequential injection-differential pulse voltammetric (SI-DPV) method has been developed for more automated analysis and to reduce chemical consumption. The parameters affecting the SI-DPV system such as step potential, modulation amplitude, flow rate, and concentration of sodium chloride as an electrolyte were studied to improve the sensitivity. Under the optimum condition of the system, i.e., Nafion concentration of 1% (w/v), volume of CNT suspension of 2µL, flow rate of 100µLs-1, step potential of 5mV, modulation amplitude of 100mV and concentration of sodium chloride of 1M, a linear calibration graph in the range of 0.54-4.30µM with a good R2 of 0.9955 and a limit of detection of 0.17µM (0.03mgL-1) were achieved. The proposed system shows high tolerance to some possible interfering ions in natural water, surfactant, and other pesticides. The relative standard deviation (RSD) was 4.2% for 11 replicate measurements with the same electrode. The reproducibility for the preparation of 7 modified electrodes was 2.3% RSD. Recoveries of the analysis were obtained in the range of 82-106%. The developed system can be conveniently applied for analysis without pretreatment of the samples.

Electrochemically reduced graphene oxide-modified screen-printed carbon electrodes for a simple and highly sensitive electrochemical detection of synthetic colorants in beverages.

A simple and highly sensitive electrochemical sensor based on an electrochemically reduced graphene oxide-modified screen-printed carbon electrode (ERGO-SPCE) for the simultaneous determination of sunset yellow (SY) and tartrazine (TZ) was proposed. An ERGO film was coated onto the electrode surface using a cyclic voltammetric method and then characterized by scanning electron microscopy (SEM). In 0.1M phosphate buffer at a pH of 6, the two oxidation peaks of SY and TZ appeared separately at 0.41 and 0.70V, respectively. Surprisingly, the electrochemical response remarkably increased approximately 90- and 20-fold for SY and TZ, respectively, using the modified electrode in comparison to the unmodified electrode. The calibration curves exhibited linear ranges from 0.01 to 20.0µM for SY and from 0.02 to 20.0µM for TZ. The limits of detection were found to be 0.50 and 4.50nM (at S/N=3) for SY and TZ, respectively. Furthermore, this detection platform provided very high selectivity for the measurement of both colorants. This electrochemical sensor was successfully applied to determine the amount of SY and TZ in commercial beverages. Comparison of the results obtained from this proposed method to those obtained by an in-house standard technique proved that this developed method has good agreement in terms of accuracy for practical applications. This sensor offers an inexpensive, rapid and sensitive determination. The proposed system is therefore suitable for routine analysis and should be an alternative method for the analysis of food colorants.

Indirect electrochemical detection for total bile acids in human serum.

Bile acids level in serum is a useful index for screening and diagnosis of hepatobiliary diseases. As bile acids concentration is closely related to the degree of hepatobiliary diseases, detecting it is a vital factor to understand the stage of the diseases. The prevalent determination for bile acids is the enzymatic cycling method which has low sensitivity while reagent-consuming. It is desirable to develop a new method with lower cost and higher sensitivity. An indirect electrochemical detection (IED) for bile acids in human serum was established using the screen printed carbon electrode (SPCE). Since bile acids do not show electrochemical signals, they were converted to 3-ketosteroids by 3-α-hydroxysteroid dehydrogenase (3α-HSD) in the presence of nicotinamide adenine dinucleotide (NAD(+)), which was reduced to NADH. NADH could then be oxidized on the surface of SPCE, generating a signal that was used to calculate the total bile acids (TBA) concentration. A good linear calibration for TBA was obtained at the concentration range from 5.00µM to 400µM in human serum. Both the precisions and recoveries were sufficient to be used in a clinical setting. The TBA concentrations in 35 human serum samples by our IED method didn't show significant difference with the result by enzymatic cycling method, using the paired t-test. Moreover, our IED method is reagent-saving, sensitive and cost-effective.

Optimized design of a nanostructured SPCE-based multipurpose biosensing platform formed by ferrocene-tethered electrochemically-deposited cauliflower-shaped gold nanoparticles.

The demand for on-site nanodevices is constantly increasing. The technology development for the design of such devices is highly regarded. In this work, we report the design of a disposable platform that is structured with cauliflower-shaped gold nanoparticles (cfAuNPs) and we show its applications in immunosensing and enzyme-based detection. The electrochemical reduction of Au(III) allows for the electrodeposition of highly dispersed cauliflower-shaped gold nanoparticles on the surface of screen-printed carbon electrodes (SPCEs). The nanostructures were functionalized using ferrocenylmethyl lipoic acid ester which allowed for the tethering of the ferrocene group to gold, which serves as an electrochemical transducer/mediator. The bioconjugation of the surface with anti-human IgG antibody (α-hIgG) or horseradish peroxidase (HRP) enzyme yields biosensors, which have been applied for the selective electrochemical detection of human IgG (hIgG) or H2O2 as model analytes, respectively. Parameters such as the number of sweeps, amount of charge generated from the oxidation of the electrodeposited gold, time of incubation and concentration of the ferrocene derivatives have been studied using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and scanning electron microscopy (SEM). Selectivity and specificity tests have been also performed in the presence of potentially interfering substances to either hIgG or H2O2. Results showed that the devised immunosensor is endowed with good selectivity and specificity in the presence of several folds of competitive analytes. The enzyme-based platform showed a good catalytic activity towards H2O2 oxidation which predestined it to potential applications pertaining to enzymatic kinetics studies. The levels of hIgG in human serum and H2O2 in honey were successfully determined and served as assessment tools of the applicability of the platforms for real samples analysis.

Gold nanoparticles conjugates-amplified aptamer immunosensing screen-printed carbon electrode strips for thrombin detection.

Thrombin plays the role in cardiovascular diseases and regulates many processes in inflammation and could be a feature of many pathological conditions, including the thromboembolic disease, cancer and neurodegenerative diseases. An ultrasensitive and amplified electrochemical sandwich assay using screen-printed carbon electrode (SPCE) strips for thrombin detection was established in this study. The conductivity and sensing performance of the carbon electrodes were enhanced by using gold nanoparticles (AuNPs). The aptamer addressed on the strips was used as a primary probe to capture thrombin in the detected samples. An amplifier was invented for recognizing thrombin captured on the SPCE, which is the multiple molecules of anti-thrombin antibody (Ab) and horseradish peroxidase (HRP) co-modified AuNPs (AuNPs/Ab-HRP). Hydrogen peroxide was used as the substrate for HRP and then the response current (RC) could be detected. The optimization of these AuNPs conjugates-amplified aptamer immunosensing SPCE strips was conducted for thrombin detection. The detection sensitivity showed a linear relation between RC and thrombin concentration in the range of 10 pM-100 nM, and limit of detection (LOD) was 1.5 pM. The fabricated AuNPs/Ab-HRP-amplified aptamer immunosensing SPCE strips were further used to detect thrombin in human serum with a linear range of 100 pM-100 nM. This study provided the promising SPCE strips with highly sensitive and rapid detection for thrombin by the electrochemical aptasensor combined with AuNPs conjugates for amplifying the detection signal.