Development of an electrochemical biosensor utilizing a combined aptamer and MIP strategy for the detection of the food allergen lysozyme.

This study aims to develop a MIP-Apt-based electrochemical biosensor for the sensitive and selective determination of Lysozyme (Lyz), a food allergen. For the development of the sensor, in the first stage, modifications were made to the screen-printed electrode (SPE) surface with graphene oxide (GO) and gold nanoparticles (AuNPs) to increase conductivity and surface area. The advantages of using aptamer (Apt) and molecularly imprinted polymer (MIP) technology were combined in a single biointerface in the prepared sensing tool. Surface characterization of the biosensor was performed using scanning electron microscopy (SEM), atomic force microscopy (AFM), X-ray photoelectron spectrometry (XPS), contact angle measurements, cyclic voltammetry (CV), differential pulse voltammetry (DPV), and electrochemical impedance spectroscopy (EIS). A wide linear range from 0.001 to 100 pM was obtained under optimized conditions for the determination of Lyz detection using the proposed MIP-Apt sensing strategy. The limit of detection (LOD) and limit of quantification (LOQ) for Lyz were 3.67 fM and 12 fM, respectively. This biosensor displays high selectivity, repeatability, reproducibility, and long storage stability towards Lyz detection. The results show that a sensitive and selective sensor fabrication is achieved compared with existing methods.

Analytical studies on some pesticides with antifungal effects: Simultaneous determination by HPLC, investigation of interactions with DNA and DNA damages.

A simple, and fast method was developed for the simultaneous determination of five fungicides, namely thiram (THR), epoxiconazole (EPO), hexaconazole (HEX), tebuconazole (TEB), and diethofencarb (DIE), in different matrices by HPLC-UV. Parameters influencing the peak shape and resolution, such as the composition of mobile phase, pH and concentration of buffer solution, and column temperature, were examined and optimized. The proposed method was validated in terms of linearity, sensitivity, precision, and accuracy. Forced degradation studies were carried out for all analytes to demonstrate the specificity of the method and to evaluate the stability of analytes under different conditions. DNA interaction and DNA damage studies were conducted by HPLC and comet assay, respectively. All fungicides were found to bind DNA, except for DIE. While the binding coefficients for EPO, HEX, and TEB were of the order of 104, THR was found to interact more strongly with DNA with a binding coefficient of higher than 106. DIE did not induce DNA damage at any concentration tested. On the other hand, TEB, HEX, and EPO induced DNA damage up to 30 µg/mL. THR showed cytotoxic effects at 20 and 30 µg/mL and caused significant DNA damage at lower concentrations.

Comprehensive views toward the biomolecular recognition of an anticancer drug, leflunomide with human serum albumin.

Biomolecular association of an anticancer drug, leflunomide (LEF) with human serum albumin (HSA), the leading ligands carrier in human circulation was characterized using biophysical (i.e., fluorescence, absorption and voltammetric) methods and computational (i.e., molecular docking and molecular dynamics simulation) techniques. Evaluations of fluorescence, absorption and voltammetric findings endorsed the complex formation between LEF and HSA. An inverse relationship of Stern-Volmer constant-temperature and hyperchromic shift of the protein's absorption signal with addition of LEF confirmed the LEF quenched the HSA fluorescence through static process. Moderate nature of binding strength (binding constant = 2.76-4.77 × 104 M-1) was detected towards the LEF-HSA complexation, while the association process was naturally driven via hydrophobic interactions, van der Waals interactions and hydrogen bonds, as evident from changes in entropy (ΔS= + 19.91 J mol-1 K-1) and enthalpy (ΔH = - 20.09 kJ mol-1), and molecular docking assessments. Spectral analyses of synchronous and three-dimensional fluorescence validated microenvironmental fluctuations near Trp and Tyr residues upon LEF binding to the protein. LEF association with HSA significantly defended temperature-induced destabilization of the protein. Although LEF was found to attach to HSA at Sudlow's sites I and II, but exhibited greater preference toward its site I, as detected by the investigations of competitive site-marker displacement. Molecular dynamics simulation assessment revealed that the complex attained equilibrium throughout simulations, showing the LEF-HSA complex constancy.Communicated by Ramaswamy H. Sarma.

Evaluation of aptamer and molecularly imprinted polymers as a first hybrid sensor for leptin detection at femtogram levels.

Selective and sensitive determination of macromolecules maintains its importance in diagnosing and determining diseases to protect human health. In this study, a hybrid sensor designed with dual recognition elements consisting of both aptamers (Apt) and molecularly imprinted polymers (MIPs) was carried out for the ultra-sensitive determination of Leptin. Firstly, the screen-printed electrode (SPE) surface was coated with platinum nanospheres (Pt NSs) and gold nanoparticles (Au NPs) to provide immobilization of the Apt[Leptin] complex on the surface. In the next step, the formed polymer layer around the complex using the electropolymerization of orthophenilendiamine (oPD) kept the Apt molecules on the surface more effectively. As expected, a synergistic effect occurred between the formed MIP cavities by removing Leptin from the surface and the embedded Apt molecules to fabricate a hybrid sensor. Under optimal conditions, responses in differential pulse voltammetry (DPV) currents showed a linear response over a wide concentration range from 1.0 fg/mL to 10.0 pg/mL with a limit of detection (LOD) of 0.31 fg/mL for Leptin detection. Moreover, the effectiveness of the hybrid sensor was assessed using real samples, such as human serum and plasma samples, and satisfactory recovery findings (106.2-109.0%) were found.

In vitro interactions of two pesticides, propazine and quinoxyfen with bovine serum albumin: Spectrofluorometric and molecular docking investigations.

Binding mechanisms of two selected pesticides, propazine (PRO) and quinoxyfen (QUI) with bovine serum albumin (BSA) was examined using fluorescence, absorption and molecular docking methods. Intrinsic fluorescence of BSA was quenched in the presence of both PRO and QUI. The quenching was ascertained to be conversely linked to temperature, which suggested the contribution of static quenching process in the PRO-BSA and QUI-BSA complex formations. This results were validated by the enhancement in absorption spectrum of BSA upon binding with PRO and QUI. Binding constant values (Kf = 9.55-0.60 × 10-3 M-1 for PRO-BSA system; Kf = 7.08-5.01 × 102 M-1 for QUI-BSA system) and number of binding site (n) values for the PRO-BSA and QUI-BSA systems at different temperatures affirmed a weak binding strength with a set of equivalent binding sites on BSA. Thermodynamic data obtained for both the PRO-BSA and QUI-BSA interactions predicted that the association process was spontaneous and non-covalent contacts such as hydrophobic interactions, van der Waals forces and hydrogen bonds participated in the binding reactions. This result was further supported by the molecular docking assessments. Three-dimensional spectral results revealed the microenvironmental alterations near tryptophan (Trp) and tyrosine (Tyr) residues in BSA by the addition of PRO and QUI. The docking analysis demonstrated the binding pattern for the PRO-BSA and QUI-BSA systems and disclosed the preferred binding site of both PRO and QUI as site I (subdomain IIA) of BSA.

Elucidating the interaction of antidepressant drug paroxetine with ct-dsDNA: A comparative study by electrochemical, spectroscopic, and molecular docking approaches.

This study is designed to investigate the interaction of phenylpiperidine derivative drug paroxetine, which is an effective serotonin reuptake inhibitor and biomolecules through electrochemical, fluorescence spectroscopy, and molecular docking methods. The interaction between paroxetine and biomolecules was investigated by differential pulse voltammetry according to the decrease in deoxyguanosine anodic oxidation signal of double-stranded calf thymus DNA. Fluorescence spectroscopy studies were performed by titrating paroxetine against double-stranded calf thymus DNA solution at four different temperatures. The fluorescent results showed that paroxetine had a great affinity to bind with double-stranded calf thymus DNA. Interaction studies demonstrate that paroxetine binds to double-stranded calf thymus DNA via intercalation binding mode, and the binding constant values ​​were calculated as 7.24 × 104 M-1 and 1.52 × 104 M-1 at 25 °C, based on voltammetric and spectroscopic results, respectively. Moreover, with the aim of elucidating the interaction mechanism between paroxetine and double-stranded calf thymus DNA, electrochemical and fluorescence spectroscopy studies along with molecular docking analysis were made.

Characterization of Climbazole-Bovine serum albumin interaction by experimental and in silico approaches.

Interactive association of an antifungal drug, climbazole (CBZ) with the carrier protein in bovine circulation, bovine serum albumin (BSA) was explored by fluorescence and absorption spectroscopy along with in silico techniques. The fluorescence and absorption spectral alterations of the protein upon addition of CBZ affirmed the complex foration between CBZ and BSA. The inverse temperature dependence behaviour of the KSV values as well as the hyperchromic result of the protein's absorption signals characterized CBZ-triggered quenching of BSA fluorescence as the static quenching. A weak binding affinity (Ka = 3.12-1.90-× 103 M-1) was reported towards the CBZ-BSA association process. Interpretation of thermodynamic data (entropy change = +14.68 J mol-1 K-1 and enthalpy change = -15.07 kJ mol-1) and in silico analyses anticipated that hydrophobic forces, van der Waals forces and hydrogen bonds were the key intermolecular forces in the complex stabilization. Inclusion of CBZ to BSA produced microenvironmental perturbations around Tyr and Trp residues, and also significantly defended temperature-induced destabilization of BSA. The binding locus of CBZ was detected in the proximity of Sudlow's sites I (subdomain IIA) and II (subdomain IIIA) of BSA, exhibiting greater preference towards site II, as revealed by competitive site-marker displacement investigations and in silico analysis. The stability of the CBZ-BSA complex was further validated by the molecular dynamics simulation assessments.

Electrochemical Sensor Design Based on Silver Nanoparticles Modified Multiwalled Carbon Nanotubes Paste Electrode for the Sensitive Determination of Mepivacaine.

Mepivacaine, an amide-type local anesthetic drug widely used in regional anesthesia, was studied by some aspects such as electrochemical behavior, mechanism illumination, and analytical applications by cyclic voltammetry (CV) and different pulse voltammetry (DPV) methods. In this study, a novel, fast, simple, portable, and the inexpensive electrochemical sensor was developed for the determination of mepivacaine. This study was carried out by mepivacaine anodic direction detection for the first time. The modified sensor was fabricated with silver nanoparticles (AgNP) and multiwalled carbon nanotubes paste electrode (MWCNTPE) by using the drop-dry method. Different experimental parameters, such as pulse amplitude, step potential, and scanning rate in the DPV application module, were optimized. Under optimal operation conditions, the limit of detection (LOD) as low as 31 µg L-1 was found over the dynamic range (0.1-8.0 mg L-1). In contrast to its high response towards mepivacaine, the DPV exhibits negligible responses on modified AgNP/MWCNTPE when exposed to interfering species such as dopamine, uric acid, glucose, ascorbic acid, and some heavy metals. Exceptionally, the proposed DPV method on modified AgNP/MWCNTPE was successfully applied to pharmaceutical dosage form and synthetic human serum with a low relative standard deviation (RSD) of 1.35% and 2.02%, respectively.

A Review on Electrochemical and Optical Sensing Platform Based on Ionic Liquids for Different Molecules Determination.

Since the investigation of the properties of ionic liquids, there has been an increasing interest in these materials in the field of sensors area due to their unique properties. The electrochemical and radiolytic stability of ionic liquids are prerequisites for using ionic liquids. In addition, a large number of ionic liquids can be synthesized with the combination of different anions and cations. Therefore, they are preferred to fabricate the electrochemical, optical sensors, and biosensors or used in electrolyte solutions as they promise a facile and low-cost way to examine the analytes due to their properties such as increasing the conductivity, optical properties, and stability, lowering the detection limit, facilitating the electron transfer and increasing the effective surface area of the electrode and sensor platform. All of those properties are powered with the use of nanomaterials in the modification of the sensors. In this review, general information about the properties of ionic liquids were given. Moreover, the recent developments in electrochemical and optical sensor applications in the last five years were summarized. Finally, the utilization of ionic liquids and nanomaterials in the modification of the sensor platform and the advances of the proposed sensors for the analyses taking place in different molecules were discussed.

Spectrofluorometric and computational approaches for the interaction studies of aclonifen and bifenox with human serum albumin.

Interaction of two broadly used herbicides, aclonifen (ACF) and bifenox (BIF) with the major transporter in human circulation, human serum albumin (HSA) were examined using fluorescence and absorption spectral measurements combined with in silico analyses. Assessment of the fluorescence and absorption spectral results affirmed the complexation between ACF/BIF and HSA. Increase in the KSV value with temperature characterized the ACF/BIF-induced quenching of the protein fluorescence as dynamic quenching. The moderate binding affinities (Kf = 1.74×104 - 1.95×106 M-1 for ACF-HSA complex; Kf = 2.00×103 - 1.02×106 M-1 for BIF-HSA complex) were pointed out between ACF/BIF and HSA, showing a relatively higher binding constant values with increasing temperatures. Quantitative evaluation of thermodynamic data (ΔS = +0.86 kJ mol-1 K-1 and ΔH = +225.43 kJ mol-1 for ACF-HSA complex; ΔS = +1.11 kJ mol-1 K-1 and ΔH = +304.63 kJ mol-1 for BIF-HSA complex) predicted the contribution of hydrophobic interactions in the ACF-HSA and BIF-HSA association processes, which were well supported by our molecular docking results. In silico analyses were made to acquire insight details into the ACF and BIF binding to HSA at the binding sites and suggested the locations of ACF and BIF binding sites as both subdomain IIA (site I) and subdomain IIIA (site II) of HSA, showing more preference toward site I.

An Efficient, Simultaneous Electrochemical Assay of Rosuvastatin and Ezetimibe from Human Urine and Serum Samples.

The drug combination of rosuvastatin (ROS) and ezetimibe (EZE) is used to treat hypercholesterolemia. In this work, a simultaneous electrochemical examination of ROS and EZE was conducted for the first time. The electrochemical determination of ROS and EZE was carried out using adsorptive stripping differential pulse voltammetry (AdSDPV) on a glassy carbon electrode (GCE) in 0.1 M H2SO4. The effects of the pH, scan rate, deposition potential, and time on the detection of ROS and EZE were analyzed. Under optimum conditions, the developed sensor exhibited a linear response between 1.0 × 10-6 M and 2.5 × 10-5 M for EZE and 5.0 × 10-6 M, and 1.25 × 10-5 M for ROS. The detection limits for ROS and EZE were 3.0 × 10-7 M and 2.0 × 10-6 M, respectively. The developed sensor was validated in terms of linear range, accuracy, precision, the limit of determination (LOD), and the limit of quantification (LOQ), and it was evaluated according to ICH Guidelines and USP criteria. The proposed method was also used to determine ROS and EZE in human urine and serum samples, which are reported in terms of recovery studies.

Nanomaterial-based sandwich-type electrochemical aptasensor platform for sensitive voltammetric determination of leptin.

A sandwich-type electrochemical aptasensor was designed for sensitive detection of leptin in biological samples, including human serum and human plasma. The developed aptasensor was produced by electrodeposition of gold nanoparticles on a screen-printed electrode modified with zinc oxide nanoparticles. The synergy effect of zinc oxide and gold nanoparticles improved the electrocatalytic activity of the aptasensor. The obtained high surface area allowed more aptamer molecules to be loaded on the electrode surface. Signal amplification significantly increases the detection sensitivity of a developed biosensor. Although the use of nanomaterials is the most preferred detection tool for this purpose, as an alternative, enzyme-catalyzed signal amplification is widely used in the construction of a biosensor due to its specificity and high catalytic efficiency. Therefore, both nanomaterial-supported and an alkaline phosphatase-based aptasensor design were developed, which can produce in situ electroactive product by enzymatic hydrolysis of the inactive substrate to achieve a higher signal-to-background ratio. Under optimal conditions, the developed aptasensor exhibited a wide linear concentration range from 0.01 pg mL-1 to 100.0 pg mL-1 with a detection limit of 0.0035 pg mL-1. While the developed aptasensor provided excellent selectivity in the presence of some interfering compounds, it possessed outstanding reproducibility and stability. In addition, the developed aptasensor has been applied with good recoveries in the range 96.31 to 108.79% in human serum and plasma samples. In conclusion, all the obtained results showed the feasibility of the developed aptasensor for practical applications.

Carbon-Based Composite Materials for Electrodes.

Carbon-Based Composite Materials for Electrodes is a new open Special Issue of Materials, which has the goal of publishing original research and review articles focused on carbon nanotubes, graphene, activated carbon, graphite, pencil graphite, graphene oxide, graphene nanoplatelets, pyrolytic graphite, organic mass derived carbon, fullerenes, diamond, glassy carbon, carbon fibers, and other composites for electrode preparation and its applications [...].

Evaluation of the Interaction of Cinacalcet with Calf Thymus dsDNA: Use of Electrochemical, Spectrofluorimetric, and Molecular Docking Methods.

The binding of drugs to DNA plays a critical role in new drug discovery and is important for designing better drugs. In this study, the interaction and binding mode of calf-thymus double-stranded deoxyribonucleic acid (ct-dsDNA) with cinacalcet (CIN) from the calcimimetic drug that mimics the action of calcium on tissues group were investigated. The interaction of CIN with ct-dsDNA was observed by the differential pulse voltammetry (DPV) technique by following the decrease in electrochemical oxidation signals to deoxyguanosine and adenosine. A competitive study was performed on an indicator, methylene blue, to investigate the interaction of the drug with ct-dsDNA by fluorescence spectroscopy. Interaction studies have shown that the binding mode for the interaction of CIN with ct-dsDNA could be groove-binding. According to the results obtained, the binding constant values were found to be 6.30 × 104 M-1 and 3.16 × 105 M-1, respectively, at 25 °C as obtained from the cyclic voltammetry (CV) and spectroscopic techniques. Possible molecular interactions of CIN with dsDNA were explored via molecular docking experiments. The docked structure indicated that CIN could fit well into the minor groove of the DNA through H-bonding and π-π stacking contact with CIN.

Development of the electrochemical, spectroscopic and molecular docking approaches toward the investigation of interaction between DNA and anti-leukemic drug azacytidine.

This study examines the interaction between pyrimidine nucleoside analogue azacytidine, an anti-leukemic drug, and DNA by employing electrochemical, UV-vis spectroscopy, fluorescence spectroscopy and molecular docking techniques. In the electrochemical technique, azacytidine and dsDNA interaction was investigated in two different ways: (1) in solution and (2) with a biosensor using differential pulse voltammetry (DPV) at a glassy carbon electrode. The interaction between azacytidine and dsDNA at increasing interaction times was investigated in line with the changes in adenine and guanine oxidation signals. In addition, interaction studies of polyguanine-azacytidine and polyadenine-azacytidine were performed with DPV. The binding constant values were calculated as 2.420 × 104 M-1 and 3.266 × 104 M-1 at 25 °C using UV and fluorescence spectroscopy, respectively. In conclusion, based on electrochemical and spectroscopic methods as well as molecular docking studies, it was predicted that azacytidine can bind to dsDNA via groove binding.

Lab-In-Syringe automation of deep eutectic solvent-based direct immersion single drop microextraction coupled online to high-performance liquid chromatography for the determination of fluoroquinolones.

Lab-In-Syringe direct immersion single drop microextraction is proposed as an automated sample pretreatment methodology and coupled online to HPLC with fluorescence detection for the determination of fluoroquinolones in environmental waters. For the first time, a drop of a natural deep eutectic solvent (NADES), synthesized from hexanoic acid and thymol, has been used as an extractant in automated single-drop microextraction. The extraction procedure was carried out within the 5 mL void of an automatic syringe pump. A 9-position head valve served the aspiration of all required solutions, air, waste disposal, and hyphenation with the HPLC instrument. Sample mixing during extraction was done by a magnetic stirring bar placed inside the syringe. Only 60 µL of NADES were required omitting toxic classical solvents and improving the greenness of the proposed methodology. By direct injection, linear working ranges between 0.1 and 5 µg L-1 were achieved for all fluoroquinolones. The limit of quantification values and enrichment factors ranged from 20 ng L-1 to 30 ng L-1 and 35 to 45, respectively. Accuracies obtained from the analysis of spiked surface water and wastewater treatment plant effluent analysis at two concentration levels (0.5 and 4 µg L-1) ranged from 84.6% to 119.7%, with RSD values typically <3%.

Novel Trends in Analytical Methods for ß-Blockers: An Overview of Applications in the Last Decade.

ß-blockers are among the most prescribed drugs and frequently recommended in patients with hypertension, angina, myocardial infarction, arrhythmias, and heart failure. Additionally, since ß-blockers cause low anxiety, they are abused by athletes in branches where aiming is important. ß-blocker residues may be present in environmental samples due to high consumption. At this point, the development of sensitive, rapid, and reliable analytical methods is very important for determination of ß-blockers in different matrices. Among all the analytical methods, chromatographic methods are used in a wide range for the determination of ß-blockers as they enable simultaneous determination of many analytes and removal of interfering components. Electrochemical methods, which usually do not require any pretreatment process, have become important due to their reliability, high specificity, and fast response. Simple and rapid optical methods can be used in laboratories where modern and expensive instruments such as high-performance liquid chromatography or gas chromatography are not available. The objective of this review is to provide an overview of the analytical methods for determination of ß-blockers in pharmaceuticals, biological fluids, and environmental samples. Some selected studies for the determination of ß-blockers published in the last decade are discussed. Furthermore, current trends and future perspectives on ß-blocker analyses are also suggested.

Phthalocyanine Modified Electrodes in Electrochemical Analysis.

Phthalocyanines are aromatic or macrocyclic organic compounds and attract great attention due to their numerous properties. They have many high-tech applications in different areas of the industry such as dyestuffs, thermal printing screens, photovoltaic solar cells, membrane catalytic reactors, semiconductor materials and gas sensors. In the last decade, electrochemical sensor studies have accelerated with the catalytic lighting. It plays a dominant role in the development and implementation of new generation sensors. The aim of this study is to review the electrochemical methods based on electrode modification with phthalocyanines and to shed light on new application areas of phthalocyanines. The focal point was based on the sensor applications of phthalocyanines in the determination of drugs, pesticides, organic materials and metals etc. by electrochemical methods. Experimental conditions and some validation parameters of the sensor applications such as metal phthalocyanine types, indicator electrodes, selectivity, working ranges, detection limits, and analytical applications were discussed. Consequently, this is the first review dealing with the applications of phthalocyanines in electrochemical sensors for the sensitive determination of analytes in a variety of matrices.

Analytical Methods for Determination of Antiviral Drugs in Different Matrices: Recent Advances and Trends.

Viruses are the main pathogenic substances that cause severe diseases in humans and other living things. They are among the most common microorganisms, and consequently, antiviral drugs have emerged to prevent and treat viral infections. Antiviral drugs are an essential drug group considering their prescription and consumption rates for different diseases and indications. Therefore, it is crucial to develop accurate and precise analytical methods to detect antiviral drugs in various matrices. Chromatographic techniques are used frequently for the quantification purpose since they allow simultaneous determination of antivirals. Electrochemical methods have also gained importance since the analysis can be performed quickly without the need for pretreatment. Spectrophotometric and spectrofluorimetric methods are used because they are simple, inexpensive, and less time-consuming methods. The purpose of this review is to present an overview of the analysis of currently used antiviral drugs from 2010 to 2021. Since studies on antiviral drugs are numerous, selected publications were reviewed in this article. The analysis of antiviral drugs was divided into three main groups: chromatographic, spectrometric, and electrochemical methods which were applied to different matrices, including pharmaceutical, biological, and environmental samples.

Simultaneous determination of paracetamol, dextromethorphan, chlorpheniramine, pseudoephedrine and major impurities of paracetamol and pseudoephedrine by using capillary electrophoresis

Abstract A capillary electrophoresis method was developed for the first time and optimized for the determination of paracetamol, pseudoephedrine, dextromethorphan, chlorpheniramine, 4-aminophenol and ephedrine in tablet formulation. Optimum electrophoretic conditions were achieved by using a background electrolyte of 75 mmol L-1 sodium borate buffer at pH 8.0, a capillary temperature of 30°C, a separation voltage of 30 kV and a pressure injection of the sample at 50 mbar for 10 s. Calibration graphs showed a good linearity with a coefficient of determination (R2) of at least 0.999 for all compounds. Intraday and interday precision (expressed as relative standard deviation (RSD) %) were lower than 1.39% for capillary electrophoresis method. The developed method was demonstrated to be simple and rapid for the determination of paracetamol, pseudoephedrine, dextromethorphan, chlorpheniramine, 4-aminophenol and ephedrine in tablet formulation providing recoveries in the range between 99.62 and 100.57% for all analytes.