Voltammetric determination of Salmonella typhimurium in minced beef meat using a chip-based imprinted sensor.

Early detection of pathogens is necessary for food quality monitoring, and increasing the survival rate of individuals. Conventional microbiological methods used to identify microorganisms, starting from bacterial culture and ending with advanced PCR gene identification, are time-consuming, laborious and expensive. Thus, in this study, a bacterial imprinted polymer (BIP)-based biosensor was designed and fabricated for rapid and selective detection of Salmonella typhimurium. Bio-recognition sites were made by creating template-shaped cavities in the electro-polymerized polydopamine matrices on a gold screen-printed electrode. The overall changes of the sensor, during the imprinting process, have been investigated with cyclic voltammetry, atomic force microscopy and scanning electron microscopy. The assay optimization and validation were accomplished, hence the highest sensitivity and selectivity towards S. typhimurium were achieved. As a result, a very low limit of detection of 47 CFU ml-1, and a limit of quantification of 142 CFU ml-1 were achieved using the newly-developed biosensor. No interference signals were detected when the S. typhimurium was tested in a mixed culture with other non-targeted pathogens such as Staphylococcus aureus, Listeria monocytogenes and Campylobacter jejuni. Eventually, the biosensor was applied to minced beef meat samples offering not only fast detection but also direct determination with no bacterial enrichment steps.

Recent advances in the chromatographic determination of the most commonly used anti-hepatitis C drug sofosbuvir and its co-administered drugs in human plasma.

Sofosbuvir is a direct-acting antiviral drug that inhibits hepatitis C virus (HCV) NS5B polymerase, which in turn affects the virus replication inside biological systems. The clinical importance of sofosbuvir is based not only on its effect on HCV but also on other lethal viruses such as Zika and severe acute respiratory syndrome coronavirus disease 2019 (SARS-COVID-19). Accordingly, there is a continuous shedding of light on the development and validation of accurate and fast analytical methods for the determination of sofosbuvir in different environments. This work critically reviews the recent advances in chromatographic methods for the analysis of sofosbuvir and/or its metabolites in pure samples, pharmaceutical dosage forms, and in the presence of other co-administered drugs to highlight the current status and future perspectives to enhance its determination in different matrixes.

High selectivity detection of FMDV- SAT-2 using a newly-developed electrochemical nanosensors.

Foot-and-mouth disease virus serotype South-Africa territories-2 (FMDV-SAT-2) is the most fastidious known type in Aphthovirus which is subsequently reflected in the diagnosis regime. Rapid and early diagnostic actions are usually taken in response to the FMDV outbreak to prevent the dramatic spread of the disease. Virus imprinted sensor (VIP sensor) is gathering huge attention for the selective detection of pathogens. Thus, the whole virus particles of SAT-2 together with an electropolymerized film of poly(o-phenylenediamine) (PoPD) on gold-copper modified screen-printed electrode were applied to fabricate SAT-2-virus imprinted polymer (SAT-2-VIP). The SAT-2-VIPs were fully characterized using cyclic voltammetry (CV), linear sweep voltammetry (LSV), Atomic force microscopy (AFM), Scanning electron microscope (SEM), and Fourier transform Infra-Red (FTIR) spectroscopy. Excellent selective binding affinity towards the targeted virus particle was achieved with limits of detection and quantification of 0.1 ng/mL and 0.4 ng/mL, respectively. In terms of viral interference, the sensor did not show cross-reactivity towards other animal viruses including FMDV serotype A, O, or even SAT-2 subtype Libya and the un-related virus Lumpy skin disease virus (LSDV). This high selectivity provides a sensible platform with 70 folds more sensitivity than the reference RT-PCR as revealed from the application of SAT-2-VIP sensor for rapid analysis of clinical samples with no need for treatment or equipped labs. Thus, as diagnostic and surveillance technologies, on-site point of care diagnostics for SAT-2 virus are supported.

Polyvinyl Chloride Modified Carbon Paste Electrodes for Sensitive Determination of Levofloxacin Drug in Serum, Urine, and Pharmaceutical Formulations.

Levofloxacin (LF) is a medically important antibiotic drug that is used to treat a variety of bacterial infections. In this study, three highly sensitive and selective carbon paste electrodes (CPEs) were fabricated for potentiometric determination of the LF drug: (i) CPEs filled with carbon paste (referred to as CPE); (ii) CPE coated (drop-casted) with ion-selective PVC membrane (referred to as C-CPE); (iii) CPE filled with carbon paste modified with a plasticizer (PVC/cyclohexanone) (referenced as P-CPE). The CPE was formulated from graphite (Gr, 44.0%) and reduced graphene oxide (rGO, 3.0%) as the carbon source, tricresyl phosphate (TCP, 47.0%) as the plasticizer; sodium tetrakis[3,5-bis(trifluoromethyl)phenyl] borate (St-TFPMB, 1.0%) as the ion exchanger; and levofloxacinium-tetraphenylborate (LF-TPB, 5.0%) as the lipophilic ion pair. It showed a sub-Nernstian slope of 49.3 mV decade-1 within the LF concentration range 1.0 × 10-2 M to 1.0 × 10-5 M, with a detection limit of 1.0 × 10-5 M. The PVC coated electrode (C-CPE) showed improved sensitivity (in terms of slope, equal to 50.2 mV decade-1) compared to CPEs. After the incorporation of PVC paste on the modified CPE (P-CPE), the sensitivity increased at 53.5 mV decade-1, indicating such improvement. The selectivity coefficient (log KLF2+,Fe+3pot.) against different interfering species (Na+, K+, NH4+, Ca2+, Al3+, Fe3+, Glycine, Glucose, Maltose, Lactose) were significantly improved by one to three orders of magnitudes in the case of C-CPE and P-CPE, compared to CPEs. The modification with the PVC membrane coating significantly improved the response time and solubility of the LF-TPB within the electrode matrix and increased the lifetime. The constructed sensors were successfully applied for LF determination in pharmaceutical preparation (Levoxin® 500 mg), spiked urine, and serum samples with high accuracy and precision.

Fabrication of Magnetic Molecularly Imprinted Beaded Fibers for Rosmarinic Acid.

The present study describes the fabrication of molecularly imprinted (MI) magnetic beaded fibers using electrospinning. Rosmarinic acid was selected as exemplary yet relevant template during molecular imprinting. A "design of experiments" methodology was used for optimizing the electrospinning process. Four factors, i.e., the concentration of the biodegradable polymer (polycaprolactone), the applied voltage, the flow rate, and the collector distance were varied in a central composite design. The production process was then optimized according to the suitability of the beaded fibers during microrobot fabrication, actuation, and drug release. The optimum average fiber diameter of MI beaded fibers was determined at 857 ± 390 nm with an average number of beads at 0.011 ± 0.002 per µm2. In vitro release profiles of the optimized MI beaded fibers revealed a lower burst rate and a more sustained release when compared to control fibers. Magnetic control of the MI beaded fibers was successfully tested by following selected waypoints along a star-shaped predefined trajectory. This study innovatively combines molecular imprinting technology with magnetic microrobots enabling targeted drug delivery systems that offer precise motion control via the magnetic response of microrobots along with selective uptake of a drug into the microrobot using MI beaded fibers in future.

Designing and fabrication of new VIP biosensor for the rapid and selective detection of foot-and-mouth disease virus (FMDV).

Foot and mouth disease virus (FMDV), is a highly contagious virus due to its ease of transmission. FMDV has seven genetically distinguished serotypes with many subtypes within each serotype. The traditional diagnostic methods of FMDV have demonstrated many drawbacks related to sensitivity, specificity, and cross-reactivity. In the current study, a new viral imprinted polymer (VIP)-based biosensor was designed and fabricated for the rapid and selective detection of the FMDV. The bio-recognition components were formed via electrochemical polymerization of the oxidized O-aminophenol (O-AP) film imprinted with FMDV serotype O on a gold screen-printed electrode (SPE). The overall changes in the design template have been investigated using cyclic voltammetry (CV), atomic force microscopy (AFM), Field emission-scanning electron microscopy (FE-SEM), and Fourier-transform infrared spectroscopy (FT-IR). Optimal conditions were achieved through investigating the capturing efficiency, binding stability, selectivity and life-time of the developed biosensor. The results depicted a high selectivity of the biosensor to the serotype O over all other genus serotypes A, SAT2 and Lumpy skin disease virus (LSDV), as well as, the inactivated serotype O. The limits of detection (LOD) and quantification (LOQ) were around 2 ng/mL and 6 ng/mL, respectively, in addition to the tested repeatability and reproducibility values with a variance coefficient of 1.0% and 3.6%, respectively. In comparison with the reference methods (ELISA and PCR), the analysis of saliva real samples using the developed affordable biosensor offered 50 folds lower LOD with the possibility of an on-line monitoring in the field with no prior sample treatment.

Molecularly imprinted polymers based biomimetic sensors for mosapride citrate detection in biological fluids.

Computational modeling was applied to study the intermolecular interactions in the pre-polymerization mixture and find a suitable functional monomer to use in the design of a new molecularly imprinted polymer (MIP) for mosapride citrate which is considerably a large molecule (as the citrate ion is also included in calculations as it has centers that can take part in interaction with monomer via hydrogen bonding). Based on these calculations, methacyrlic acid (MAA) was selected as a suitable functional monomer. Mosapride citrate selective MIP and a non-imprinted polymer (NIP) were synthesized and characterized using FTIR, TGA and SEM and then incorporated in carbon paste electrodes (CPEs). The designed modified sensor revealed linear responses in the ranges of 1×10-4-8×10-7 and 8×10-7-8×10-8molL-1 with a limit of detection (LOD) of 2.6×10-8molL-1. The results of the sensor exhibited high selectivity over interfering species and could be applied for the determination of mosapride citrate in pure solutions, pharmaceutical preparations, urine and human serum samples.

Preparation and application of molecularly imprinted polymer for isolation of chicoric acid from Chicorium intybus L. medicinal plant.

Molecularly imprinted polymer (MIP) was synthesized and applied for the extraction of chicoric acid from Chicory herb (Chicorium intybus L.). A computational study was developed to find a suitable template to functional monomer molar ratio for MIP preparations. The molar ratio was chosen based on the comparison of the binding energy of the complexes between the template and functional monomers. Based on the computational results, eight different polymers were prepared using chicoric acid as the template. The MIPs were synthesized in a non-covalent approach via thermal free-radical polymerization, using two different polymerization methods, bulk and suspension. Batch rebinding experiments were performed to evaluate the binding properties of the imprinted polymers. The best results were obtained with a MIP prepared using bulk polymerization with 4-vinylpyridine (4-VP) as the functional monomer and ethylene glycol dimethacrylate (EGDMA) as the crosslinker with a molar ratio of 1:4:20. The best MIP showed selective binding ability toward chicoric acid in the presence of the template's structural analogues, caffeic acid, caftaric acid and chlorogenic acid.

Synthesis and application of a molecularly imprinted polymer for the voltammetric determination of famciclovir.

A molecularly imprinted polymer (MIP) was synthesized and applied as additive within a carbon paste electrode for the cyclic voltammetric determination of famciclovir (FCV). Complementary computational studies were performed to study the intermolecular interactions in the pre-polymerization mixture. Derived from the computational studies, four MIP ratios were synthesized and their performance was evaluated using equilibrium rebinding assays. The MIP with the highest binding capacity was selected. A linear response was obtained in the range of 2.5×10(-6)-1.0×10(-3)M with a limit of detection at 7.5×10(-7)M. Finally, the developed MIP-voltammetry system was successfully applied for the determination of FCV in pure solutions and pharmaceutical preparations.

Flow injection catalase activity measurement based on gold nanoparticles/carbon nanotubes modified glassy carbon electrode.

Amperometric flow injection method of hydrogen peroxide analysis was developed based on catalase enzyme (CAT) immobilization on a glassy carbon electrode (GC) modified with electrochemically deposited gold nanoparticles on a multiwalled carbon nanotubes/chitosan film. The resulting biosensor was applied to detect hydrogen peroxide with a linear response range 1.0×10(-7)-2.5×10(-3)M with a correlation coefficient 0.998 and response time less than 10s. The optimum conditions of film deposition such as potential applied, deposition time and pH were tested and the flow injection conditions were optimized to be: flow rate of 3ml/min, sample volume 75µl and saline phosphate buffer of pH 6.89. Catalase enzyme activity was successfully determined in liver homogenate samples of rats, raised under controlled dietary plan, using a flow injection analysis system involving the developed biosensor simultaneously with spectrophotometric detection, which is the common method of enzymatic assay.