Bioinspired synergy strategy based on the integration of nanozyme into a molecularly imprinted polymer for improved enzyme catalytic mimicry and selective biosensing.

Nanozymes offer many advantages such as good stability and high catalytic activity, but their selectivity is lower than that of enzymes. This is because most of enzymes have a protein component (apoenzyme) for substrate affinity to enhance selectivity and a non-protein element (coenzyme) for catalytic activity to improve sensitivity. The synergy between molecularly imprinted polymers (MIPs) and nanozymes can mimic natural enzymes, with MIP acting as the apoenzyme and nanozyme as the coenzyme. Despite researchers' attempts to associate MIPs with nanozymes, the full potential of this combination remains not well explored. This study addresses this gap by integrating Fe3O4-Lys-Cu nanozymes with peroxidase-like catalytic activities within appropriate MIPs for L-DOPA and dopamine. The catalytic performance of the nanozyme was improved by the presence of Cu in Fe3O4-Lys-Cu and further enhanced by MIP. Indeed, the exploration of the pre-concentration property of MIP has increased twenty-fold the catalytic activity of the nanozyme. Moreover, this synergistic combination facilitated the template removal process during MIP production by reducing the extraction time from several hours to just 1 min thanks to the addition of co-substrates which trigger the reaction with nanozyme and release the template. Overall, the synergistic combination of MIPs and nanozymes offers a promising avenue for the design of artificial enzymes.

Smartphone-based device for rapid and single-step detection of piRNA-651 using anti-DNA:RNA hybrid antibody and enzymatic signal amplification.

P-element-induced wimpy testis (PIWI)-interacting RNAs (piRNAs/piRs) are a class of small noncoding RNAs that play a crucial role in regulating various biological processes, including carcinogenesis. One specific piRNA, piR-651, has been reported to be overexpressed in both human blood serum and solid cancer tissues, that can be used a viable biomarker in cancer diagnosis. Early diagnosis of cancer can help reduce the burden of the disease and improve survival rates. In the present work, we report for the first time a smartphone-based colorimetric biosensor for highly sensitive and specific detection of piR-651 thanks to an enzymatic signal amplification, which yielded high colorimetric intensities. Indeed, a heteroduplex DNA:RNA was formed in the presence of piR-651 with the capture DNA probe immobilized on the magnetic beads for easy magnetic separation. Then, a HRP tethered to anti-DNA:RNA (S9.6) was used to reveal the DNA-RNA heteroduplex formed by catalyzing the oxidation of TMB substrate into colorimetric TMBox, which absorbs at 630 nm. The absorbance is positively proportional to the piR-651 concentrations. On the other hand, the colorimetric product of the assay can be photographed with a smartphone camera and analyzed using ImageJ software. Using a smartphone and under optimal conditions, the biosensor responded linearly to the logarithm of piRNA-651 from 8 fM to 100 pM with a detection limit of 2.3 fM and discriminates against other piRNAs. It was also successfully applied to the determination of piRNA-651 levels in spiked human serum.

Molecularly imprinted photopolymers combined with smartphone-based optical sensing for selective detection of bisphenol A in foods.

Bisphenol A (BPA), known for its endocrine-disrupting properties and potential to leach into food products, has led to significant food safety concerns. Therefore, the development of sensitive and selective BPA rapid detection methods is crucial. In this study, molecularly imprinted solid-phase extraction coupled to a colorimetric method was adopted for the smartphone-based determination of BPA. The molecularly imprinted polymer (MIP) was prepared via photopolymerization and used as a selective adsorbent material for SPE columns. The solid-phase extraction (SPE) columns with multiple cycles significantly reduced the extraction time to only 30 min. The developed method demonstrates useful sensitivity for BPA (LOD = 30 ppb). Furthermore, BPA migration from plastic packaging was evaluated under different storage conditions, revealing that microwave treatment for 5 min led to BPA release from polycarbonate packaging in juice and basic solutions. The MIP selective extraction/clean-up and smartphone-based optical sensor were successfully applied to BPA standard solutions and complex food samples (e.g., juice and tap water), resulting in reproducible and selective BPA determination (RSD ≤ 6%, n = 3). This rapid and cost-effective method of producing MIPs for BPA offers a promising solution for fast and low-cost sensing for on-site fresh food analysis.

Innovative approaches to suppress non-specific adsorption in molecularly imprinted polymers for sensing applications.

Molecularly imprinted polymers (MIPs) are synthetic antibodies developed to bind selectively with specific molecules. They function through a particular recognition process involving their cavities and functional groups. Nevertheless, functional groups located outside these cavities are the main cause of non-specific molecule binding, thus reducing the effectiveness of MIPs in sensing applications. This work focused on enhancing the selectivity and performance of MIPs through electrostatic modification with surfactants. The study investigates the use of two surfactants, namely sodium dodecyl sulfate (SDS) and cetyl trimethyl ammonium bromide (CTAB), to eliminate non-specific adsorption in MIPs. The binding isotherms of the target molecule sulfamethoxazole (SMX) on MIPs and non-imprinted polymers (NIPs) were analyzed, showing higher adsorption capacity of MIPs due to the specific cavities. The modification with SDS or CTAB effectively eliminated non-specific adsorption in MIPs. The kinetic adsorption behavior further demonstrated the efficacy of MIP+--SDS/CTAB in the selective adsorption of SMX. Calibration curves showcase the methodology's analytical capabilities, achieving low limit of detection for SMX 6 ng mL-1 using MIP +-SDS. The stability study confirmed that the developed MIP +/--SDS/CTAB remains stable even at high temperatures, demonstrating its suitability for on-site applications. The methodology was successfully applied to detect SMX in milk and water samples, achieving promising recoveries. Overall, the electrostatic modification of MIPs with surfactants emerges as a valuable strategy for enhancing selectivity and performance in target molecule recognition and detection.

Selective extraction of maleic hydrazide in foods using magnetic molecularly imprinted polymers and colorimetric detection via smartphone.

Maleic hydrazide (MH) is a plant growth regulator, herbicide, and sprout inhibitor used to improve the growth and quality of certain vegetables and fruits, unfortunately, MH has genotoxic and carcinogenic effects; thus, MH residues in food need to be analyzed. Herein, magnetic molecularly imprinted polymers (MagMIP) were synthesized by radical polymerization in just 30 min using a microwave for rapid and selective extraction of MH. The colorimetric detection of MH using the immobilized Folin Ciocalteau's reagent (FCR) on 96-well microplate via smartphone sensor exhibits useful sensitivity for MH with a limit of detection (LOD = 0.6 ppm) which is far lower than the maximum residue limits (higher than 5 ppm). The immobilized FCR was stored dry at two different storage conditions at +4 °C and room temperature without losing its performance over six months. The coupling MagMIP-extraction/clean-up and smartphone determination were tested towards food samples (i.e., potatoes, and carrots), obtaining good recovery (79-96 %), high repeatability (RSD 4.5 %; n = 10), and high selectivity for MH determination.

Development of a molecularly imprinted membrane for selective, high-sensitive, and on-site detection of antibiotics in waters and drugs: Application for sulfamethoxazole.

Sulfonamides are among the widespread bacterial antibiotics. Despite this, their quick emergence constitutes a serious problem for ecosystems and human health. Therefore, there is an increased interest in developing relevant detection method for antibiotics in different matrices. In this work, a straightforward, green, and cost-effective protocol was proposed for the preparation of a selective molecularly imprinted membrane (MIM) of sulfamethoxazole (SMX), a commonly used antibiotic. Thus, cellulose acetate was used as the functional polymer, while polyethylene glycol served as a pore-former. The developed MIM was successfully characterized through scanning electron microscopy (SEM), atomic force microscopy (AFM), Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), and thermogravimetric analysis (TGA). The MIM was used as a sensing platform in conjunction with a smartphone for optical readout, enabling on-site, selective, and highly sensitive detection of SMX. In this way, a satisfactory imprinting factor of around 3.6 and a limit of detection of 2 ng mL-1 were reached after applying response surface methodologies, including Box-Behnken and central composite designs. Besides, MIM demonstrated its applicability for the accurate and selective detection of SMX in river waters, wastewater, and drugs. Additionally, the MIM was shown to be a valuable sorbent in a solid-phase extraction protocol, employing a spin column setup that offered rapid and reproducible results. Furthermore, the developed sensing platform exhibited notable regeneration properties over multiple cycles and long shelf-life in different storage conditions. The newly developed methodology is of crucial importance to overcome the limitations of classical imprinting polymers. Furthermore, the smartphone-based platform was used to surpass the typically expensive and complicated methods of detection.

Exploring the potential of molecularly imprinted polymers and metal/metal oxide nanoparticles in sensors: recent advancements and prospects.

Metal/metal oxide nanoparticles have gained increasing attention in recent years due to their outstanding features, including optical and catalytic properties, as well as their excellent conductivity. The implementation of metal/metal oxide nanoparticles, combined with molecularly imprinted polymers (MIPs) has paved the way for a new generation of building blocks to engineer and enhance the fascinating features of advanced sensors. This review critically evaluates the impact of combining metal/metal oxide nanoparticles with MIPs in sensors. It covers synthesis strategies, advantages of coupling these materials with MIPs, and addresses questions about the selectivity of these hybrid materials. In the end, the current challenges and future perspectives of this field are discussed, with a particular focus on the potential applications of these hybrid composites in the sensor field. This review highlights the exciting opportunities of using metal/metal oxide nanoparticles along with MIPs for the development of next-generation sensors.

A highly sensitive colorimetric DNA sensor for MicroRNA-155 detection: leveraging the peroxidase-like activity of copper nanoparticles in a double amplification strategy.

A novel and highly sensitive colorimetric DNA sensor for determination of miRNA-155 at attomolar levelsis presented that combines the peroxidase-like activity of copper nanoparticles (CuNPs) with the hybridization chain reaction (HCR) . The utilization of CuNPs offers advantages such as strong interaction with double-stranded DNA, excellent molecular recognition, and mimic catalytic activity. Herein, a capture probe DNA (P1) was immobilized on carboxylated magnetic beads (MBs), allowing for amplified immobilization due to the 3D surface. Subsequently, the presence of the target microRNA-155 led to the formation of a sandwich structure (P2/microRNA-155/P1/MBs) when P2 was introduced to the modified P1/MBs. The HCR reaction was then triggered by adding H1 and H2 to create a super sandwich (H1/H2)n. Following this, Cu2+ ions were attracted to the negatively charged phosphate groups of the (H1/H2)n and reduced by ascorbic acid, resulting in the formation of CuNPs, which were embedded into the grooves of the (H1/H2)n. The peroxidase-like activity of CuNPs catalyzed the oxidation reaction of 3,3',5,5'-Tetramethylbenzidine (TMB), resulting in a distinct blue color measured at 630 nm. Under optimal conditions, the colorimetric biosensor exhibited a linear response to microRNA-155 concentrations ranging from 80 to 500 aM, with a detection limit of 22 aM, and discriminate against other microRNAs. It was also successfully applied to the determination of microRNA-155 levels in spiked human serum.

Untreated vs. Treated Carbon Felt Anodes: Impacts on Power Generation in Microbial Fuel Cells.

This research sought to enhance the efficiency and biocompatibility of anodes in bioelectrochemical systems (BESs) such as microbial fuel cells (MFCs), with an aim toward large-scale, real-world applications. The study focused on the effects of acid-heat treatment and chemical modification of three-dimensional porous pristine carbon felt (CF) on power generation. Different treatments were applied to the pristine CF, including coating with carbon nanofibers (CNFs) dispersed using dodecylbenzene sulfonate (SDBS) surfactant and biopolymer chitosan (CS). These processes were expected to improve the hydrophilicity, reduce the internal resistance, and increase the electrochemically active surface area of CF anodes. A high-resolution scanning electron microscopy (HR-SEM) analysis confirmed successful CNF coating. An electrochemical analysis showed improved conductivity and charge transfer toward [Fe(CN)6]3-/4- redox probe with treated anodes. When used in an air cathode single-chamber MFC system, the untreated CF facilitated quicker electroactive biofilm growth and reached a maximum power output density of 3.4 W m-2, with an open-circuit potential of 550 mV. Despite a reduction in charge transfer resistance (Rct) with the treated CF anodes, the power densities remained unchanged. These results suggest that untreated CF anodes could be most promising for enhancing power output in BESs, offering a cost-effective solution for large-scale MFC applications.

Selective Sensing in Microbial Fuel Cell Biosensors: Insights from Toxicity-Adapted and Non-Adapted Biofilms for Pb(II) and Neomycin Sulfate Detection.

This study introduces the utilization of self-powered microbial fuel cell (MFC)-based biosensors for the detection of biotoxicity in wastewater. Current MFC-based biosensors lack specificity in distinguishing between different pollutants. To address this limitation, a novel approach is introduced, capitalizing on the adaptive capabilities of anodic biofilms. By acclimating these biofilms to specific pollutants, an enhancement in the selectivity of MFC biosensors is achieved. Notably, electrochemically active bacteria (EAB) were cultivated on 3D porous carbon felt with and without a model toxicant (target analyte), resulting in the development of toxicant-resistant anodic biofilms. The model toxicants, Pb2+ ions and the antibiotic neomycin sulfate (NS), were deployed at a concentration of 1 mg L-1 during MFC operation. The influence of toxicity on biofilm growth and power production was investigated through polarization and power density curves. Concurrently, the electrochemical activity of both non-adapted and toxicity-adapted biofilms was investigated using cyclic voltammetry. Upon maturation and attainment of peak powers, the MFC reactors were evaluated individually as self-powered biosensors for pollutant detection in fresh wastewater, employing the external resistor (ER) mode. The selected ER, corresponding to the maximum power output, was positioned between the cathode and anode of each MFC, enabling output signal tracking through a data logging system. Subsequent exposure of mature biofilm-based MFC biosensors to various concentrations of the targeted toxicants revealed that non-adapted mature biofilms generated similar current-time profiles for both toxicity models, whereas toxicity-adapted biofilms produced distinctive current-time profiles. Accordingly, these results suggested that merely by adapting the anodic biofilm to the targeted toxicity, distinct and identifiable current-time profiles can be created. Furthermore, these toxicity-adapted and non-adapted biofilms can be employed to selectively detect the pollutant via the differential measurement of electrical signals. This differentiation offers a promising avenue for selective pollutant detection. To the best of our current knowledge, this approach, which harnesses the natural adaptability of biofilms for enhanced sensor selectivity, represents a pioneering effort in the realm of MFC-based biosensing.

Fast impedimetric immunosensing of IgGs associated with peanut and hazelnut allergens.

Food allergies trigger a variety of clinical adverse symptoms and clinical evidence suggests that the presence of food allergy-related IgG can be helpful in the diagnosis when analyzed at the peptide-epitope level. To validate and select the peptides based on their specificity toward hazelnut or peanut epitopes, the authors of this study developed a silicon-based microchip coupled with click-chemistry bound peptides identified by the Fraunhofer Institute for Cell Therapy and Immunology. Peptides related to hazelnut and peanut allergies were identified and used to develop a silicon-based microchip. Peptides were coupled with click-chemistry to the sensor surface. The immunosensor was developed by electrografting diazotized amino phenylacetic acid and subsequently, dibenzocyclooctyne-amine (DBCO-NH2) was used as click-chemistry to allow coupling of the peptides with a C-terminal linker and azide structure. Energy-dispersive X-ray spectroscopy, electrochemical impedance spectroscopy (EIS), and fluorescence microscopy techniques have been used to analyze the bio-functionalization of the developed electrode. The peptide-epitope recognition was studied for seven allergen-derived peptides. The electrochemical responses were studied with sera from rabbits immunized with hazelnut and peanut powder. The microchips functionalized with the chosen peptides (peanut peptides T12 and EO13 and hazelnut peptides S4 and EO14 with an RSD of 4%, 3%, 9%, and 1% respectively) demonstrated their ability to specifically detect prevalent anti-nut related IgGs in rabbit sera in a range of dilutions from 1:500000 (0.0002%) until 1:50000 (0.002%). In addition, the other peptides showed promising differentiation abilities which can be further studied to perform multivariable detection fingerprint of anti-allergens in blood sera.

Unlocking the Potential of Molecularly Imprinted Polydopamine in Sensing Applications.

Molecularly imprinted polymers (MIPs) are synthetic receptors that mimic the specificity of biological antibody-antigen interactions. By using a "lock and key" process, MIPs selectively bind to target molecules that were used as templates during polymerization. While MIPs are typically prepared using conventional monomers, such as methacrylic acid and acrylamide, contemporary advancements have pivoted towards the functional potential of dopamine as a novel monomer. The overreaching goal of the proposed review is to fully unlock the potential of molecularly imprinted polydopamine (MIPda) within the realm of cutting-edge sensing applications. This review embarks by shedding light on the intricate tapestry of materials harnessed in the meticulous crafting of MIPda, endowing them with tailored properties. Moreover, we will cover the diverse sensing applications of MIPda, including its use in the detection of ions, small molecules, epitopes, proteins, viruses, and bacteria. In addition, the main synthesis methods of MIPda, including self-polymerization and electropolymerization, will be thoroughly examined. Finally, we will examine the challenges and drawbacks associated with this research field, as well as the prospects for future developments. In its entirety, this review stands as a resolute guiding compass, illuminating the path for researchers and connoisseurs alike.

Development of an Innovative Colorimetric DNA Biosensor Based on Sugar Measurement.

The development of biosensors for target detection plays a crucial role in advancing various fields of bioscience. This work presents the development of a genosensor that exploits the colorimetric phenol-sulfuric acid sugar reaction for the detection of DNA, and RNA as specific targets, and DNA intercalator molecules. The biosensor combines simplicity and reliability to create a novel bioassay for accurate and rapid analysis. A 96-well microplate based on a polystyrene polymer was used as the platform for an unmodified capture DNA immobilization via a silanization process and with (3-Aminopropyl) triethoxysilane (APTES). After that, a hybridization step was carried out to catch the target molecule, followed by adding phenol and sulfuric acid to quantify the amount of DNA or RNA sugar backbone. This reaction generated a yellow-orange color on the wells measured at 490 nm, which was proportional to the target concentration. Under the optimum conditions, a calibration curve was obtained for each target. The developed biosensor demonstrated high sensitivity, good selectivity, and linear response over a wide concentration range for DNA and RNA targets. Additionally, the biosensor was successfully employed for the detection of DNA intercalator agents that inhibited the hybridization of DNA complementary to the immobilized capture DNA. The developed biosensor offers a potential tool for sensitive and selective detection in various applications, including virus diagnosis, genetic analysis, pathogenic bacteria monitoring, and drug discovery.

Magnetic Molecularly Imprinted Chitosan Combined with a Paper-Based Analytical Device for the Smartphone Discrimination of Tryptophan Enantiomers.

The separation of enantiomers plays a critical role in pharmaceutical development, ensuring therapeutic efficacy, safety, and patent protection. It enables the production of enantiopure drugs and enhances our understanding of the properties of chiral compounds. In this study, a straightforward and effective chiral detection strategy was developed for distinguishing between tryptophan (TRP) enantiomers. The approach involved the preparation of a magnetic molecularly imprinted chitosan (MMIC) for preparation of the sample, which was combined with a nitrocellulose membrane (a paper-based analytical device, PAD) integrated with D-TRP covalently grafted with polymethacrylic acid (PAD-PMA_D-TRP). Discriminating between the TRP enantiomers was achieved using AuNPs as a colorimetric probe. Indeed, the presence of D-TRP rapidly induced the aggregation of AuNPs due to its strong affinity to PAD-PMA_D-TRP, resulting in a noticeable change in the color of the AuNPs from red to purple. On the other hand, L-TRP did not induce any color changes. The chiral analysis could be easily performed with the naked eye and/or a smartphone. The developed method exhibited a detection limit of 3.3 µM, and it was successfully applied to detect TRP in serum samples, demonstrating good recovery rates. The proposed procedure is characterized by its simplicity, cost-effectiveness, rapidity, and ease of operation.

Feather-like Gold Nanostructures Anchored onto 3D Mesoporous Laser-Scribed Graphene: A Highly Sensitive Platform for Enzymeless Glucose Electrochemical Detection in Neutral Media.

The authors present a novel sensing platform for a disposable electrochemical, non-enzymatic glucose sensor strip at physiological pH. The sensing material is based on dendritic gold nanostructures (AuNs) resembling feather branches, which are electrodeposited onto a laser-scribed 3D graphene electrode (LSGE). The LSGEs were fabricated via a one-step laser scribing process on a commercially available polyimide sheet. This study investigates several parameters that influence the morphology of the deposited Au nanostructures and the catalytic activity toward glucose electro-oxidation. The electrocatalytic activity of the AuNs-LSGE was evaluated using cyclic voltammetry (CV), linear sweep voltammetry (LSV), and amperometry and was compared to commercially available carbon electrodes prepared under the same electrodeposition conditions. The sensor demonstrated good stability and high selectivity of the amperometric response in the presence of interfering agents, such as ascorbic acid, when a Nafion membrane was applied over the electrode surface. The proposed sensing strategy offers a wide linear detection range, from 0.5 to 20 mM, which covers normal and elevated levels of glucose in the blood, with a detection limit of 0.21 mM. The AuNs-LSGE platform exhibits great potential for use as a disposable glucose sensor strip for point-of-care applications, including self-monitoring and food management. Its non-enzymatic features reduce dependence on enzymes, making it suitable for practical and cost-effective biosensing solutions.

Immobilization of diphenylcarbazide on paper-based analytical devices for the pre-concentration and detection of chromium VI in water samples.

A novel approach using a smartphone for the detection of Cr (VI) has been developed. In this context, two different platforms were designed for the detection of Cr (VI). The first one was synthesized via a crosslinking reaction of chitosan with 1,5-Diphenylcarbazide (DPC-CS). The obtained material was integrated into a paper to develop a new paper-based analytical device (DPC-CS-PAD). The DPC-CS-PAD exhibited high specificity toward Cr (VI). The second platform (DPC-Nylon PAD) was prepared by covalent immobilization of DPC onto a Nylon paper and then its analytical performances regarding Cr (VI) extraction and detection were evaluated. DPC-CS-PAD presented a linear range of 0.1-5 ppm with detection and quantification limits of about 0.04 and 0.12 ppm, respectively. The DPC-Nylon-PAD exhibited a linear response of 0.1-2.5 ppm with detection and quantification limits of 0.06 and 0.2 ppm, respectively. Furthermore, the developed platforms were effectively applied for testing the effect of the loading solution volume for trace Cr (IV) detection. For the DPC-CS material, a volume of 20 mL allowed the detection of 4 ppb of Cr (VI). In the case of DPC-Nylon-PAD, the loading volume of 1 mL permitted the detection of the critical concentration of Cr (VI) in water.

Geographical Classification of Saffron (Crocus Sativus L.) Using Total and Synchronous Fluorescence Combined with Chemometric Approaches.

There is an increasing interest in food science for high-quality natural products with a distinct geographical origin, such as saffron. In this work, the excitation-emission matrix (EEM) and synchronous fluorescence were used for the first time to geographically discriminate between Moroccan saffron from Taroudant, Ouarzazate, and Azilal. Moreover, to differentiate between Afghan, Iranian, and Moroccan saffron, a unique fingerprint was assigned to each sample by visualizing the EEM physiognomy. Moreover, principal component analysis (LDA) and linear discriminant analysis (LDA) were successfully applied to classify the synchronous spectra of samples. High fluorescence intensities were registered for Ouarzazate and Taroudant saffron. Yet, the Azilal saffron was distinguished by its low intensities. Furthermore, Moroccan, Afghan, and Iranian saffron were correctly assigned to their origins using PCA and LDA for different offsets (Δλ) (20-250 nm) such that the difference in the fluorescence composition of the three countries' saffron was registered in the following excitation/emission ranges: 250-325 nm/300-480 nm and 360-425 nm/500-550 nm. These regions are characterized by the high polyphenolic content of Moroccan saffron and the important composition of Afghan saffron, including vitamins and terpenoids. However, weak intensities of these compounds were found in Iranian saffron. Furthermore, a substantial explained variance (97-100% for PC1 and PC2) and an important classification rate (70-90%) were achieved. Thus, the non-destructive applied methodology of discrimination was rapid, straightforward, reliable, and accurate.

Development of a New Route for the Immobilization of Unmodified Single-Stranded DNA on Chitosan Beads and Detection of Released Guanine after Hydrolysis.

In this work, chitosan beads were used as a cost-effective platform for the covalent immobilization of unmodified single-stranded DNA, using glutaraldehyde as a cross-linking agent. The immobilized DNA capture probe was hybridized in the presence of miRNA-222 as a complementary sequence. The target was evaluated based on the electrochemical response of the released guanine, using hydrochloride acid as a hydrolysis agent. Differential pulse voltammetry technique and screen-printed electrodes modified with COOH-functionalized carbon black were used to monitor the released guanine response before and after hybridization. The functionalized carbon black provided an important signal amplification of guanine compared to the other studied nanomaterials. Under optimal conditions (6 M HCl at 65 °C for 90 min), an electrochemical-based label-free genosensor assay exhibited a linear range between 1 nM and 1 µM of miRNA-222, with a detection limit of 0.2 nM of miRNA-222. The developed sensor was successfully used to quantify miRNA-222 in a human serum sample.

Paper-Based Colorimetric Detection of miRNA-21 Using Pre-Activated Nylon Membrane and Peroxidase-Mimetic Activity of Cysteamine-Capped Gold Nanoparticles.

Irregular expression of MicroRNA-21 (miRNA-21) is considered as a promising biomarker for early cancer diagnosis. In this paper, a new genosensor based on paper and nanozyme activity of cysteamine-capped gold nanoparticles (Cys/AuNPs) was developed to detect picomolar concentrations of miRNA-21. Such nanozyme catalyzes the colorimetric reaction of hydrogen peroxide (H2O2) and 3,3',5,5' tetramethylbenzidine (TMB), to produce a blue color measurable by a smartphone. Due to their positive charge, Cys/AuNPs were attached to the negative phosphate groups of the DNA strand backbone via electrostatic interactions, leading to the quantitative determination of miRNA-21 concentration by the peroxidase-like activity of Cys/AuNPs. Furthermore, a paper-based assay was carried out on nylon disk devices to allow fast immobilization of DNAprobe. After performing the paper-based assay, a good linear range was observed between 1 pM and 1 nM (Y = 0.080 [MiRNA-21]/pM + 13.846, R2 = 0.993) with a detection limit of 0.5 pM. The developed method was effective, selective, and sensitive for the miRNA-21 detection. The application of the proposed method for miRNA-21 detection was examined in a human serum sample, and a recovery rate of 90.0-97.6% was obtained showing the acceptable accuracy of the developed approach.

Study on Molecularly Imprinted Polymers Obtained Sonochemically for the Determination of Aflatoxins in Food.

Aflatoxins (AFs) are fungi secondary metabolites produced by the Aspergillus family. These compounds can enter the food chain through food contamination, representing a risk to human health. Commercial immunoaffinity columns are widely used for the extraction and cleanup of AFs from food samples; however, their high cost and large solvent consumption create a need for alternative strategies. In this work, an alternative strategy for producing molecularly imprinted polymers (MIPs) was proposed to extract aflatoxins AFB1, AFB2, AFG1, and AFG2 from complex food samples, using liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). The MIPs were synthesized via a low-cost and rapid (5 min) sonochemical free-radical polymerization, using 1-hydroxy-2-naphthoic acid as a dummy template. MIPs-based solid phase extraction performance was tested on 17 dietary supplements (vegetables, fruits, and cereals), obtaining appreciable recovery rates (65-90%) and good reproducibility (RSD ≤ 6%, n = 3); the selectivity towards other mycotoxins was proved and the data obtained compared with commercial immunoaffinity columns. The proposed strategy can be considered an alternative affordable approach to the classical immunoaffinity columns, since it is more selective and better performing.