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.

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.

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.

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.

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.

A novel magnetic molecularly imprinted polymer for selective extraction and determination of quercetin in plant samples.

Molecularly imprinted polymers that mimic the binding mechanism of antibodies and their antigens exhibit several advantages, such as fast synthesis, low cost, high stability, and allow to overcome the ethical issues associated with antibody farming in animals. Herein, a novel strategy combining the magnetic molecularly imprinted polymer (MMIP) as an artificial antibody with a fluorescence procedure for the detection of quercetin in plant samples was designed. The MMIP was synthesized via a radical polymerization process to recognize specific functional groups of quercetin using a green technique based on high energy ultrasound irradiation. The developed MMIP was characterized by Fourier transform infrared spectroscopy, X-ray diffraction, scanning/transmission electron microscopy, and thermal gravimetric analysis, which confirmed the successful preparation of MMIP. The adsorption capacity and selectivity of the MMIP for quercetin and other interferents analogous were performed. The MMIP was applied in the solid-phase extraction (SPE) technique as a selective sorbent for the sample preparation. Besides, a sensitive fluorometric method for the quantitation of quercetin was developed. A linear response was obtained within the concentration of 0.005-1.25 µg mL-1 of quercetin. The limit of detection and quantitation were 1.1 ng mL-1 and 3.7 ng mL-1, respectively. The average recoveries for quercetin were between 92.2% and 104.7% with an RSD less than 5.06% in spiked orange juice and tea extract samples. Furthermore, the developed procedure was successfully combined with a new paper-based analytical device for on-site smartphone analysis of quercetin.

Development of a simplified spectrophotometric method for nitrite determination in water samples.

A new eco-friendly, rapid, and sensitive spectrophotometric method was developed to determine small quantities of nitrite, based on a diazotization mechanism. In an acidic solution, sulfathiazole was first diazotized with sodium nitrite, followed by adding phosphate buffer to form a yellow-colored compound, which showed maximum absorption at 450 nm, without the need for the addition of coupling agents such as N-(1-naphthyl) ethylenediamine. The effects of reagents amount and the optimal experimental conditions were examined by Central composite design. The simplified method presented a wide linear range of nitrite between 0.091 µg mL-1 and 1.47 µg mL-1, a sensitivity of 0.447 Abs mL µg-1, a determination coefficient of 0.998, and a low limit of detection of 0.053 µg mL-1. The simplified method was found to be comparable to the Griess method. It was evaluated for the measurements of nitrite using the accuracy profile approach. The validation procedure results established that 80% of the future results would be within the acceptability limit of 10% over the validation domain ranging from 0.174 µg mL-1 to 1.37 µg mL-1. The developed method was furtherly applied in the determination of nitrite using a developed paper-based analytical device that detected a nitrite concentration of 3 µg mL-1 which is considered by the World Health Organization to be the maximal permissible limit of nitrite in drinking water.

Preparation of Teucrium polium extract-loaded chitosan-sodium lauryl sulfate beads and chitosan-alginate films for wound dressing application.

This study aimed to formulate sodium lauryl sulfate cross-linked chitosan beads and sodium alginate-chitosan films for designing a dressing that would shorten the healing time of skin wounds. Teucrium polium extract-loaded chitosan-sodium lauryl sulfate beads (CH-SLS) and chitosan-alginate (CH-ALG) films were prepared and characterized by using Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD) analysis, and scanning electron microscopy (SEM). The swelling properties of the CH-SLS beads were also analyzed in a water solution. The obtained Teucrium polium extract-loaded CH-SLS beads and CH-ALG films (TBF) were further incorporated into the commercial adhesive dressing. This TBF wound dressing was then investigated for evaluation of its wound healing potential in the mice using the excision wound model. Healing was assessed by the macroscopic appearance and the rate of wound contraction during 8 days. On day 4, the TBF-treated wounds exhibited 98% reduction in the wound area when they were compared with healing ointment, elastic adhesive dressing, and untreated wounds which were exhibited 63%, 43%, and 32%, respectively. Furthermore, the application of TBF dressing reduced skin wound rank scores and increased the percentage of wounds contraction. These results demonstrate that TBF dressing improves considerably the healing rate and the macroscopic wound appearance at a short delay and this application may have therapeutic benefits in wound healing.