Addition Of Reagents Research Articles

Detection of Silver and Mercury Ions Using Naphthalimide-Based Fluorescent Probe

A multifunctional fluorescent probe P based on a naphthalimide derivative for the detection of Ag+ and Hg2+ through a dual-signal was designed and characterized. P exhibited a large Stokes shift (107 nm), high selectivity, good sensitivity, and fast response time. By adjusting the testing medium and the order of reagent addition, multifunctional detection with P was achieved. The addition of Ag+ or Hg2+ to P solution in either ethanol or an ethanol–water mixture resulted in a significant quenching of fluorescence emission at 537 nm and caused a decrease in the absorbance at 440 nm accompanied by the appearance of a new absorption peak at around 340 nm, and there was an obvious color change from yellow to colorless. In contrast, the addition of other common metal ions and anions did not produce substantial spectral or color changes. The detection limit of probe P for Ag+ and Hg2+ was calculated to be 0.33 μM. The sensing mechanism was proposed and validated through MS and 1H NMR spectrometry methods. Additionally, P demonstrated the capability to recognize Ag+ and Hg2+ in living cells with satisfactory results.

Highly selective split intein method for efficient separation and purification of recombinant therapeutic proteins from mammalian cell culture fluid

Biologics and vaccines have been successfully developed over the last few decades to treat many diseases. Each of these drugs must be highly purified for clinical use. Monoclonal antibodies (mAbs), the dominant therapeutic modality on the market, can be easily purified using the standard Protein A affinity platform. However, no generally applicable affinity platforms are available for the manufacture of other therapeutic proteins for clinical use. Thus, multicolumn chromatography processes for widely being used for product purification. These processes demand significant optimization to meet desired product quality attributes, where each step also decreases final yields. In this work, we demonstrate the novel self-removing iCapTagTM affinity tag, which provides a new platform for capturing, concentrating, and purifying recombinant proteins. Importantly, this system provides a tagless target protein, which is suitable for research and clinical use, where the only requirement for tag removal is a small change in buffer pH. No additional proteins, reagents or cofactors are required. We also present case studies demonstrating the use of iCapTagTM for highly efficient purification of untagged interferon alpha 2b, the ML39 single chain variable fragment (scFv), and the receptor binding domain (RBD) of SARS-CoV-2 spike protein. These proteins were expressed and secreted by Expi293 cells with the self-removing tag fused to their N-terminus. We were able to obtain highly pure (> 99%) tagless protein in a single purification step with high clearance of host cell DNA, tagged precursor, higher and lower molecular weight impurities. Based on these preliminary results, we propose the iCapTagTM as a universal capture platform for diverse classes of recombinant therapeutic proteins.

Sources, Performance and Mechanisms of Metal Ions in the Flotation Process of Copper, Lead, and Zinc Ores: A Review

In mineral processing operations, besides target minerals, slurries often contain various metal ions, including common ones with different valence states such as Pb2+, Cu2+, Fe2+, and Fe3+. These metal ions originate from multiple sources during the flotation process, including the dissolution of co-occurring metal minerals during crushing and grinding, the addition of flotation reagents, and the flotation water. Investigators have long recognized that metal ions significantly affect mineral flotation behavior. Due to physical and chemical interactions, some ions in the slurry will interact with target mineral. If these ions form hydrophobic substances on the mineral surface or increase the active sites between the mineral and collectors or sulfide agent, they will have a positive impact on the flotation process. Conversely, if they produce hydrophilic substances or deplete added collectors and sulfides, they negatively impact mineral enrichment. Meanwhile, metal ions can regulate the electrostatic repulsion between reagents and mineral surfaces in the slurry system, which has a certain impact on the flotation results. This study provides a comprehensive overview of the sources of metal ions in flotation, explores their adsorption characteristics on mineral surfaces, and examines their impact on the flotation process. It provides a theoretical basis for improving mineral flotation processes in the presence of metal ions.

Dual lateral flow test for simple and rapid simultaneous immunodetection of bisphenol A and dimethyl phthalate, two priority plastic related environmental contaminants

Contamination of the environment by technogenic endocrine disrupting compounds (EDCs) becomes serious threat to public health. To effectively prevent this threat, it is necessary to improve analytical methods for EDCs to ensure mass, fast and productive monitoring. In the given work, a dual lateral flow test (LFT) is developed in the first time for simultaneous immunodetection of bisphenol A and dimethyl phthalate, priority EDCs releasing from plastic and belonging to different chemical classes. It combines integrated detection of two EDCs by one analytical system with rapidity and simplicity of LFTs allowing for off-lab testing without additional reagents and devices. Gold nanoparticles differing in shape and color (red gold nanospheres and blue gold nanoflowers) are applied as markers to simplify interpretation of the obtained results. Under the optimal conditions chosen for efficient control of the both analytes, the detection limits of bisphenol A and dimethyl phthalate are 0.67 ng/mL and 2.22 ng/mL, respectively. Time of the assay is 15 min. The proposed dual LFT has confirmed its practical applicability by analyzing natural water samples with recovery of bisphenol A and dimethyl phthalate in the ranges of 90.4–107.0% and 86.8–118.0%, respectively.

Atomized Reagent Addition with Synchronized Jet Pre-Mineralization to Enhance the Flotation Process: Study on Atomization Parameters and Mechanisms of Enhancement

The atomized reagent and synchronous jet pre-mineralization technology, as a novel method to enhance the flotation process, increases the solubility of fatty acid collectors in pulp through atomized reagent application and improves the mineralization effect and flotation rate via synchronous jet pre-mineralization technology, thereby laying a theoretical foundation for the flotation of minerals with fatty acid collectors. Systematic studies on the atomization method, atomization particle size, and flotation experiments revealed that, compared with conventional stirring methods, the atomized reagent method increases the solubility of sodium oleate in pulp from 82.5 mg/L to 142.9 mg/L at 288.15 K. The induction time for quartz particles treated with atomized reagents and bubbles is significantly lower than that of the conventional stirring method. Semi-industrial test results of the atomized reagent and synchronous jet pre-mineralization show that, compared to traditional roughing, the TFe grade increased by 0.87 percentage points, iron recovery increased by 3.95 percentage points, and reagent consumption decreased by 7.5 percentage points. Experimental and test results demonstrate that the atomized reagent and synchronous jet pre-mineralization technology can effectively enhance mineralization, accelerate the flotation rate, improve flotation indices, and reduce reagent consumption to a certain extent, providing significant guidance for the efficient recovery of fine-grained minerals.

Stereochemical influence of 4ʹ-methyl substitutions on truncated 4ʹ-thioadenosine derivatives: Impact on A3 adenosine receptor binding and antagonism

Herein, we investigated the stereochemical effects of 4ʹ-methyl substitution on A3 adenosine receptor (A3AR) ligands by synthesizing and evaluating a series of truncated 4ʹ-thioadenosine derivatives featuring 4ʹ-α-methyl, 4ʹ-β-methyl, and 4ʹ,4ʹ-dimethyl substitutions. We successfully synthesized these derivatives, using the stereoselective addition of an organometallic reagent, KSAc-mediated sulfur cyclization, and Vorbrüggen condensation. Binding assays demonstrated that the 4ʹ-β-methyl substitution conferred the highest affinity for A3AR, with compound 1 h exhibiting a Ki = 3.5 nM, followed by the 4ʹ,4ʹ-dimethyl and 4ʹ-α-methyl substitutions. Notably, despite the absence of the 5ʹ-OH group, compound 1 h unexpectedly displayed partial agonism. Computational docking studies indicated that compound 1 h, the β-methyl derivative, adopted a South conformation and maintained strong interactions within the receptor, including a critical interaction with Thr94, a residue known to be notable for agonistic effects. Conversely, compound 2 h, the α-methyl derivative, also adopted a South conformation but resulted in a flattened structure that hindered interactions with Thr94 and Asn250. The dimethyl derivative 3 h exhibited steric clashes with Thr94, contributing to a reduction in binding affinity. However, the docking results for 3 h indicated a North conformation, suggesting that the change in sugar conformation due to the additional 4ʹ-methyl group altered the angle between the α-methyl group and the sugar plane, enabling binding despite the increased steric bulk. These findings suggest that not only do the substituents and their stereochemistry influence receptor–ligand interactions, but the conformation and the resulting spatial orientation of the substituents also play a crucial role in modulating receptor–ligand interaction. This stereochemical insight offers a valuable framework for the design of new, selective, and potent A3AR ligands, potentially facilitating the development of novel therapeutics for A3AR-related diseases such as glaucoma, inflammation, and cancer.

Green synthesis of highly fluorescent carbon quantum dots from almond resin for advanced theranostics in biomedical applications

Fluorescent Carbon Quantum Dots (CQDs) are being used in medical applications, particularly in theranostics. These Carbon Quantum Dots have been gaining more attention lately due to their potential as an effective replacement for hazardous synthetic organic dyes in a variety of biomedical applications, including live cell imaging and diagnostics. In this study, highly fluorescent Carbon Quantum Dots by one pot microwave based green route with a size of less than 10 nm, was prepared from commercially available almond resin, Prunus dulcis and conjugated with honey as additional reagent for surface functionalization. They exhibit a deep blue emission on excitation at 350 nm with an elevated quantum yield at 61%. They possess atomic nature and basic features such as high photo-stability, varying fluorescence, greater biocompatibility, and better water solubility. These fluorescent labels exhibit faster cellular invagination without disturbing the cell stability. The CQDs present cell imaging capacity with multi-coloration for visualizing the fine architecture of the nucleus naming, the nuclear membrane and nucleolus, which is linked with their varied, surface structures such as amphiphilic property and higher positive charges. These characteristics with minimal invasion have made carbon quantum dots to become the spotlight in theranostics. They can be used as alternatives to synthetic dyes for fluorescence- related cell-imaging. The intriguing fact about this approach is that it opens the possibility of combining therapy and diagnostics into one unit, which can alter how some diseases are handled and, in turn, transform the field of healthcare.

Development of a Photothermal Regenerative Plasmonic Platform as a Light-Controlled Interface.

This study introduces a novel plasmonic nanocomposite platform, where gold nanoparticles (AuNPs) are synthesized in situ within a polydimethylsiloxane (PDMS) film. The innovative fabrication process leverages ethyl acetate swelling to achieve a uniform distribution of AuNPs, eliminating the need for additional reagents. The resulting nanocomposite film exhibits exceptional photothermal conversion capabilities, efficiently converting absorbed light into heat and rapidly reaching high temperatures. Furthermore, the platform is biofunctionalized with the phosphotriesterase enzyme, not only enabling the degradation of organophosphate pesticides but also showcasing the potential for multifunctional applications. The platform's ability to be regenerated after use underscores its sustainability for repeated applications.

Simultaneous determination of somatic cell count and total plate count in raw milk based on ATP bioluminescence assay

The somatic cell count (SCC) and total plate count (TPC) are essential quality indicators for raw milk. Traditional detection methods require separate measurements and rely on complex, large-scale instruments or cultivation techniques, which are both time-consuming and laborious. To address these challenges, this study developed a novel method for the simultaneous detection of SCC and TPC in the same raw milk sample using the ATP bioluminescence assay. This method utilizes oxy-ethylated iso-nonyl phenol (Neonol-10) and cetyltrimethylammonium bromide (CTAB) to selectively lyse somatic cells and microorganisms, respectively. This technique is straightforward to operate and can be completed within 2.5 h, with detection ranges of 1 × 10⁴ to 3 × 10⁶ cells/mL for SCC and 1 × 10⁵ to 5 × 10⁷ CFU/mL for TPC. Importantly, this technique meets the requirements of detection standards in China, European Union, Canada, United States, etc. For SCC or TPC in raw milk. Overall, this innovative approach does not rely on expensive equipment or facilities and the stepwise reagent addition procedure can be easily developed into an automated high-throughput system for rapid on-site testing of SCC and TPC in raw milk.

Bioreceptors’ immobilization by hydrogen bonding interactions and differential pulse voltammetry for completely label-free electrochemical biosensors

Label-free electrochemical biosensors show great potential for the development of point-of-care devices (POCDs) for environmental and clinical applications. These sensors operate with shorter analysis times and are more economic than the labelled ones. Here, four completely label-free biosensors without electron transfer mediators were developed for hepatitis B virus (HBV) detection. The approach consisted in (i) the modification of gold surfaces with cysteamine (CT) or cysteine (CS) linkers, (ii) the subsequent antibody (Ab) immobilization, either directly by hydrogen bonding (HB) interactions or by covalent bonds (CB) using additional reagents, and (iii) measuring the biosensor response by electrochemical impedance spectroscopy (EIS) and differential pulse voltammetry (DPV). The electrode surfaces at each stage of the modification process were characterised by X-ray photo-electron spectroscopy (XPS) and atomic force microscopy (AFM). The combination of Ab immobilization by HB with the DPV analysis displayed improved repeatability, lower interference to serum matrix and similar limits of detection and quantification than the traditional biosensors that immobilize the Ab via CB and use EIS as readout technique. The Ab immobilization by HB is shown as a simple, efficient and low-cost alternative to CB ones, while DPV was faster and showed better performance than EIS. The CT-HB biosensor displayed the lowest limits of detection and quantification of 0.14 and 0.46 ng/mL, respectively, a 0.46–12.5 ng/mL linear analytical range, and 100% of recovery for 1/10 human serum media during HBV surface antigen detection by DPV. Even, it preserved the initial sensing capability after 7 days of its fabrication.Graphical abstract

A-343 Novel In-Line Hemolysis Detection on a Blood Gas Analyzer for Capillary Samples

Abstract Background Point-of-care (POC) blood gas and electrolyte analyzers utilizing whole blood as the predominant sample type are blinded by the presence of hemolysis until today. Moreover, capillary samples are particularly vulnerable to in-vitro hemolysis due to excessive force applied during sample collection. Here, we introduce the GEM Premier 7000 with iQM3 (GEM 7000) which incorporates automatic, in-line hemolysis detection in capillary blood gas samples without a need for additional sample volume or reagents. Methods The hemolysis detection module which is integrated into the disposable GEM cartridge uses acoustophoretic technology and photometric measurements to detect hemolysis in whole blood. In GEM 7000, hemolysis of ≥ 116 mg/dL was found to have significant interference with potassium results. A hemolysis flag is presented as the degree of hemolysis: none (0 - 50 and 51 - 115), mild (116 - 200 and 201 - 300), moderate (301 - 400), and gross (≥401). Due to volume limitations with native capillary samples and challenges in assessing hemolysis levels with a comparator assay, venous blood spiked volumetrically with hemolysate and transferred to capillary devices was used to verify the hemolysis flag performance. An agreement analysis was performed using the Roche cobas hemolysis value. The device's (GEM 7000) ability to flag hemolyzed capillary samples is demonstrated in a simulated clinical setting using native and contrived fingerstick capillary samples. Results Hemolysis verification with transferred capillary samples demonstrated that 98.2% (274/279) of samples were correctly flagged based on the cobas hemolysis value of spiked hemoglobin. Hemolysis in the native capillary samples (N = 235) was primarily distributed in the 0-50 mg/dL range. Additionally, native capillary samples altered using vigorous flea mixing (> 1 minute) showed hemolysis distribution over a wide hemolysis degree (none to gross), and their potassium results were flagged accordingly. Conclusions By providing integrated hemolysis detection in capillary blood sample measurements, the GEM Premier 7000 with iQM3 system can provide quality and confidence in the clinical interpretation of critical results such as potassium.

One-step green synthesis of Au@Pd concave nanocubes in deep eutectic solvents and their highly catalytic activities for selective hydrogenation

Concave nanomaterials possess superior properties, but their synthesis usually requires complicated steps, specific conditions and additional reagents (surfactants, capping agents, seeds, etc.). Herein, the well-defined Au@Pd concave nanocubes (CNCs) is facile one-step fabricated in a deep eutectic solvent (DES, named reline) composed of choline chloride (ChCl) and urea. It is found that the reline is crucial to the formation of Au@Pd CNCs. Owing to the concave core-shell structures, and electronic effects between Au core and Pd shell, Au@Pd CNCs exhibited excellent catalytic performance, and recovery convenience in the selective hydrogenation of 4-nitrostyrene. Specifically, under normal temperature and pressure conditions (25°C, 0.1 MPa), 4-nitrostyrene can be completely hydrogenated to 4-nitroethylbenzene within 7.0 min. After five cycles, Au@Pd CNCs still maintain the highly catalytic activity and 100 % selectivity toward 4-nitroethylbenzene. It is expected that well-constructed Au@Pd CNCs also display advanced potentials in other applications.

Fabrication of stable hydrogel microspheres with hydrophobic shell using water-in-water (W/W) Pickering emulsion template

Hydrogel microspheres are promising for food applications. The water-in-oil (W/O) emulsion template is commonly used to prepare the hydrogel microspheres. However, this method often involves synthetic surfactants and toxic organic solvents to remove the oil phase. The swelling problem of the resultant microspheres is another obstacle. In this study the water-in-water (W/W) emulsion template of gelatin-in-dextran was used to prepare the hydrogel microspheres, without the addition of surfactants, toxic reagents and high energy input. To prevent swelling, zein particles modified with alginate were served as a hydrophobic shell of the gelatin core. The formation evolution of these core–shell microspheres and its relevant factors including phase behavior of the immiscible gelatin/dextran, the binding of zein/alginate and the addition of the modified zein particles were investigated. It was found that small amount of alginate could induce zein particles to absorb on the gelatin surface, whereas the excess led to absorption competition. When using 0.6 wt% particles with zein/alginate ratio of 1:0.5, the ideal core–shell microspheres which were fully covered by the particles and with a self-supporting architecture in uniform size (18.8 ± 0.1 μm) and spherical shape were obtained. These microspheres showed remarkable stability under the conditions of heat treatment, varied pHs and salt concentrations and long-term storage either in refrigerator or room environment. Their sizes were easily manipulated by adjusting the concentration and molar mass of the biopolymers. It is believed that the desired stability and tunable sizes of these edible core–shell microspheres could contribute the development of their applications in foods.

Magnesium 4, 5, and 6 coordinate complexes with ligands bound via sp or sp2 hybridized atoms

As a metallic element of high natural abundance, magnesium finds a wide range of uses in both stoichiometric and more recently catalytic applications. This often takes advantage of the basic or nucleophilic properties of its compounds or their ability to co-complex with other organometallic compounds. However, the homoleptic chemistry of Mg(II) is heavily skewed towards alkyl and aryl ligands bound via sp2 and sp3 hybridized atoms. Here, we report our combined NMR spectroscopic and X-ray crystallographic study into much rarer alternative THF solvates of homoleptic magnesium complexes using ligands which bind via sp (alkynyl) and sp2 (imido) atoms. Specifically, we exploit the high acidity of terminal alkynes and diphenylacetonitrile to prepare tetra-solvated distorted octahedral complexes Mg(CCC6H4R-p)2(THF)4 (R = Me, CF3) and Mg(NCCPh2)2(THF)4 starting from the commercial alkyl reagent MgBu2. Adopting a similar deprotonative strategy using benzophenoneimine Ph2CNH affords the heteroleptic trinuclear complex Mg3(NCPh2)4nBu2(THF)2 with distorted tetrahedral Mg centres. Related imidomagnesium halide complexes can be accessed either by deprotonation of the imine with a Grignard reagent, or nucleophilic addition of a Grignard reagent to a nitrile, but these unusual five-coordinate complexes are unresponsive to a 1,4-dioxane induced Schlenk equilibrium shift. Employing a higher reflux temperature switching from a THF to a toluene medium permits access to the THF solvate of a homoleptic imido complex which also possesses a trinuclear constitution in Mg3(NCPh2)6(THF)2.

Production of Ag nanowires with high yield and narrow size distribution by promoting nucleation through hot injection and their application to disposable paper electrodes

The polyol method has been the most commonly used technique for fabricating silver nanowires (Ag NWs), wherein additive reagents facilitate precise control over their morphology. In this work, we introduced a straightforward approach to synthesize Ag NWs with minimal impurities and uniform diameter by using hot injection method. The underlying mechanism and final product are evaluated, and the scalable process was optimized to achieve a high yield (∼87.8 %). It was found that the hot injection of a Ag precursor at 170 °C significantly accelerated the reduction of Ag ions. The issue of the uniformity of Ag NWs was addressed by adding an organic halide to control the nucleation rate of Ag. The resulting Ag NWs solution was used to impregnated cellulose paper, forming an enhanced electrical network compared to other substrates. Given the low commercial cost of paper, this approach holds significant potential for widespread use in disposable electrodes. Furthermore, after subjecting the fabricated paper electrodes to various forms of damage, the connected LED bulbs consistently maintained their brightness. This demonstrates the network stability and suitability of these electrodes for flexible applications.

A Review on Some Physico-Chemical Compositions of Bread Wheat Determined by Using Destructive and Non-Destructive Methods

Some physico-chemical compositions of bread wheat (Triticum aestivum) were determined by using destructive and non-destructive methods. Standardized conventional method by Association of Official Analytical Chemists (AOAC) food analysis scheme was used for the destructive approach while near infrared spectrophotometer was used to measure some proximate compositions of wheat as a non-destructive method[1]. Near infrared techniques for obtaining the proximate compositions in the bread wheat was found to be more economical and faster without destroying the actual proximate compositions of the grains. The proximate compositions of bread wheat determined by using non-destructive measurement were moisture content 11.07 ± 0.03%, crude fiber 3.00 ± 0.00% and carbohydrate 71.80 ± 0.35% and the proximate compositions of bread wheat determined by using the destructive measurement were 9.50 ± 0.10%, 2.67 ± 0.07% and 67.28 ± 0.39%, respectively. Statistical analysis showed that there was no significant difference between the values obtained by the destructive and non-destructive methods for the determination of the physico-chemical compositions of bread wheat samples. Therefore, non-destructive method is economical, fast and save time compared to the crushing of the sample and addition of chemical reagents in the samples.

Distinguishing Organomagnesium Species in the Grignard Addition to Ketones with X‐ray Spectroscopy

The addition of Grignard reagents to ketones is a well‐established textbook reaction. However, a comprehensive understanding of its mechanism has only recently begun to emerge. X‐ray spectroscopy, because of its high selectivity and sensitivity, is the ideal tool for distinguishing between an ensemble of competing pathways. With this aim in mind, we investigated the concerted mechanism of the addition of methylmagnesium chloride (CH$_3$MgCl) to acetone in tetrahydrofuran by simulating the X‐ray spectra of different molecules in solution. We used electronic structure methods to calculate the X‐ray absorption spectra at the Mg K‐ and L$_1$‐edges and the X‐ray photoelectron spectra at the Mg K‐edge for different organomagnesium species, which coexist in solution due to the Schlenk equilibrium. The simulated spectra show that individual species can be distinguished throughout the different stages of the reaction.Each species has a distinct spectral feature which can be used as a fingerprint in solution. The absorption and photoelectron spectra consistently show a blue shift as the reaction progressed from reagents to products.

Gold nanoparticles for photothermal therapy – Influence of experimental conditions on the properties of resulting AuNPs

Gold nanoparticles (AuNPs)-mediated Photothermal Therapy (PTT) is a minimally-invasive therapeutic approach that uses AuNPs to convert light into heat, leading to the thermal ablation of tumors. Thus, the efficacy of this strategy strongly relies on the photothermal conversion potential of AuNPs. The ability to convert light into heat can be enhanced by tuning the physicochemical and optical properties of AuNPs. This can be achieved by changing the conditions of AuNP's synthesis, such as the order of addition of reagents. The present work entails to explore how varying the order of reagents addition modulates the properties of AuNPs, particularly enhancing the photothermal conversion potential of the resulting AuNPs and consequently, improving PTT efficacy. For this, eleven different AuNPs' nanoformulations were synthetized following different sequences of addition of reagents. These nanoformulations were characterized regarding their physicochemical properties namely size, surface charge, gold concentration, surface morphology and maximum absorbance wavelength. In addition, their thermal activation profiles were determined in vitro. Furthermore, the biocompatibility of different nanoformulations was also assessed. Three nanoformulations, with the most favorable photothermal activation profiles (AuNPs 2, 3 and 7), were then selected for preliminary in vitro safety and efficacy assays using a panel of cell lines. These three nanoformulations were deemed safe in vitro at the tested concentrations. At 250 μM of gold content, and after an incubation period of 4 h, followed by 5 min irradiation with a laser emitting at 808 nm (7.96 W/cm2), AuNPs 7 significantly reduced the cell viability of all cancer cell lines tested (MCF-7, HCT-116 and A375) by ≥ 45 %. However, such cytotoxic effect was not observed for the human keratinocyte cell line (HaCat), thus demonstrating its specificity towards cancer cells. Overall, the results herein presented reinforce that the order of reagents addition is highly important for achieving adequate AuNPs for PTT.

Application of validated UV-Vis spectrophotometry-colorimetric methods for specific quantification of deferiprone in the development of iron-responsive nanoparticle loaded into dissolvingmicroneedle.

Deferiprone (DFP) is one of the iron-chelating agents used in iron overload therapy for patients with ß-thalassemia major (ß-TM). However, the use of DFP is limited as it experiences a first-pass effect and can potentially cause iron deficiency due to uncontrolled release. Therefore, iron-responsive (NP-IR) DFP nanoparticle innovation was developed to control DFP release. A dissolving microneedle system (NP-IR-DMNs) was used to maximize DFP release. However, in support of this development, validation of analytical methods using spectrophotometry and colorimetrics was carried out. UV-Vis spectrophotometry is an approach that is easy to use, practical, and more cost-effective than others. The DFP levels were determined in normal and iron-overloaded medium solutions with 1%, 2%, and 4% concentrations. In addition, DFP levels were also measured in rat plasma using the colorimetric method with the addition of FeCl3 reagent to increase sensitivity for the detection of the analyte. The procedures used as guidelines in the validation procedure are The International Council for Harmonization (ICH). As a result, all linear correlation values of medium and plasma ≥ 0.999 were obtained. The LOQ levels obtained were 0.55µg/mL, 0.44µg/mL, 0.42µg/mL, 0.52µg/mL, and 1.01µg/mL in plasma, 1% FeSO4, 2% FeSO4, 4% FeSO4, and normal media, respectively. The accuracy and precision were confirmed valid, as all values were within the requirements and did not change during dilution. Then, this approach was successfully applied to determine the levels of DFP in NP-IR integrated into DMNs.

Grignard Reagent Addition to Pyridinium Salts: A Catalytic Approach to Chiral 1,4-Dihydropyridines.

Catalytic dearomatization of pyridinium salts is a powerful technique for constructing chiral N-heterocycles, which are crucial in alkaloid natural products and drugs. Despite its potential, progress in metal-catalyzed asymmetric dearomatization of pyridinium derivatives has been limited. Here, we present the enantioselective 1,4-dearomatization of pyridinium salts using Grignard reagents and chiral copper catalysis. This approach yields enantioenriched functionalized 1,4-dihydropyridines. Experimental kinetic isotope effects and density functional theory calculations provide insights into the reaction mechanism, regio- and enantioselectivity, and the rate-limiting step.