Aqueous Buffer pH Research Articles

A water-soluble colorimetric and turn-on fluorescence sensor for fast and sensitive detection of sulfite/bisulfite ions in real samples and live cells

Consuming an excessive amount of sulfites can have detrimental effects on human health. Therefore, it is essential to develop an effective technique to detect the presence of HSO3− in food samples and live cells. Herein, we have developed a colorimetric and turn-on fluorescent probe PI capable of efficiently detecting sulfite/ bisulfite (SO32− /HSO3−) in aqueous PBS buffer (pH 7.4). This probe PI is non-fluorescent due to intramolecular charge transfer (ICT), but becomes fluorescent in the presence of HSO3−. The reaction between SO32− /HSO3− and the probe disrupts π - conjugation by a nucleophilic addition mechanism, resulting in the inhibition of intramolecular charge transfer (ICT). This leads to changes in both the fluorescence and absorption spectra. The color of the probe PI changed from blue to colorless, making it easier to detect HSO3−with the naked eye. Our probe also demonstrates excellent sensitivity, selectivity, fast (reaction completed in 300 s), and a low limit of detection (LOD = 8.4 nM). Furthermore, it is effective in detecting levels of SO32− /HSO3− in water, sugar, and MDA-MB-231 live cancer cells.

A new process related impurity identification, characterization, development of analytical method by HPLC and method validation for Voriconazole API

AbstractDuring the testing of laboratory Voriconazole API batches, one unidentified impurity (IMP-5.312) was detected employing the Pharmeuropa HPLC technique at a level in excess of 0.10%. This IMP-5.312 was synthesized and then characterized as 6-(3-(2,4-difluorophenyl)-3-hydroxy-4-(1H-1,2,4-triazol-1-yl) butan-2-yl)-5-fluoropyrimidin-4-ol by the corresponding spectral information (MS, 1H-NMR, 13C-NMR, and IR). The IMP-5.312 impurity was effectively quantified using an enhanced HPLC based-technique that was developed as well as validated. The approach made use of a Novapak C18 column with an inner diameter of 3.9 mm and a length of 150 mm (4.0 µm) for chromatographic separation. The analysis of IMP-5.312 was made at 45 °C, with a flow rate (isocratic) of 1.0 mL min−1 and a 256 nm detection wavelength. Acetonitrile, methanol, and 0.1% aqueous trifluoro acetate buffer (pH 4.0) were mixed at a ratio of 15:30:55 (v/v/v) to create the mobile phase for a 20 μL sample injection. The linearity range of 0.25281–1.51690 μg mL−1 had a correlation coefficient more than 0.99942, and the accuracy ranged from 89.3 to 100.3%. It was noted that the established HPLC based-technique was sensitive, specific, and precise. The technique was executed on the current batches of VRC API for IMP-5.312 analysis, and the outcomes were good. For quality control purposes during the manufacturing procedure of VRC, the identification as well as analysis of IMP-5.312 should be helpful. The in silico approach was applied to predict the IMP-5.312 toxicity. The reports indicated that IMP-5.312 in non-mutagenic and categorized as ICH M7 class-5 impurity.

Utilizing central composite design (CCD) in the optimization of new HPLC-PDA method for the assay of daridorexant: Application to the analysis in bulk and tablet dosage forms

In this study, we developed a robust Quality by Design guided reverse phase high-performance liquid chromatographic method for quantifying daridorexant with simplicity, speed, sensitivity, reproducibility, and cost-effectiveness. Utilizing a Waters e2690 alliance HPLC system with Empower 2® software control and a SpursilTM C18 HPLC column, critical method parameters such as flow rate, % aqueous buffer and buffer pH were identified through preliminary trials and risk assessment studies. Optimization via central composite design as part of a Design of Experiments framework, with retention time and tailing factor as critical analytical attributes, resulted in optimized chromatographic conditions: 30:70 (% v/v) phosphate buffer (pH 3) and acetonitrile as mobile phase, flowing at 1.0 mL/min, with detection at 270 nm. The method exhibited linearity within a range of 2.0-10.0 μg/ mL (correlation coefficient: 0.9997) and detection, quantitation limits of 0.089 μg/mL and 0.271 μg/mL, respectively. Validation according to ICH Q2 (R1) guidelines confirmed the method’s robustness, precision, accuracy, sensitivity, and high linearity for daridorexant estimation in both bulk drug and marketed formulations.

Synthesis and characterization of a unique fused spiro-dihydropyran based chemodosimeter for selective visual and fluorescent detection of Cu2+ ion in aqueous medium by “turn-on” mode

A realistically newly designed Cu2+ ion specific fluorescent chemodosimeter has been synthesized from α,α'-(E,E)-bis(benzylidene)cyclohexanone having a fused spiro-dihydropyran moiety characterized by 2D NMR and X-Ray Crystallographic study. It also demonstrates an irreversible, instant direct visualisation by unaided eye chemosensing property in aqueous buffer (pH 7.4) with a colour change from yellow to orange upto 0.3 × 10−6 M.

Development and Validation of RP-HPLC Method using UV detection for Simultaneous quantification of Amlodipine besylate and Nebivolol hydrochloride in Fixed-dose combination tablets

Calcium channel blockers and beta blockers are the class of choice of drugs in treatment of hypertension and cardio vascular diseases. Such a fixed dose combination is viable to deal with the patients for whom monotherapy is not sufficient. A simple, responsive and productive high performance liquid chromatography method was designed and approved for simultaneous quantification of Amlodipine Besylate (AMB) and Nebivolol Hydrochloride (NBL) in tablet dosage form. In the developed method mobile phase comprised of a mixture of aqueous buffer (pH 6) and acetonitrile (55:45 v/v, respectively) at a flow rate of 1.5 ml/min. Chromatographic separation was achieved using Reverse-phase Kromasil 60-5-CN column, 250 × 4.6 mm (particle size 5 μm). Method was discovered specific as no obstruction of excipients peaks at the retention time of tablets was observed and demonstrated linearity in concentration range of 30.4-70.8 mcg/ml for AMB and 29.8-69.4 mcg/ml for NBL with R2> 0.999. Accuracy values for AMB and NBL was found to be 98.21-99.49% and 97.91-99.09%, respectively. Inter-day and intra-day precision was carried out and % RSD was found to be less than 2%. Robustness was performed to evaluate the results with slight variations in method and % RSD found to be less than 2%. Solution stability studies were conducted in normal conditions. Force degradation study was carried out using Photo-diode Array (PDA) detector and revealed that acid degradation in AMB and NBL was 7.12% and 1.88%, alkaline degradation was 9.61% and 0.05% and oxidative degradation was 8.50% and 1.59%, respectively.

Photophysical isomerization of liposomal bilayer-included caffeic acid phenethyl ester leading to membrane dehydration

Caffeic acid phenethyl ester (CAPE) is well known for its prolific biological and pharmaceutical applications. There are plethora of studies on CAPE in context to antimicrobial, antiviral, anti-inflammatory and anticancer activities. It has been applied even to suppress chemotherapy-induced toxicities. However, an important property of CAPE has been so far overlooked, which is its photoinduced cis/trans isomerization. The present study has enlightened upon the detailed photophysical changes and the related physical parameters in CAPE. CAPE, being a membrane binding molecule, has been added to the hydrophobic portion of the bilayer of large unilamelar vesicles (LUVs) made from dimyristoyltrimethylammonium propane (DMTAP) lipids in aqueous buffer (pH 7.4). Absorption of light and conversion of the trans-isomers to the cis-form led to vesicular squeezing followed by expulsion of the penetrated water molecules that hydrated the liposomal membrane.

Role of Multiple Vanadium Centers on Redox Buffering and Rates of Polyvanadomolybdate-Cu(II)-Catalyzed Aerobic Oxidations.

A recent report established that the tetrabutylammonium (TBA) salt of hexavanadopolymolybdate TBA4H5[PMo6V6O40] (PV6Mo6) serves as the redox buffer with Cu(II) as a co-catalyst for aerobic deodorization of thiols in acetonitrile. Here, we document the profound impact of vanadium atom number (x = 0-4 and 6) in TBA salts of PVxMo12-xO40(3+x)- (PVMo) on this multicomponent catalytic system. The PVMo cyclic voltammetric peaks from 0 to -2000 mV vs Fc/Fc+ under catalytic conditions (acetonitrile, ambient T) are assigned and clarify that the redox buffering capability of the PVMo/Cu catalytic system derives from the number of steps, the number of electrons transferred each step, and the potential ranges of each step. All PVMo are reduced by varying numbers of electrons, from 1 to 6, in different reaction conditions. Significantly, PVMo with x ≤ 3 not only has much lower activity than when x > 3 (for example, the turnover frequencies (TOF) of PV3Mo9 and PV4Mo8 are 8.9 and 48 s-1, respectively) but also, unlike the latter, cannot maintain steady reduction states when the Mo atoms in these polyoxometalate (POMs) are also reduced. Stopped-flow kinetics measurements reveal that Mo atoms in Keggin PVMo exhibit much slower electron transfer rates than V atoms. There are two kinetic arguments: (a) In acetonitrile, the first formal potential of PMo12 is more positive than that of PVMo11 (-236 and -405 mV vs Fc/Fc+); however, the initial reduction rates are 1.06 × 10-4 s-1 and 0.036 s-1 for PMo12 and PVMo11, respectively. (b) In aqueous sulfate buffer (pH = 2), a two-step kinetics is observed for PVMo11 and PV2Mo10, where the first and second steps are assigned to reduction of the V and Mo centers, respectively. Since fast and reversible electron transfers are key for the redox buffering behavior, the slower electron transfer kinetics of Mo preclude these centers functioning in redox buffering that maintains the solution potential. We conclude that PVMo with more vanadium atoms allows the POM to undergo more and faster redox changes, which enables the POM to function as a redox buffer dictating far higher catalytic activity.

QCM‐D Study of the Formation of Solid‐Supported Artificial Lipid Membranes: State‐of‐the‐Art, Recent Advances, and Perspectives

Solid‐supported lipid bilayers (SLBs) are excellent platforms for studying the biophysical properties of cell membranes. Among the existing approaches used to form SLBs, vesicle fusion and rupture onto solid supports are most commonly employed owing to their straightforward procedure. The current understanding of the mechanisms behind this approach has greatly benefited from the use of surface‐sensitive techniques, especially quartz crystal microbalance with dissipation monitoring (QCM‐D) in combination with other analytical techniques, such as atomic force microscopy (AFM) or localized surface plasmon resonance (LSPR). Herein, an overview of the pathways of vesicle adsorption and rupture under various experimental conditions is provided. Examples including recent findings of how the variation of the properties of lipid vesicles (size, charge), aqueous buffer (pH, ionic strength, osmotic pressure), and solid support (surface energy) affect the pathway mechanism of adsorption and rupture are provided. Recent reports on poorly understood properties such as surface roughness and topography are provided, together with the need for further studies relevant to biomimetic and sensing purposes.

Thioester deprotection using a biomimetic NCL approach.

The reversibility of the thiol-thioester linkage has been broadly employed in many fields of biochemistry (lipid synthesis) and chemistry (dynamic combinatorial chemistry and material science). When the transthioesterification is followed by a S-to-N acyl transfer to give an amide bond, it is called Native Chemical Ligation (NCL), a high-yield chemoselective process used for peptide synthesis. Recently, we described thioglycolic acid (TGA) as a useful reagent for thioester deprotection both in solution and anchored to a solid-support under mild conditions. Inspired by NCL, in this work, we extended this approach and explored the use of 2-aminothiols for the deprotection of thiols bearing an acyl group. The best results were obtained using cysteamine or L-cysteine in an aqueous buffer pH 8 at room temperature for 30 min. The described approach was useful for S-acetyl, S-butyryl, and S-benzoyl heterocycles deprotection with yields up to 84%. Employing this methodology, we prepared six new analogs 2 of mercaptomethyl bisthiazolidine 1, a useful inhibitor of a wide-range of Metallo-β-Lactamases (MBLs). Compared with the previous methodologies (TGA polymer supported and TGA in solution), the biomimetic deprotection herein described presents better performance with higher yields, shorter reaction times, less time-consuming operations, easier setup, and lower costs.

Investigation of Factors Influencing Formation of Nanoemulsion by Spontaneous Emulsification: Impact on Droplet Size, Polydispersity Index, and Stability.

Interest in nanoemulsion technology has increased steadily in recent years for its widespread applications in the delivery of pharmaceuticals, nutraceuticals, and cosmeceuticals. Rational selection of the composition and the preparation method is crucial for developing a stable nanoemulsion system with desired physicochemical characteristics. In the present study, we investigate the influence of intricate factors including composition and preparation conditions that affect characteristic parameters and the stability of the nanoemulsion formation prepared by the spontaneous emulsification method. Octanoic acid, capryol 90, and ethyl oleate were selected to represent oil phases of different carbon–chain lengths. We explored the impact of the addition mode of the oil–Smix phase and aqueous phase, vortexing time, Km (surfactant/cosurfactant) ratio, and the replacement of water by buffers of different pH as an aqueous system. The phase behavior study showed that the Smix phase had a significant impact on the nanoemulsifying ability of the nanoemulsions composed of oil phases of varying carbon-chain lengths. The mode of mixing of the oil–Smix phase to the aqueous phase markedly influenced the mean droplet size and size distribution of the nanoemulsions composed of oil phases as capryol 90. Vortexing time also impacted the mean droplet size and the stability of the generated nanoemulsion system depending on the varying carbon-chain length of the oil phase. The replacement of the water phase by aqueous buffers of pH 1.2, 5.5, 6.8, and 7.4 has altered the mean droplet size and size distribution of the nanoemulsion system. Further, the Km ratio also had a significant influence on the formation of the nanoemulsion system. The findings of this investigation are useful in understanding how the formulation composition and process parameters of the spontaneous emulsification technique are responsible for affecting the physicochemical characteristics and stability of the nanoemulsion system composed of oil of varying carbon-chain (C8-C18) length.

Reagent-Free Immobilization of Industrial Lipases to Develop Lipolytic Membranes with Self-Cleaning Surfaces.

Biocatalytic membrane reactors combine the highly efficient biotransformation capability of enzymes with the selective filtration performance of membrane filters. Common strategies to immobilize enzymes on polymeric membranes are based on chemical coupling reactions. Still, they are associated with drawbacks such as long reaction times, high costs, and the use of potentially toxic or hazardous reagents. In this study, a reagent-free immobilization method based on electron beam irradiation was investigated, which allows much faster, cleaner, and cheaper fabrication of enzyme membrane reactors. Two industrial lipase enzymes were coupled onto a polyvinylidene fluoride (PVDF) flat sheet membrane to create self-cleaning surfaces. The response surface methodology (RSM) in the design-of-experiments approach was applied to investigate the effects of three numerical factors on enzyme activity, yielding a maximum activity of 823 ± 118 U m−2 (enzyme concentration: 8.4 g L−1, impregnation time: 5 min, irradiation dose: 80 kGy). The lipolytic membranes were used in fouling tests with olive oil (1 g L−1 in 2 mM sodium dodecyl sulfate), resulting in 100% regeneration of filtration performance after 3 h of self-cleaning in an aqueous buffer (pH 8, 37 °C). Reusability with three consecutive cycles demonstrates regeneration of 95%. Comprehensive membrane characterization was performed by determining enzyme kinetic parameters, permeance monitoring, X-ray photoelectron spectroscopy, FTIR spectroscopy, scanning electron microscopy, and zeta potential, as well as water contact angle measurements.

Cation Bulk and pKa Modulate Diblock Polymer Micelle Binding to pDNA.

Polymer-based gene delivery relies on the binding, protection, and final release of nucleic acid cargo using polycations. Engineering polymeric vectors, by exploring novel topologies and cationic moieties, is a promising avenue to improve their performance, which hinges on the development of simple synthetic methods that allow facile preparation. In this work, we focus on cationic micelles formed from block polymers, which are examined as promising gene compaction agents and carriers. In this study, we report the synthesis and assembly of six amphiphilic poly(n-butyl acrylate)-b-poly(cationic acrylamide) diblock polymers with different types of cationic groups ((dialkyl)amine, morpholine, or imidazole) in their hydrophilic corona. The polycations were obtained through the parallel postpolymerization modification of a poly(n-butyl acrylate)-b-poly(pentafluorophenyl acrylate) reactive scaffold, which granted diblock polymers with equivalent degrees of polymerization and subsequent quantitative functionalization with cations of different pKa. Ultrasound-assisted direct dissolution of the polycations in different aqueous buffers (pH = 1-7) afforded micellar structures with low size dispersities and hydrodynamic radii below 100 nm. The formation and properties of micelle-DNA complexes ("micelleplexes") were explored via DLS, zeta potential, and dye-exclusion assays revealing that binding is influenced by the cation type present in the micelle corona where bulkiness and pKa are the drivers of micelleplex formation. Combining parallel synthesis strategies with simple direct dissolution formulation opens opportunities to optimize and expand the range of micelle delivery vehicles available by facile tuning of the composition of the cationic micelle corona.

Synthesis, selective decoration and photocrosslinking of self‐immolative poly(thioester)‐PEG hydrogels

AbstractSelf‐immolative polymers are a class of polymers that undergo linear depolymerization of the main chain, often triggered by a single initial degradation event. Chain‐unzipping immolation processes followed by low‐ceiling‐temperature polymers yield regenerated monomers, demonstrating the remarkable potential of these polymers in the development of reversible and recyclable materials. In this work, we describe the synthesis of an amino‐acid‐derived poly(thioester) self‐immolative polymer with pendent functional groups allowing for selective decoration of the polymer backbone with solubilizing and reactive functional groups for subsequent post‐polymerization modification. Crosslinking norbornene‐functionalized, water‐soluble derivatives of these polymers via the thiol‐ene photoclick reaction yielded mechanically robust hydrogels within 15 s of light exposure. These networks were capable of immolation in response to deprotonation of an end‐capping thiol, unzipping along the poly(thioester) backbone to yield constituent monomers and larger molecular weight crosslinker fragments. Finally, degelation rates were controlled with pH and temperature in aqueous buffers. At physiological pH and temperature, complete degradation of approximately 20 wt% hydrogels was observed after 16 h, and raising the buffer pH to 10 resulted in network dissolution within 5 h. © 2022 Society of Industrial Chemistry.

Solubility/distribution thermodynamics and permeability of two anthelmintics in biologically relevant solvents

The solubility of mebendazole and flubendazole was measured in aqueous buffers (pH 2.0 and pH 7.4), hexane, and octanol in the temperature range of 293.15–313.15 K by the shake flask method. The molecule ionization state effect on the solubility was investigated. The ionization constants of the compounds at different temperatures were derived from the solubility data. The distribution coefficients in the octanol/buffer (pH 2.0) system at (293.15–313.15 K) and in the hexane/buffer (pH 2.0) system at 298.15 K were measured. The hydrogen bonding potential of the compounds was evaluated using the ΔlogD parameter. The thermodynamic functions of the dissolution and transfer process between the organic and aqueous phases were calculated and discussed. The permeability coefficients through the PermeaPad barrier were determined. The dependences between the distribution and permeability parameters and structural descriptor ΣCad/α were analyzed to obtain the correlation equations.

Separation of fosetyl and phosphonic acid in food matrices with mixed-mode HPLC column coupled with tandem mass spectrometric detection and method application to other highly polar pesticides

The aluminum salt of fosetyl (tris(ethyl phosphonate)) is an antifungal agrochemical. This paper presents a novel high performance liquid chromatography (HPLC) method for the simultaneous determination of fosetyl and the phosphonic acid, its main metabolite, in food samples. The method is based on an ion-displacement separation performed on the recently released Luna Omega PS C18 mixed-mode HPLC column. Baseline separation of fosetyl and phosphonic acid was feasible. This was achieved by optimizing the mobile phase composition and by introducing ethylenediaminetetraacetate for all matrices in the generally used extraction medium for polar pesticides and the injection solution. The binary mobile phase consisted of 10% (v/v) methanol in water and aqueous formate buffer (pH = 3.5) in gradient elution mode. The main advantages of the method over previous method include the stable retention time and peak resolution without the need for long column priming, conditioning or regeneration. Moreover, the approach was tested with other polar pesticides including glyphosate, glufosinate, and perchlorate and showed fit-for-purpose separation. The method was validated for spinach, cherry, and wheat flour samples, and was successfully applied on oat flour and arugula quality control samples. The results obtained for the five analytes met the requirements set by EU. The limit of quantifications was much lower than the maximum residue limits and ranged from 0.02 to 0.20 mg/kg.

Synthesis of a new environment-sensitive fluorescent probe based on TICT and application for detection of human serum albumin and specific lipid droplets imaging

Environment-sensitive fluorescent probes have always been as forceful tools to understand the pathophysiological processes of relevant diseases. In this work, a new fluorescent probe with typical D-π-A structure was designed and showed high sensitivity to polarity and viscosity changes. DPAR could selectively detect human serum albumin (HSA) with turn-on orange emission in aqueous PBS buffer (pH 7.4), which showed advantages such as rapid response (4 min), high sensitivity (LOD 0.98 μg/mL). Therefore, it was successfully used for achieving HSA levels in urine samples and HSA imaging in HeLa cells. DPAR also exhibited the capability to recognize the cancer cells over the normal cells by lower polarity guided lipid droplets (LDs) imaging (in green emission channel). The detection mechanism for HSA and cancer diagnosis was convinced that DPAR encountered the lower-polarity and higher-viscosity microenvironment, resulting in the confinement of the TICT process and intramolecular rotation. These facts showed that DPAR had good application prospects in environment-related biomedical research and clinical diagnosis.

An infinite chain, {[Ni(tn)2]3[Fe(CN)4(μ-CN)2]2}n, a new catalyst for electrochemical-driven water splitting and photochemical-driven hydrogen evolution from water under blue light

A new catalyst for both water reduction and oxidation, based on an infinite chain, {[Ni(tn)2]3 [Fe(CN)4 (μ-CN)2]2}n, is formed by the reaction of NiCl2, 1,3-propanediamine (tn) and K3 [Fe(CN)6]. {[Ni(tn)2]3 [Fe(CN)4 (μ-CN)2]2}n can electro-catalyze hydrogen evolution from a neutral aqueous buffer (pH 7.0) with a turnover frequency (TOF) of 1561 mol of hydrogen per mole of catalyst per hour (H2/mol catalyst/h) at an overpotential (OP) of 837 mV {[Ni(tn)2]3 [Fe(CN)4 (μ-CN)2]2}n also can electro-catalyze O2 production from water with a TOF of ∼45 mol O2 (mol cat)−1s−1 at an OP of 591 mV. Under blue light (λ = 469 nm), together with CdS nanorods (CdS NRs) as a photosensitizer, and ascorbic acid (H2A) as a sacrificial electron donor, {[Ni(tn)2]3 [Fe(CN)4 (μ-CN)2]2}n can photo-catalyze hydrogen generation from an aqueous buffer (pH 4.0) with a turnover number (TON) of 11,450 mol H2 per mole of catalyst (mol of H2 (mol of cat)−1) during 10 h irradiation. The average of apparent quantum yield (AQY) is as high as 40.96% during 10 h irradiation. Studies indicate that {[Ni(tn)2]3 [Fe(CN)4 (μ-CN)2]2}n exists in two forms: a cyano-bridged chain ({[Ni(tn)2]3 [Fe(CN)4 (μ-CN)2]2}n) in solid, and a salt ([Ni(tn)2]3 [Fe(CN)6]2) in aqueous media; Catalytic reaction occurs on the nickel center of [Ni(tn)2]2+, and the introduction of [Fe(CN)6]3- can improve the catalytic efficiency of [Ni(tn)2]2+ for H2 or O2 generation. We hope these findings can afford a new method for the design of catalysts for both water reduction and oxidation.

Solvent and temperature effects on the solubility of some new adamantane/memantine sulfonamide derivatives

The dissolution processes of five new adamantane/memantine sulfonamide derivatives with different substituents in the phenyl ring were examined in the pharmaceutically relevant solvents: aqueous buffer pH 7.4, 1-octanol and n-hexane by the shake-flask method in the temperature range of 293.15–313.15 K. Compounds revealed low equilibrium solubility in water and n-hexane, and essentially better solubility in 1-octanol. Experimental solubility was correlated with the melting properties (measure of crystal lattice energy) and hypothetic distribution coefficient (measure of solvation). Neuroprotective potential of the investigated derivatives was accessed with the help of ΔlogD parameter. The solubility evaluation using the Hansen solubility parameters showed the consistency with the experimental data and revealed the trends in the structure modification within the studied class in order to achieve good potential adsorption. The activity coefficients at infinite dilution, ideal solubility of the compounds in the solvents and molar excess thermodynamic functions have been calculated. Ideal solubility was recorded much higher than the experimental one at each temperature. It has been estimated that in most cases the entropy term of the excess Gibbs energy is responsible for the deviation from ideality in the investigated drug-solvent systems. The solubility data and thermodynamic properties obtained would be useful in the process of purification, recrystallization and dosage form design of adamantane/memantine sulfonamide derivatives in pharmaceutical industry.

Electrochemical Oxidation of 2,3-Dihydroxypyridine in the Presence of Benzenesulfinic Acid: A Green Method for the Synthesis of a Novel Heterocycle Dye

A green method for the one-pot electrochemical synthesis of a novel highly oxygenated heterocycle dye, 3,8-dihydroxy-4,9-bis(phenylsulfonyl)dipyrano[2,3-b:2′,3′-e]pyrazine-2,7-dione (PSO), is reported. The synthesis of this compound was performed by anodic oxidation of 2,3-dihydroxypyridine (DHP) in the presence of benzenesulfinic acid (BSA) in aqueous acetate buffer solution, using a carbon electrode. The cyclic voltammogram of DHP shows an irreversible anodic peak corresponding to two-electron oxidation of DHP to pyridine-2,3-dione (PDO), while, the CV of the mixture of DHP and BSA exhibits a new anodic peak at more positive potentials which is a confirmation of the reaction between electrogenerated PDO and BSA. Based on the electrochemical evidence, a novel mechanism for the synthesis of PSO is proposed. The synthesis of PSO was conducted in aqueous acetate buffer (pH = 4.0, c = 0.2 M), by controlled potential method (at 0.90 V vs Ag/AgCl electrode, yield 69%) as well as by constant current method (at J = 1.12 mA cm−2, yield 63%) in a divided cell, under green conditions. Furthermore, the pH dependent anodic behavior of 2,3-dihydroxypyridine was investigated. Our data shows that the E pA–pH diagram indicates the presence of five predominant regions. The predominant species across all regions were identified.

Synthesis and Guest-Binding Properties of pH/Reduction Dual-Responsive Cyclophane Dimer.

A water-soluble cyclophane dimer having two disulfide groups as a reduction-responsive cleavable bond as well as several acidic and basic functional groups as a pH-responsive ionizable group 1 was successfully synthesized. It was found that 1 showed pH-dependent guest-binding behavior. That is, 1 strongly bound an anionic guest, 6-p-toluidinonaphthalene-2-sulfonate (TNS) with binding constant (K/M−1) for 1:1 host-guest complexes of 9.6 × 104 M−1 at pH 3.8, which was larger than those at pH 7.4 and 10.7 (6.0 × 104 and 2.4 × 104 M−1, respectively), indicating a favorable electrostatic interaction between anionic guest and net cationic 1. What is more, release of the entrapped guest molecules by 1 was easily controlled by pH stimulus. Large favorable enthalpies (ΔH) for formation of host-guest complexes were obtained under the pH conditions employed, suggesting that electrostatic interaction between anionic TNS and 1 was the most important driving force for host-guest complexation. Such contributions of ΔH for formation of host-guest complexes decreased along with increased pH values from acidic to basic solutions. Upon addition of dithiothreitol (DTT) as a reducing reagent to an aqueous PBS buffer (pH 7.4) containing 1 and TNS, the fluorescence intensity originating from the bound guest molecules decreased gradually. A treatment of 1 with DTT gave 2, having less guest-binding affinity by the cleavage of disulfide bonds of 1. Consequently, almost all entrapped guest molecules by 1 were released from the host. Moreover, such reduction-responsive cleavage of 1 and release of bound guest molecules was performed more rapidly in aqueous buffer at pH 10.7.