Stern-Volmer Plots Research Articles

Exploring the dual sensing properties of an anthraldehyde based Schiff base for the successive determination of picric acid using AIEEF and copper using colorimetric methods

A novel and lucid Schiff base, designated as AHN, exhibiting multiple analytical responses comprising AIEE (Aggregation Induced Emission Enhancement) and colorimetric activity towards distinct analytes has been designed and synthesized. The probe, AHN, effectively leverages AIEE activity to selectively detect picric acid (PA) on behalf of fluorescence switch-off response amidst other nitro chemicals. Additionally, it offers remarkable colorimetric selectivity to bivalent copper ions (Cu2+) via a distinct colour change from colourless to yellow. The multi-response characteristics of AHN propel its practicality for naked-eye detection of these analytes. The addition of PA to the aggregate of AHN in DMSO with a 90% water fraction induces a quenching in the fluorescence intensity of the AHN aggregate and the limit of detection of PA was found to be 24.5 × 10−7 M with a quenching constant value of 6.21 × 107 M−1. Stern Volmer plots and lifetime measurements, clearly indicate that both static and dynamic processes were involved in the quenching mechanism. It is also evident that ground-state complexation between electron-rich fluorescent aggregates of AHN and electron-deficient PA takes place through π-π interactions and intramolecular hydrogen bonding interactions. Further, AHN in DMSO exhibits a selective colorimetric response to bivalent copper among various metal ions with a detection limit of 3.16 µM which is substantially below the permitted limit recommended by WHO in drinking water. The mechanism of colorimetric response is the complexation of AHN with Cu2+ in the 2:1 stoichiometry, as confirmed by Job’s plot method. Consequently, the versatility of probe AHN in detecting PA and Cu2+ through distinct mechanisms fosters its significance in the field of sensing and opens promising avenues for practical sensing applications.

Utility of the food colorant erythrosine B as an effective green probe for quantitation of the anticancer sunitinib. Application to pharmaceutical formulations and human plasma.

Sunitinib is a tyrosine kinase inhibitor used for the treatment of renal cell carcinoma and gastrointestinal stromal tumors. In this study, two spectroscopic methods, spectrofluorometric and spectrophotometric, were utilized to quantify sunitinib in different matrices. In method I, the native fluorescence of erythrosine B was quenched by forming ion-pair complex with increasing quantities of sunitinib. This approach was utilized for measuring sunitinib in its dosage forms and spiked plasma. After excitation at 528 nm, the quenching of fluorescence is linearly related to the concentration across the range of 0.05-0.5μg mL-1 at 550 nm in Britton-Robinson buffer (pH4.0), with a correlation value of 0.9999 and a high level of sensitivity with detection limit down to 10ng mL-1 . Method II relies on spectrophotometric measurements of the produced complex at 550 nm across a range of 0.5-10.0μg mL-1 , with good correlation value of 0.9999. This method has a detection limit down to 0.16 μg mL-1 . The proposed methodologies were validated according to International Conference on Harmonization (ICH) guidelines with satisfactory results. The stoichiometry of the reaction was determined through the application of Job's method, while the mechanism of quenching was investigated by employing the Stern-Volmer plot. The designated methods were used to estimate sunitinib in its capsules and in spiked human plasma. Additionally, the statistical analysis of the data revealed no substantial differences when compared to previous reported spectroscopic method. Green assessment tools provide further details about the eco-friendly nature of the methods.

Ecofriendly Approach for the Determination of Selected Aldehydes by Fluorescence Quenching of L-Tryptophan.

It is a fluorescence-based study to examine the interaction between L-tryptophan and a selection of aldehydes, namely furfural (furan-2-carbaldehyde), 3-hydroxybenzaldehyde, salicylaldehyde (2-hydroxybenzaldehyde), 3-nitrobenzaldehyde, and 4-bromobenzaldehyde. The investigation took place in an aqueous environment, revealing that all five aldehydes induced quenching of the fluorescence intensity of L-tryptophan. By employing the Stern-Volmer equation to describe the quenching process, we constructed Stern-Volmer plots and derived Stern-Volmer constants. These constants (KSV) ranged from 2.87 × 104mol L- 1 to 5.75 × 104mol L- 1. Notably, the values of the Stern-Volmer constants varied among the different aldehydes, with the following order: 3-hydroxybenzaldehyde(3-HBA) > 4-bromobenzaldehyde (4-BBA) > 3-nitrobenzaldehyde > furan-2-carbaldehyde > salicylaldehyde. Consequently, our findings highlighted 3-hydroxybenzaldehyde as the most potent quencher, while 2-hydroxybenzaldehyde displayed the least sensitivity to quenching. Additionally, we determined the detection and quantification limits for the investigated aldehydes, resulting in ranges of 3.87 × 10- 12 to 8.25 × 10- 6 and 1.29 × 10- 11 to 2.75 × 10- 5, respectively. This research paves the way for the development of novel fluorescence probe-based sensors and offers valuable techniques for analyzing aldehydes within environmental and biological samples.

Profiling in-vitro release of verteporfin from VISUDYNE® liposomal formulation and investigating verteporfin binding to human serum albumin

VISUDYNE® is a liposomal formulation of verteporfin, used in the photodynamic therapy of age-related macular degeneration via intravenous administration. In this study, we developed a new in vitro method to quantify verteporfin release from VISUDYNE® under conditions that replicate in vivo conditions using human serum albumin (HSA). Verteporfin release from the liposomes was quantified using capillary electrophoresis (CE) with optical detection. Verteporfin binding to HSA was quantified by measuring HSA fluorescence that is quenched by drugs binding to specific HSA binding sites. The binding constant of verteporfin to HSA was calculated using the Stern Volmer plot and found to be 1.966 × 107 M−1 at 37 °C. Verteporfin binding to HSA involves one albumin binding site and the binding molar ratio between verteporfin and HSA is approximately 1:1. A rapid partitioning of verteporfin from VISUDYNE® onto HSA takes place within 10 min and involves the release of more than 90% of the verteporfin at physiological temperatures. This study verifies this approach of using CE to rapidly separate liposome and HSA-bound drug, thus minimizing drug release artifacts created with other methods.

Hybridization of quinacridone and synthetic hectorite and the photoluminescence quenching by metal ions

Quinacridone, a red pigment, was hybridized with a synthetic hectorite by a mechanochemical reaction. Though quinacridone was not photoluminescent, the products were photoluminescent and the photoluminescence quantum yield was 0.89 when the dye loading was 0.00009 g dye/g clay. The product was dispersed in water to give stable suspensions with varied viscosities depending on the concentration of the hybrids. The photoluminescence of the aqueous suspension was quenched by Cr3+, Co2+, Cu2+, Fe2+, Ni2+ and Pb2+ as described by linear Stern-Volmer plots at the concentration of 1–600 ppm. Among the tested metal ions, Cr3+ gave the most efficient quenching (the KSV value of 0.0125). The quenching of the photoluminescence of the quinacridone-synthetic hectorite hybrid was applicable to detect Cu2+ and Cr3+ at ppb level.

Direct Evidence for Excited Ligand Field State-based Oxidative Photoredox Chemistry of a Cobalt(III) Polypyridyl Photosensitizer.

Increasing interest in sustainable chemistry coupled with the quest to explore new reactivity has spurred research on first-row transition metal complexes for potential applications in a variety of settings. One of the more active areas of research is photoredox catalysis, where the synthetically tunable nature of their electronic structures provides a rich palette of options for tailoring their reactivity to a desired chemical transformation. Understanding the mechanism of excited-state reactivity is critical for the informed development of next-generation catalysts, which in turn requires information concerning the propensity of their electronic excited states to engage in the desired electron or energy transfer processes. Herein we provide direct evidence of the highly oxidizing nature of the lowest-energy ligand-field (LF) excited state of a first-row d6-low-spin Co(III) photosensitizer [Co(4,4'-Br2bpy)3]3+ (where 4,4'-Br2bpy is 4,4'-dibromo-2,2'-bipyridine). The redox potential associated with the LF excited state of the Co(III) complex was bracketed by performing bimolecular quenching studies by using a series of simple organic electron donors. Time-resolved absorption spectroscopy confirmed a dynamic quenching process attributed to reductive quenching of the lowest-energy ligand-field excited state of the Co(III) chromophore. Analysis of the Stern-Volmer plots for each chromophore-quencher pair revealed a limiting value of Ered* ∼ 1.25 V vs Fc/Fc+ for the metal-centered excited state, which is significantly stronger than that of more commonly employed transition metal-based photoredox agents such as [Ru(bpy)3]2+ (Ered* = 0.32 V vs Fc/Fc+) and [Ir(ppy)2(bpy)]+ (Ered* = 0.27 V vs Fc/Fc+). These results suggest that this class of chromophores could find utility in applications requiring the activation of oxidatively resistant organic substrates for photoredox catalysis.

Fluorescence Quenching Study Of 5-Sulphosalicylic Acid Doped Polyaniline-Γ-Fe2o3 Composite For High Energy Material Sensing (Fe-Spani)

Background: Polymeric nanocomposites containing metal oxides are of interest to researchers because these materials show hybrid properties that are generated from both components. Due to the control and design of structural properties at the nanoscale scale, they have specialized properties for a range of applications. Researchers have tried to improve the desirable characteristics and thus increase their application by reinforcing them with nanoscale materials to provide them with better properties than conventional micro composites. Objective: In our present work we have used 5-sulphosalicylic acid doped polyaniline-γ-Fe2O3 composite (Fe-SPAni) to detect explosives using fluorescence quenching method. Along with ease of synthesis and low cost, Fe-SPAni possesses a variable oxidation state which is important for trace detection of the analytes. Methods: We used the fluorophore Fe-SPAni to detect High Energy Materials (HEMs) using the fluorescence quenching approach, which is still difficult to achieve. UV-Vis spectroscopy, Fourier-transform infrared (FTIR) spectroscopy, and scanning electron microscopy (SEM/EDAX) were used to determine the molecular structure of the Fe-SPAni composite produced by oxidative chemical polymerization; CV studies were performed to investigate the electrochemical properties. HEMs such as dinitrobenzene (DNB), pentaerythritol tetranitrate (PETN), trinitrotoulene (TNT), 1,3,5-trinitroperhydro-1,3,5-triazine (RDX), and hexanitrohexaazaisowurtzitane(CL-20) were used as quenchers.The fluorescence quenching process was determined using the Stern-Volmer graph. Results: The Stern-Volmer(S-V) plot and limit of detection (LOD) data showed that PETN had the greatest results for the quenching constant value (Ksv) (Ksv =1.33 x 106 M-1 and 1.4x 10-6M, respectively). Conclusion: The research points to Fe-SPAni as a promising contender for sensing explosives.

Highly Efficient Fluorescent Detection of Vitamin B12 Based on the Inner Filter Effect of Dithiol-Functionalized Silver Nanoparticles.

We report a fluorescent assay for the determination of vitamin B12 (VB12) based on the inner filter effect (IFE) of 1,3-propanedithiol-functionalized silver nanoparticles (PDT-AgNPs). PDT was simply functionalized on the surface of AgNPs through Ag-thiol interaction, which leads to significantly enhanced fluorescence, with excitation and emission at 360 and 410 nm, respectively, via their thiol-mediated aggregation. Since target VB12 has strong absorption centered at 360 nm, which is almost completely overlapping with the excitation spectra of PDT-AgNPs, the VB12 induced strong quenching of the fluorescence of PDT-AgNPs via IFE. The IFE-based mechanism for the fluorescence quenching of PDT-AgNPs in the presence of VB12 was confirmed by the analyses of Stern-Volmer plots at different temperatures and fluorescence decay curves. The fluorescence-quenching efficiency of PDT-AgNPs was linearly proportional to the concentration of VB12 in a wide range of 1 to 50 μM, with a lower detection limit of 0.5 μM, while preserving excellent selectivity toward target VB12 among possible interfering molecules. Furthermore, the PDT-AgNPs-mediated assay succeeded in quantitatively detecting VB12 in drug tablets, indicating that PDT-AgNPs can serve as an IFE-based fluorescent probe in pharmaceutical preparations by taking advantages of its ease of use, rapidity, and affordability.

Evidence of enhanced inhibitory potential of Ferulic Acid Ethyl Ester towards modulating fibrillation and disassembling insulin fibrils

The conversion of native conformation of protein into proteotoxic insoluble entities is not only confined to the protein misfolding diseases(PMDs) but also associated with pharmaceutically important proteins like insulin during the process of manufacturing/administration. Also all the neurodegenerative diseases are marked with increased level of oxidative stress. Thus finding anti-aggregation agents having anti-oxidant potential is utmost important to combat these complications. Herein, we performed biophysical, computational and imaging techniques to decipher the anti-aggregation propensity of a naturally occurring hydroxycinnamate- Ferulic acid ethyl ester(FAEE) that can cross blood brain barrier. Rayleigh light scattering(RLS) measurements, dye binding assays, Circular dichroism(CD) and Dynamic light scattering(DLS) measurements confirmed the efficient inhibition of insulin fibrillation by FAEE. The Stern-Volmer plot analysis reflected both the static and dynamic quenching mechanism of insulin by FAEE. Molecular docking and simulations results showed that FAEE bind and stabilizes the monomeric state of insulin with binding energy −6.3 kcal/mol. Moreover, the inhibiting effect of FAEE on the elongation phase of onward fibrillation in addition with triggering its fibril disassembling capability was also investigated. Both the stabilization of the protein structure and the restriction of monomers recruitment necessary for the fibrillar growth acts synergistically in providing FAEE its anti-aggregation effect. Appearance of significantly lesser and shortened fibrils in FAEE presence was depicted by transmission electron microscopy. Overall, our finding establishes for the first time the anti-amyloidogenic role of lipophilic FAEE which can cross blood brain barrier. These findings may inspire the development of FAEE based formulations that could possibly treat amyloid-related diseases.

The structure of model and peptide disulfides markedly affects their reactivity and products formed with singlet oxygen

Disulfide bonds are critical structural elements in proteins and stabilize folded structures. Modification of these linkages is associated with a loss of structure and function. Previous studies have reported large variations in the rate of disulfide oxidation by hypohalous acids, due to stabilization of reaction intermediates. In this study we hypothesized that considerable variation (and hence selective oxidation) would occur with singlet oxygen (1O2), a key intermediate in photo-oxidation reactions. The kinetics of disulfide-mediated 1O2 removal were monitored using the time-resolved 1270 nm phosphorescence of 1O2. Stern-Volmer plots of these data showed a large variation (∼103) in the quenching rate constants kq (from 2 × 107 for α-lipoic acid to 3.6 × 104 M−1s−1 for cystamine). The time course of disulfide loss and product formation (determined by LC-MS) support a role for 1O2, with mono- and di-oxygenated products detected. Elevated levels of these latter species were generated in D2O- compared to H2O buffers, which is consistent with solvent effects on the 1O2 lifetime. These data are interpreted in terms of the intermediacy of a zwitterion [–S+(OO−)–S–], which either isomerizes to a thiosulfonate [–S(O)2–S–] or reacts with another parent molecule to give two thiosulfinates [–S(O)–S-]. The variation in quenching rates and product formation are ascribed to zwitterion stabilization by neighboring, or remote, lone pairs of electrons. These data suggest that some disulfides, including some present within or attached to proteins (e.g., α-lipoic acid), may be selectively modified, and undergo subsequent cleavage, with adverse effects on protein structure and function.

A facile and highly selective fluorimetric chemosensor 1,2,4-Aminonaphthol sulfonic acid for detection of copper ions in aqueous medium

1,2,4-Aminonaphthol sulfonic acid (H) shows strong fluorimetric properties which can be utilized for recognition behavior of cation identification. Sensor (H) has selective complexing ability towards Cu2+ ions in presence of other metal ions in aqueous medium. Study of Stern-Volmer plot confirmed static as well as dynamic quenching mechanisms. The Job's plot exhibited stoichiometric ratio between metal ions and receptor as 1:1. Benesi Hildebrand equation was used to calculate association constant Ka (1.43 × 103 M-1) for Cu2+ ions. Fluorescence study revealed that the sensor (H) is capable to detect Cu2+ ions efficiently in aqueous medium without the bar of maintaining pH of the solution.

Environmentally friendly protocol for the determination of sitagliptin phosphate in pharmaceutical preparations and biological fluids using l-tyrosine as a fluorescence probe.

A responsive spectrofluorometric method was developed for the determination of sitagliptin phosphate using l-tyrosine as a fluorescence probe. The fluorescence intensity of l-tyrosine was quenched with sitagliptin phosphate. The fluorescence intensity was recorded at 307 nm using a 272 nm excitation wavelength. The calibration plot between fluorescence intensity and the concentration of drug was linear in the range of 0.1 to 2.0mM with a good correlation value of 0.997. The limit of detection and quantification were established to be 3.7×10-4 and 1.23 ×10-3 mM, respectively. Commonly used excipients did not interfere with sitagliptin phosphate measurement. The proposed method was used to measure the sitagliptin phosphate in its standard type, dosage form, and biological samples. The percent recovery ranged from 97.41-103.36%. The static quenching was shown to be responsible for quenching as indicated by the Stern-Volmer plot. The method was validated using ICH guidelines and profitably applied for the content uniformity test, resulting in a high percent recovery and small relative standard deviation. The proposed approach is effortless, susceptible, selective, economic, and provides a high precision and accuracy, and can be used to determine sitagliptin phosphate in the pharmaceutical industry.

Optical Dual Gas Sensor for Simultaneous Detection of Nitric Oxide and Oxygen

This work presents a new optical dual sensor based on PtTFPP-containing electrospun fibers and CsPbBr3 perovskite quantum dots (PQDs) for simultaneous detection of oxygen (O2) and nitric oxide (NO) gases, wherein PtTFPP-containing electrospun fibers for O2 sensing was based on electrospinning process fabricated by platinum(II) meso-tetrakis (pentafluorophenyl) porphyrin (PtTFPP) complex immobilized in cellulose acetate (CA) matrix. CsPbBr3 PQDs were used as NO-sensitive material and coated on the surface of PtTFPP-containing electrospun fibers. Both materials were excited by a UV LED with a central wavelength of 380 nm, and the fluorescence intensities of sensing materials were recorded and analyzed with a spectrometer. The experiment results show that the optical NO and O2 sensors have linear Stern–Volmer plots, and the sensitivities are around 2.7 and 10.7, respectively. The response and recovery times of the optical NO sensor are 71 and 109 s, respectively. For optical O2, response and recovery times are 60 and 65 s, respectively. The optical dual sensor with a new method based on fluorescent dye containing electrospun fibers and coated with CsPbBr3 PQDs has been successfully developed to detect NO and O2 gases simultaneously. The optical dual gas sensor provides great potential for practical applications with low cost and ease of fabrication.

Biophysical studies of the binding of histone deacetylase inhibitor (Trichostatin-A) with bovine serum albumin

Trichostatin A (TSA), a potential radiomitigator in pre-clinical models, inhibits the class I and II mammalian histone deacetylase (HDAC) enzyme family preferentially. In the current study, the ADME assessment of TSA was explored in terms of its binding affinity for serum protein via spectroscopic and molecular docking techniques. Fluorescence spectroscopy was used to examine changes in the protein microenvironment, and affinity was quantified in terms of binding constant and stoichiometry. Post binding conformational changes were observed using circular dichroism (CD) and UV-Visible spectroscopy. Specific binding was visualized using molecular docking to support experimental studies. UV-vis spectra demonstrated a blue shift in the interaction of TSA to BSA. The calculated binding constants ranged from 3.10 to 0.78 x 10 5(M−1) and quenching constants from 2.75 to 2.15 x 104 (l mol-1), indicating TSA has a strong binding affinity for BSA. Based on the FRET theory, the distance between BSA (donor) and TSA (acceptor) was calculated to be 2.83 nm. The Stern-Volmer plot revealed (Ksv) static quenching. Thermodynamic parameters were calculated, and a negative ΔG value showed that the interaction is spontaneous. The CD spectra analysis further revealed a change in the protein’s secondary structure, indicating TSA-BSA interaction. The molecular docking studies also indicated strong binding affinity of TSA with BSA. The results indicate that good bio-availability of TSA is possible because of the spontaneous and strong binding affinity with BSA. Communicated by Ramaswamy H. Sarma

Acriflavine: an efficient green fluorescent probe for sensitive analysis of aceclofenac in pharmaceutical formulations

Acriflavine is a multipurpose drug that shows antibacterial, antiviral, antimalarial, and antifungal activities. The remarkable native fluorescence of acriflavine is exploited in analytical chemistry field as an efficient probe for analysis of pharmaceutical and biological compounds. The fluorescent probe action of acriflavine is based on the remarkable fluorescence turning-off via formation of ion-pair complexes with acidic drugs at a specific pH. Herein, the acidic drug aceclofenac is analysed for the first time using acriflavine as a fluorescent probe. Aceclofenac can form an ion-pair complex with acriflavine at pH 8.5, and hence it partially turns off the fluorescence intensity of acriflavine over a concentration range of 1–20 µg/mL. The fluorescence quenching was monitored at 502 nm following an excitation at 265 or 451 nm. The reaction stoichiometry between acriflavine and aceclofenac was found to be 1:1 using limiting logarithmic method. The type of quenching was confirmed to be static using Stern–Volmer plot. The method showed low values of quantitation limit (0.89 µg/mL) and detection limit (0.29 µg/mL). Moreover, the method was linear (r = 0.9999), accurate, precise (RSD < 1.7%), robust, and specific. The proposed method was successfully employed to analyse aceclofenac in its dosage forms with high %recovery (98–101%). Additionally, GAPI and AGREE approaches were used to guarantee the suggested techniques' greenness, and the findings showed an excellent level of greenness.

Cellulose/amylose derivatives bearing bulky substituents as reversible fluorescent sensors for detection of Fe3+

Two novel cellulose and amylose derivatives bearing bulky tris(2-benzothienylformate) pendants (Cel-3 and Amy-3) were expeditiously prepared by one-step esterification. The fluorescent sensing performance of six polysaccharide derivatives, including Cel-3/Amy-3, and other four previously prepared benzothienyl- or benzofuranyl-phenylcarbamates of cellulose and amylose (Cel-1/Amy-1, Cel-2/Amy-2), were carefully evaluated using eight metal ions, including Co2+, K+, Na+, Li+, Hg2+, Ni2+, Ca2+ and Fe3+. All six derivatives exhibited excellent fluorescence quenching property to Fe3+ ions with high sensitivity and selectivity. Especially, the limit of detection of Amy-2 with benzofuranylphenylcarbamates for Fe3+ was 3.0 μM, much lower than the maximum contaminant level for Fe3+ in the drinking water. Additionally, the six bulky derivatives displayed the interesting fluorescence “turn-off” and “turn-on” observation, indicating a desirable reversibility for Fe3+ detection. The high anti-interference ability was also observed for detection of Fe3+ on the benzothienyl/benzofuranyl derivatives of cellulose and amylose in the combined system containing Co2+, K+, Na+, Li+, Hg2+, Ni2+ and Ca2+. It suggested that the obtained polysaccharide derivatives with bulky chromophores possessed good potentials for detection of Fe3+ as high-efficient fluorescent sensors in diverse applications. The sensing mechanism for detection of Fe3+ was further proposed based on the Stern-Volmer plots and fluorescence titration analysis.

Interaction studies of cannabidiol with human serum albumin by surface plasmon resonance, spectroscopy, and molecular docking

The binding interaction of cannabidiol (CBD) and human serum albumin (HSA) under physiological blood pH conditions (pH 7.4) was conducted by surface plasmon resonance (SPR), fluorescence spectroscopy, UV-Visible spectrophotometry, and molecular docking. The responses from SPR measurement increased with the increase in CBD concentration until equilibrium was reached at the equilibrium dissociation constant (KD) of 9.8 × 10−4 M. The results from fluorescence and UV-Visible spectroscopy showed that CBD bound to HSA at one site in a spontaneous manner to form protein-CBD complexes. The quenching process involved both static and dynamic mechanisms while the static mechanism contributed predominantly to the binding between CBD and albumin. The binding constants estimated from the fluorescence studies were from 0.16 × 103 to 8.10 × 103 M−1, which were calculated at different temperature conditions using Stern-Volmer plots. The thermodynamic parameters demonstrated that the binding interaction was a spontaneous reaction as Gibbs free energy had negative values (ΔG = −12.57 to −23.20 kJ.mol−1). Positive ΔH and ΔS values (ΔH = 2.46 × 105 J.mol−1 and ΔS = 869.81 J.mol−1K−1) indicated that the hydrophobic force was the major binding interaction. Finally, confirmation of the type and extent of interaction was provided using UV-spectroscopy and molecular docking studies. The outcomes of this study are expected to serve as a platform to conduct future studies on binding interactions and toxicological research of CBD. Communicated by Ramaswamy H. Sarma

Exploring the binding behavior mechanism of vitamin B12 to α-Casein and β-Casein: multi-spectroscopy and molecular dynamic approaches

The aim of this study was to investigate the behavior interaction of α-Casein-B12 and β-Casein-B12 complexes as binary systems through the methods of multiple spectroscopic, zeta potential, calorimetric, and molecular dynamics (MD) simulation. Fluorescence spectroscopy denoted the role ofB12as a quencher in both cases of α-Casein and β-Casein fluorescence intensities, which also verifies the existence of interactions. The quenching constants of α-Casein-B12 and β-Casein-B12 complexes at 298 K in the first set of binding sites were 2.89 × 104 and 4.41 × 104 M−1, while the constants of second set of binding sites were 8.56 × 104 and 1.58 × 105 M−1, respectively. The data of synchronized fluorescence spectroscopy at Δλ = 60 nm were indicative of the closer location of β-Casein-B12 complex to the Tyr residues. Additionally, the binding distance between B12 and the Trp residues of α-Casein and β-Casein were obtained in accordance to the Förster’s theory of nonradioactive energy transfer to be 1.95 nm and 1.85 nm, respectively. Relatively, the RLS results demonstrated the production of larger particles in both systems, while the outcomes of zeta potential confirmed the formation of α-Casein-B12 and β-Casein-B12 complexes and approved the existence of electrostatic interactions. We also evaluated the thermodynamic parameters by considering the fluorescence data at three varying temperatures. According to the nonlinear Stern-Volmer plots of α-Casein and β-Casein in the presence of B12 in binary systems, the two sets of binding sites indicated the detection of two types of interaction behaviors. Time-resolved fluorescence results revealed that the fluorescence quenching of complexes are static mechanism. Furthermore, the outcomes of circular dichroism (CD) represented the occurrence of conformational changes in α-Casein and β-Casein upon their binding to B12 as the binary system. The experimental results that were obtained throughout the binding of α-Casein-B12 and β-Casein-B12 complexes were confirmed by molecular modeling. Communicated by Ramaswamy H. Sarma

Revelation of anti-aggregating and disaggregating characteristic of amodiaquine via inhibition of human lysozyme fibrillation: Therapeutic alternative for treating amyloidopathies

Amyloidopathies are associated with a biochemical change in concerned organ or bio-environment known as inflammation (promoted in case of fibrils accumulation due to their stability) that triggered us to explore anti-aggregating and disaggregating property of an anti-inflammatory drug amodiaquine (AD) against human lysozyme (HL) fibrillation in this study simultaneously evaluating the mechanism of interaction between AD and HL using biophysical and microscopic techniques. The linear modified Stern-Volmer plot obtained from fluorescence quenching of HL by AD at three temperatures and Ksv dependence on temperature with kq > 2.0 × 1010 supports static quenching mechanism with strong binding (Kb = × 106) and negative ΔG value. The change in Trp and Tyr environment due to AD binding were confirmed via synchronous fluorescence. The studies were further extended to analyse aggregation inhibition and disaggregation of HL by AD. RLS, ThT, CD and DLS results suggested aggregation inhibition and disaggregation which was further confirmed via TEM. Overall, our study suggests that AD is an effective drug for fibrillation inhibition and disaggregation. The possible mechanism behind this behaviour is the stabilization of HL in native state by AD via non-covalent interactions and thus AD will not only reduce fibril burden but will also calm severity of the disease (such as regaining cognitive function due to reduced inflammation). This can be an alternative promising approach that can be exploited for treating amyloidopathies such as Alzheimer’s, Parkinson’s disease and systemic amyloidosis.

Ultrasound-Assisted Synthesis of Chalcone: A Highly Sensitive and Selective Fluorescent Chemosensor for the Detection of Fe3+ in Aqueous Media.

The chalcone compound DHPO was synthesized through a chemical reaction between 1-(2-hydroxyphenyl)-ethanone and 3,4-dimethoxy benzaldehyde under ultrasound irradiation. The interaction between the DHPO compound and several metal ions was studied using fluorescence behavior, revealing that the chalcone function as a "turn on and turn off" switch fluorescent sensor, for selectively and sensitively detecting Fe3+ ions. The process of fluorescence quenching and complexation of DHPO with Fe3+ ion was further studied using methods such as Benesi-Hildebrand, Stern-Volmer plot, and job plot.