A rapid, simple and ultrasensitive spectrofluorimetric method for the direct detection of metformin in real samples based on a nanoquenching approach.

Metformin (MET), as an oral antidiabetic and antihyperglycemic agent, is widely used to treat type II diabetes mellitus. Because of its increasing consumption, developing a fast, simple, and selective method to determine its concentration in biological samples (serum and urine) and pharmaceutical formulations (tablets) is of great interest. In this study, we used a FRET-based fluorescent nanosensor (Tb-phen-AgNPs system) for sensitive detection of MET in tablet and serum samples. This method is based on the enhancing effect of MET on the emission intensity of the Tb-phen complex, which is quenched by AgNPs via energy transfer process (turn off-on mode). A good linear relationship between the MET concentration and enhanced emission intensity of the Tb-phen-AgNPs system was observed in the range of (0.75-3.7) × 10-6 M under optimum conditions. Limit of detection and limit of quantitation were calculated to be 0.43 × 10-6 M and 1.31 × 10-6 M, respectively. This method was successfully used to determine MET concentrations in pharmaceutical dosage form and in spiked serum sample. The obtained recoveries from pharmaceutical formulation and serum sample were in the range 86.75-98.97% and 85.10-100.96%, respectively. Collectively, our results indicated that the method described here is simple, sensitive, cost effective, and free from interference. Therefore, it can be used as an effective and routine method for the direct and rapid determination of MET levels in biological samples such as serum.

Exploring the interactions of a Tb(III)-quercetin complex with serum albumins (HSA and BSA): spectroscopic and molecular docking studies.

Serum albumins (human serum albumin (HSA) and bovine serum albumin (BSA), two main circulatory proteins), are globular and monomeric macromolecules in plasma that transport many drugs and compounds. In the present study, we investigated the interactions of the Tb(III)-quercetin (Tb-QUE) complex with HSA and BSA using common spectroscopic techniques and a molecular docking study. Fluorescence data revealed that the inherent fluorescence emission of HSA and BSA was markedly quenched by the Tb-QUE complex through a static quenching mechanism, confirming stable complex formation (a ground-state association) between albumins and Tb-QUE. Binding and thermodynamic parameters were obtained from the fluorescence spectra and the related equations at different temperatures under biological conditions. The binding constants (Kb ) were calculated to be 0.8547 × 103 M-1 for HSA and 0.1363 × 103 M-1 for BSA at 298 K. Also, the number of binding sites (n) of the HSA/BSA-Tb-QUE systems was obtained to be approximately 1. Thermodynamic data calculations along with molecular docking results indicated that electrostatic interactions have a main role in the binding process of the Tb-QUE complex with HSA/BSA. Furthermore, molecular docking outputs revealed that the Tb-QUE complex has high affinity to bind to subdomain IIA of HSA and BSA. Binding distances (r) between HSA-Tb-QUE and BSA-Tb-QUE systems were also calculated using the Forster (fluorescence resonance energy transfer) method. It is expected that this study will provide a pathway for designing new compounds with multiple beneficial effects on human health from the phenolic compounds family such as the Tb-QUE complex.

Probing the interaction between 7-geranyloxycoumarin and bovine serum albumin: Spectroscopic analyzing and molecular docking study.

7-Geranyloxycoumarin (auraptene; AUR), as a potent phytochemical, is the naturally abundant prenyloxycoumarin found in many genera of the Rutaceae family. As the interaction with serum albumins may play a crucial role in identifying their pharmacological properties, we investigated AUR binding profile with bovine serum albumin (BSA) by experimental and computational methods. Binding constant, binding site, mode of binding, and the BSA structural change upon AUR addition, were studied. UV-vis spectroscopy results and fluorescence quenching analysis proposed that AUR can form the ground state complex with BSA. Meantime, thermodynamic parameters (negative ΔH and ΔS values) revealed that hydrogen bonds and van der Waals interactions play major role, as intermolecular forces, in the AUR-BSA complex formation. Synchronous fluorescence spectra and circular dichroism (CD) data showed that the secondary structure of BSA did not change significantly in the presence of AUR. Moreover, molecular docking results showed that AUR binds to the subdomain IIIB of BSA.

A Sensitive, Simple and Direct Determination of Pantoprazole Based on a "Turn off-on" Fluorescence Nanosensor by Using Terbium-1,10-phenanthroline-silver Nanoparticles.

A new sensitive, simple, rapid, reliable and selective fluorometric method for the determination of pantoprazole (PAN) in human plasma and a pharmaceutical formulation has been developed. This technique is based on a quenching effect of silver nanoparticles (AgNPs) on the emission intensity of a fluorescent probe, terbium(III)-1,10-phenantroline (Tb(III)-phen) complex (due to a fluorescence resonance energy transfer (FRET) phenomenon between the Tb(III)-phen complex and AgNPs), and then restoring the fluorescence intensity of the Tb(III)-phen-AgNPs system upon the addition of PAN (turn off-on process). The effects of various factors on the proposed method including time, temperature, pH, order of the addition of various reagents and the concentration of AgNPs were investigated. Under the optimal conditions, a good linear relationship between the enhanced emission intensity of the Tb(III)-phen-AgNPs system and the PAN concentration was observed in the range of (10 - 1000) × 10-8 M. The limit of detection (LOD) and the limit of quantitation (LOQ) were 7.2 × 10-8 and 24.2 × 10-8 M, respectively. Also, the interferences of some common interfering species on the fluorescence intensity of the system were investigated. This simple and sensitive method was successfully applied for the determination of PAN in spiked human plasma samples and in its capsule formulation. The analytical recoveries were in the range of 88.54 - 101.33 and 90.07 - 98.85%, respectively.

A novel ultrasensitive and non-enzymatic "turn-on-off" fluorescence nanosensor for direct determination of glucose in the serum: As an alternative approach to the other optical and electrochemical methods.

A new, simple, rapid, highly sensitive and selective and non-enzymatic fluorometric method for direct determination of glucose in real samples was developed. The method was based on the inhibition of fluorescence resonance energy transfer (FRET) process between terbium (III)-1, 10-phenanthroline (Tb-phen) complex and silver nanoparticles (AgNPs). Upon the addition of glucose, the quenched FRET-based fluorescence of Tb-phen complex was gradually recovered by glucose via its strong adsorption on the surface of AgNPs and removal of Tb-phen complex from AgNPs surface. Therefore the fluorescence of Tb-phen complex switched to "turn-on" state. Under the optimum conditions, a linear relationship was obtained between the enhanced fluorescence intensity and glucose concentration in the range of (5-900) × 10-8 M with the detection limit of 1.94 × 10-8 M. The proposed sensing system was successfully applied to determine glucose in the spiked normal and diabetic patient serum samples after deproteinization with acetonitrile. Analytical recoveries from treated serum samples were in the range of 99.97-104.80% and 92.14-105.43%, respectively. The common interfering species, such as ascorbic acid, fructose and galactose did not cause interior interference due to unique emission properties of Tb-phen complex probe. Also the interaction of the Tb-phen complex with AgNPs, which led to the fluorescence intensity quenching of the complex, was further examined by FTIR technique. In short, as compared to most of the existing methods, the newly proposed method, provides some advantages and makes it promising for the direct rapid screening of glucose residues of real samples in clinical diagnosis of diabetes, as an alternative approach to the other exiting optical and electrochemical methods.

Multispectral and computational probing of the interactions between sitagliptin and serum albumin.

The binding of sitagliptin (SIT), an anti-diabetic drug, to human and bovine serum albumin (HSA and BSA; main serum transport proteins) was investigated using various spectroscopic and molecular docking techniques. The fluorescence data demonstrated that SIT quenched inherent fluorescence of these proteins through the formation of SIT-HSA/BSA complexes. The number of binding sites was obtained (~1) and binding constant (Kb) and effective quenching constant (Ka) were calculated as 104 for both systems. Based on thermodynamic parameters, the van der Waals forces and hydrogen bonding were the most important forces in the interactions between HSA/BSA and SIT, and the complex formation processes were spontaneous. The results of UV-vis absorption and FT-IR spectroscopic revealed that SIT induces small conformational changes in the structure of the proteins (HSA/BSA). The synchronous fluorescence (SF) spectroscopy demonstrated that the binding of SIT with HSA/BSA had no effect on the polarity around Trp and Tyr residues. The CD spectra showed changes in the secondary and tertiary structures of both proteins with a decrease in α-helices contents and an increase in ß-turn structures. The molecular docking and spectroscopic data verified the binding mechanisms between SIT and HSA/BSA, and revealed that SIT completely fits into the hydrophobic cavity between domain II and domain III of these proteins.

Determination of total phenols in tea infusions, tomato and apple juice by terbium sensitized fluorescence method as an alternative approach to the Folin-Ciocalteu spectrophotometric method.

A fast screening of total phenols in tea infusions, tomato and apple juice samples using terbium sensitized fluorescence is described. The proposed method is based on the fluorescence sensitization of terbium (Tb(3+)) by complexation with flavonols (quercein as a reference standard) (at pH 7.0), which fluoresces intensely with an emission maximum at 545nm when excited at 310nm. Quercetin and terbium cations (at pH 7.0) form a stable complex and the resulted emission at 545nm can be used for the determination of the total phenols concentration expressed in terms of "quercetin equivalent". Based on the obtained results, a sensitive, simple and rapid spectrofluorimetric method was developed for the determination of total phenols. In the optimum conditions, the calibration graph was linear from 0.01 to 2µgmL(-1), with the limit of detection of 0.002µgmL(-1). The relative standard deviation values were in the range of 0.75-2.3%. The total concentrations of quercetin equivalent in five tested samples were found in the range of 6.6-27.9µgmL(-1) and the results compare favorably with those obtained by spectrophotometric method (r=0.999).

Spectroscopic profiling and computational study of the binding of tschimgine: A natural monoterpene derivative, with calf thymus DNA.

DNA is a major target for a number of anticancer substances. Interaction studies between small molecules and DNA are essential for rational drug designing to influence main biological processes and also introducing new probes for the assay of DNA. Tschimgine (TMG) is a monoterpene derivative with anticancer properties. In the present study we tried to elucidate the interaction of TMG with calf thymus DNA (CT-DNA) using different spectroscopic methods. UV-visible absorption spectrophotometry, fluorescence and circular dichroism (CD) spectroscopies as well as molecular docking study revealed formation of complex between TMG and CT-DNA. Binding constant (Kb) between TMG and DNA was 2.27×104M-1, that is comparable to groove binding agents. The fluorescence spectroscopic data revealed that the quenching mechanism of fluorescence of TMG by CT-DNA is static quenching. Thermodynamic parameters (ΔH<0 and ΔS<0) at different temperatures indicated that van der Waals forces and hydrogen bonds were involved in the binding process of TMG with CT-DNA. Competitive binding assay with methylene blue (MB) and Hoechst 33258 using fluorescence spectroscopy displayed that TMG possibly binds to the minor groove of CT-DNA. These observations were further confirmed by CD spectral analysis, viscosity measurements and molecular docking.

Studies of interaction between terbium(III)-deferasirox and double helix DNA by spectral and electrochemical methods.

DNA binding studies of terbium(III)-deferasirox (Tb3+-DFX) complex were monitored to understand the reaction mechanism and introduce a new probe for the assay of DNA. In the present work, UV absorption spectrophotometry, fluorescence spectroscopy, circular dichroism (CD), cyclic voltammetry (CV) and viscosity measurement were employed to study the interactions of Tb3+-DFX with calf thymus DNA (ctDNA). The binding of Tb3+-DFX complex to ctDNA showed a hyperchromic effect in the absorption spectra and the increase in fluorescence quenching effect (amount) of Tb3+-DFX complex in the presence of ctDNA. The binding constants (Kb) for the complex with ctDNA were estimated to be 1.8×10(4) M(-1) through UV absorption spectrophotometry and fluorescence spectroscopy. Upon addition of the complex, clear decreases were observed in the viscosity of ctDNA. The CD spectra indicated that there are certain detectable conformational changes in the DNA double helix when the complex was added. The CV method showed that both anodic and cathodic peak potentials of Tb3+-DFX complex showed negative shifts on the addition of the ctDNA. Further, competitive methylene blue binding studies with fluorescence spectroscopy have shown that the complex can bind to ctDNA through nonintercalative mode. The experimental results suggest that Tb3+-DFX complex binds to DNA via groove binding and/or electrostatic binding mode.

Determination of methotrexate in biological fluids and a parenteral injection using terbium-sensitized method.

A new sensitive, simple and rapid method for determination of methotrexate (MTX) was developed based on quenching effects of MTX on the fluorescence intensity of Tb(3+)-1,10-phenanthroline complex. The fluorescence intensity was measured with excitation wavelength of 300 nm and emission wavelength of 545 nm and the quenched fluorescence intensity is proportional to the concentration of MTX in Tris-HCl buffer solution with a pH of 7.0. The effects of pH, time, order of addition of the reagents, temperature and the concentrations of Tb(3+), buffer and 1,10-phenanthroline were investigated and optimized. The obtained linear range for the determination of MTX was 0.02-10 µg/mL. The detection limits (signal: noise = 3) was 0.015 µg/mL and the relative standard deviation for replicated determinations of 1 µg/mL of MTX was 1.9%. The proposed method is a simple, practical and relatively free from interference effects and was successfully applied to assess MTX in urine, serum and samples of an injection solution.