Preparation of aminated core‐shell fluorescent nanoparticles and their application to the synchronous fluorescence determination of γ‐globulin

Amino‐modified silica nanoparticles (FSNPs) doped with fluorescein isothiocyanate (FITC) were synthesized by using an aqueous core of reverse‐micelle microemulsion as the nanoreactor in an easy one‐pot method. Due to the FITC conjugating with (3‐aminopropyl)triethoxysilane (APTS), the nanoparticles prevent the FITC from leaching from the silica matrix when immersed in aqueous solution. SEM, FTIR, fluorescence lifetime, a photobleaching experiment and synchronous fluorescence spectra were used to characterize the FSNPs. The synchronous fluorescence signal of FSNPs was enhanced when trace amounts of γ‐globulin (γ‐G) were added. Under the optimal experimental conditions, the enhanced fluorescence intensity (ΔF) was linear with the concentration of γ‐G (c) in the range 0.3–4.8 µg/mL, with a detection limit of 0.04 µg/mL. The proposed method is simple, sensitive for the determination of trace amounts of γ‐G and used to determine the content of γ‐G in synthetic samples with satisfactory results. Copyright © 2008 John Wiley & Sons, Ltd.     

A ‘turn‐on’ fluorescent chemosensor for quantification of serum albumin in aqueous solution at neutral pH

A fluorescent chemosensor 1 (4‐diethylamino‐2′‐hydroxychalcone) for detecting serum albumin with long‐wavelength emission, good selectivity and facile synthesis was reported. Upon the addition of bovine serum albumin (BSA) to an aqueous solution of 1 at neutral pH, a ‘turn‐on’ fluorescence response was observed at 596 nm based on a hydrophobic binding mode between 1 and BSA. A linear range of 0.10–1.00 mg/mL and a detection limit of 9.1 µg/mL for BSA were obtained, respectively. Moreover, 1 was successfully applied to detect BSA in real bovine serum samples with satisfied recovery and accuracy, which suggested that 1 could serve as a valid and effective fluorescent chemosensor for quantification of BSA. Copyright © 2015 John Wiley & Sons, Ltd.     

Study on the interactions of mapenterol with serum albumins using multi‐spectroscopy and molecular docking

The interactions of mapenterol with bovine serum albumin (BSA) and human serum albumin (HSA) have been investigated systematically using fluorescence spectroscopy, absorption spectroscopy, circular dichroism (CD) and molecular docking techniques. Mapenterol has a strong ability to quench the intrinsic fluorescence of BSA and HSA through static quenching procedures. At 291 K, the binding constants, K_a, were 1.93 × 10³ and 2.73 × 10³ L/mol for mapenterol–BSA and mapenterol–HAS, respectively. Electrostatic forces and hydrophobic interactions played important roles in stabilizing the mapenterol–BSA/has complex. Using site marker competitive studies, mapenterol was found to bind at Sudlow site I on BSA/HSA. There was little effect of K⁺, Ca²⁺, Cu²⁺, Zn²⁺ and Fe³⁺ on the binding. The conformation of BSA/HSA was changed by mapenterol, as seen from the synchronous fluorescence spectra. The CD spectra showed that the binding of mapenterol to BSA/HSA changed the secondary structure of BSA/HSA. Molecular docking further confirmed that mapenterol could bind to Sudlow site I of BSA/HSA. According to Förster non‐radiative energy transfer theory (FRET), the distances r₀ between the donor and acceptor were calculated as 3.18 and 2.75 nm for mapenterol–BSA and mapenterol–HAS, respectively. Copyright © 2015 John Wiley & Sons, Ltd.     

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 (K_b) 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.     

Chemiluminescence determination of promazine in human serum and drug formulations using Ru(phen)32+–Ce(IV) system and a chemometrical optimization approach

A new chemiluminescence (CL) method using flow injection has been described for the rapid and sensitive determination of promazine hydrochloride (PMH). The method is based on the CL reaction of PMH with tris(1,10 phenanthroline)ruthenium(II), [Ru(phen)₃²⁺] and Ce(IV) in sulfuric acid medium. Effects of chemical variables were investigated employing central composite design and response surface methodology. Under the optimum conditions, the CL intensity was proportional to the concentration of the drug in solution over the ranges 0.020–0.32 and 0.32–32 µg/mL. The limit of detection (signal‐to‐noise ratio = 3) was 0.012 µg/mL. The method was applied successfully to the determination of PMH in drug formulations and human serum (recovery percentages between 96.7 and 105.0%). The relative standard deviation for 11 replicate determinations of 1.5 µg/mL of PMH was 1.7%. The minimum sampling rate was 100 samples per hour. Copyright © 2009 John Wiley & Sons, Ltd.     

Flow‐injection analysis for meloxicam based on tris(2,2′‐bipyridine) ruthenium(II)–Ce(IV) chemiluminescent system

It was found that meloxicam could enhance the chemiluminescence (CL) of the tris(2,2'‐bipyridine) ruthenium(II)–Ce(IV) system in the medium of sulfate acid. Based on this phenomenon a new flow‐injection system with chemiluminescent detection has been proposed for determination of meloxicam. Under optimum conditions, meloxicam had a good linear relationship with the CL intensity in the concentration range of 6.0  10^?4 to 1.0 µg/mL and the detection limit was 3.7 × 10^?4 µg/mL. The proposed method was applied to detect meloxicam in tablets and a satisfactory recovery was obtained. The possible mechanism for this CL system is also discussed in this paper. Copyright © 2009 John Wiley & Sons, Ltd.     

Synthesis and study on the binding of thiazol‐2(3H)‐ylidine derivative with human serum albumin using spectroscopic and molecular docking methods

In this article, a facile and convenient synthesis of thiazol‐2(3H)‐ylidine derivatives of fatty acid ( 3a – c ) is described. The binding of N′‐(4,5‐dimethyl‐3‐penylthiazol‐2(3H)‐ylidine)octadec‐9‐enehydrazide ( 3a ) with human serum albumin (HSA) is explored using various spectral methods and molecular docking. Fluorescence quenching results show that 3a induces conformational changes in HSA and the polarity around the tryptophan residues is increased. Stern–Volmer quenching plots at different temperatures (298, 305 and 312 K) show that the fluorescence quenching mechanism is static quenching. Synchronous fluorescence, 3D fluorescence spectra, circular dichroism and Fourier transform infrared spectroscopy are used to determine the structural change in HSA on interaction with 3a . Förster resonance energy transfer analysis shows that the binding distance (r₀ = 2.78 nm) between HSA (Trp214) and 3a is within the of range 2–8 nm for quenching to occur. The molecular docking study also confirms that 3a is located in subdomain IIA (site I) of HSA and is stabilized by hydrogen bonding and hydrophobic forces.     

A validated HPLC method for the determination of dimethyl‐4,4′‐dimethoxy‐5,6,5′,6′‐dimethylene dioxybiphenyl‐2,2′‐dicarboxylate (DDB) with fluorescence detection in raw material and pill form: application to an in vitro dissolution test and a content uniformity test

A simple, sensitive and rapid HPLC method with fluorescence detection for the determination of dimethyl‐4,4′‐dimethoxy‐5,6,5′,6′‐dimethylene dioxybiphenyl‐2,2′‐dicarboxylate (DDB) in the raw material and pill form was developed. Liquid chromatography was performed on a C₁₈ column (250 × 4.6 mm i.d., 5 µm particle size), the mobile phase consisted of methanol and 0.05 M sodium dihydrogen phosphate buffer (80 : 20, v/v), and the apparent pH of the mobile phase was adjusted to 3. The fluorescence detector was operated at excitation/emission wavelengths of 275/400 nm. The proposed method allows the determination of DDB within concentration range 0.1–1.5 µg/mL with a limit of detection of 0.032 µg/mL, a limit of quantification of 0.097 µg/mL and a correlation coefficient of 0.9997. The proposed method has been successfully applied for the analysis of DDB in its pills with a percentage recovery of 98.45 ± 0.32. The method was fully validated according to ICH guidelines. Moreover, the high sensitivity of the method permits its use in an in vitro dissolution test for DDB under simulated intestinal conditions. In addition, the proposed method was extended to a content uniformity test according to USP guidelines. Copyright © 2014 John Wiley & Sons, Ltd.     

Trace analysis of N‐acetyl‐L‐cysteine using luminol–H2O2 chemiluminescence system catalyzed by silver nanoparticles

N‐Acetyl‐L‐cysteine (NAC) can inhibit the luminol–H₂O₂, reaction, which is catalyzed by silver nanoparticles. Based on this phenomenon a new method was developed for NAC determination. Under optimum conditions, a linear relationship between chemiluminescence intensity and NAC concentration was found in the range 0.034–0.98 µg/mL. The detection limit was 0.010 µg/mL (S/N =3), and the relative standard deviation (RSD) was <5% for 0.480 µg/mL NAC (n =5). This simple, sensitive and inexpensive method has been applied to measure the concentration of NAC in pharmaceutical tablets. Copyright © 2014 John Wiley & Sons, Ltd.     

Validated spectrofluorimetric method for the determination of carbamazepine in pharmaceutical dosage forms after reaction with 4‐chloro‐7‐‐nitrobenzo‐2‐oxa‐1,3‐diazole (NBD‐Cl)

A sensitive and simple spectrofluorimetric method has been developed and validated for the determination of the anti‐epileptic drug carbamazepine (CBZ) in its dosage forms. The method was based on a nucleophilic substitution reaction of CBZ with 4‐chloro‐7‐nitrobenzo‐2‐ oxa‐1,3‐diazole (NBD‐Cl) in borate buffer (pH 9) to form a highly fluorescent derivative that was measured at 530 nm after excitation at 460 nm. Factors affecting the formation of the reaction product were studied and optimized, and the reaction mechanism was postulated. The fluorescence–concentration plot is rectilinear over the range of 0.6–8 µg/mL with limit of detection of 0.06 µg/mL and limit of quantitation of 0.19 µg/mL. The method was applied to the analysis of commercial tablets and the results were in good agreement with those obtained using the reference method. Validation of the analytical procedures was evaluated according to ICH guidelines. Copyright © 2015 John Wiley & Sons, Ltd.     

Enantiospecific Analysis of 8‐Prenylnaringenin in Biological Fluids by Liquid‐Chromatography‐Electrospray Ionization Mass Spectrometry: Application to Preclinical Pharmacokinetic Investigations

8‐Prenylnaringenin (8PN) is a naturally occurring bioactive chiral prenylflavonoid found most commonly in the female flowers of hops (Humulus lupulus L.). A stereospecific method of analysis for 8PN in biological fluids is necessary to study the pharmacokinetic disposition of each enantiomer. A novel and simple liquid chromatographic‐electrospray ionization‐mass spectrometry (LC‐ESI‐MS) method was developed for the simultaneous determination of R‐ and S‐8PN in rat serum and urine. Carbamazepine was used as the internal standard (IS). Enantiomeric resolution of 8PN was achieved on a Chiralpak^® AD‐RH column with an isocratic mobile phase consisting of 2‐propanol and 10 mM ammonium formate (pH 8.5) (40:60, v/v) and a flow rate of 0.7 mL/min. Detection was achieved using negative selective ion monitoring (SIM) of 8PN at m/z 339.15 for both enantiomers and positive SIM m/z at 237.15 for the IS. The calibration curves for urine were linear over a range of 0.01–75 µg/mL and 0.05–75 µg/mL for serum with a limit of quantification of 0.05 µg/mL in serum and 0.01 µg/mL in urine. The method was successfully validated showing that it was sensitive, reproducible, and accurate for enantiospecific quantification of 8PN in biological matrices. The assay was successfully applied to a preliminary study of 8PN enantiomers in rat. Chirality 26:419–426, 2014. © 2014 Wiley Periodicals, Inc.     

Study of the enhancement of a new chemiluminescence reaction and its application to determination of β‐lactam antibiotics

An enhanced thiosemicarbazide(TSC)–H₂O₂ chemiluminescence (CL) system was established and proposed as a new analytical method for determination of β‐lactam antibiotics, ampicillin sodium and amoxicillin at microgram levels. The method is based on the inhibition of CL emission accompanying oxidation of TSC by H₂O₂ in alkaline medium. The effect of anionic, cationic, and non‐ionic surfactants on the CL emission of the system was studied. Both N‐cetyl‐N,N,N‐trimethylammonium bromide (CTMAB) and Triton X‐100, unlike sodium dodecyl sulfate (SDS), reinforced the CL intensity and were efficient to approximately the same level. The effect of the presence of eight non‐aqueous solvents on the CL system was also investigated. Upon addition of both of the non‐ionic surfactant, Triton X‐100, and the non‐aqueous solvent, N,N‐dimethyl formamide (DMF), the intensity of the CL reaction was increased 100‐fold. This method allows the measurement of 25–545 µg amoxicillin, and 35–350 µg ampicillin sodium. The detection limits are 8 µg for amoxicillin and 9 µg for ampicillin sodium. The relative standard deviations of six replicate measurements of 200 µg amoxicillin and 200 µg ampicillin sodium were 1.9 and 2.1%, respectively. The effect of foreign species on the determination of amoxicillin and ampicillin sodium was also examined. The proposed method was successfully applied to the determination of ampicillin sodium and amoxicillin in some pharmaceutical dosage forms. Copyright © 2008 John Wiley & Sons, Ltd.     

Flow‐injection chemiluminescent determination of estrogen benzoate using the tris(1,10‐phenanthroline) ruthenium(II)–permanganate system

Chemiluminescence (CL) detection for the determination of estrogen benzoate, using the reaction of tris(1,10–phenanthroline)ruthenium(II)–Na₂SO₃–permanganate, is described. This method is based on the CL reaction of estrogen benzoate (EB) with acidic potassium permanganate and tris(1,10–phenanthroline)ruthenium(II). The CL intensity is greatly enhanced when Na₂SO₃ is added. After optimization of the different experimental parameters, a calibration graph for estrogen benzoate is linear in the range 0.05–10 µg/mL. The 3 s limit of detection is 0.024 µg/mL and the relative standard deviation was 1.3% for 1.0 µg/mL estrogen benzoate (n = 11). This proposed method was successfully applied to commercial injection samples and emulsion cosmetics. The mechanism of CL reaction was also studied. Copyright © 2011 John Wiley & Sons, Ltd.     

Study on the interaction between palladium(II)–lincomycin chelate and erythosine by absorption,fluorescence and resonance Rayleigh scattering spectra and its analytical applications

In pH 5.0–5.4 HAc–NaAc buffer solution, lincomycin (Linco) reacted with Pd(II) to form 1:1 cationic chelate, which could further react with erythrosine (Ery) to form 1:1 ion‐association complexes (Pd–Linco)Ery. As a result, not only were the absorption and fluorescence spectra changed, but also the resonance Rayleigh scattering (RRS) intensity was greatly enhanced. These phenomena offered useful means for the determination of Linco by spectrophotometry, fluorescence and RRS methods. The linear range and detection limit of Linco were 0.20–3.00 µg/mL and 0.057 µg/mL, 0.20–4.80 µg/mL and 0.061 µg/mL, 0.05–2.70 µg/mL and 0.015 µg/mL for the spectrophotometric, fluorescence quenching and RRS methods, respectively. Among these, the RRS method obtained the highest sensitivity. Therefore, the optimum reaction conditions and the influences of coexisting substances were investigated using the RRS method. A simple, sensitive and rapid method has been developed for the determination of Linco in either the pharmaceutical form or human body fluids, and the reasons for RRS enhancement are discussed. Copyright © 2008 John Wiley & Sons, Ltd.     

6‐O‐(3″, 4″‐di‐O‐trans‐cinnamoyl)‐α‐l‐rhamnopyranosylcatalpol and verbascoside: Cytotoxicity,cell cycle kinetics,apoptosis, and ROS production evaluation in tumor cells

The aim of this study was to evaluate the impact that 6‐O‐(3″, 4″‐di‐O‐trans‐cinnamoyl)‐α‐ l ‐rhamnopyranosylcatalpol (Dicinn) and verbascoside (Verb), two compounds simultaneously reported in Verbascum ovalifolium, have on tumor cell viability, apoptosis, cell cycle kinetics, and intracellular reactive oxygen species (ROS) level. At 100 µg/mL and 48 hours incubation time, Dicinn and Verb produced good cytotoxic effects in A549, HT‐29, and MCF‐7 cells. Dicinn induced cell‐cycle arrest at the G₀/G₁ phase and apoptosis, whereas Verb increased the population of subG₁ cells and cell apoptosis rates. Furthermore, the two compounds exhibited time‐dependent ROS generating effects in tumor cells (1‐24 hours). Importantly, no cytotoxic effects were induced in nontumor MCF‐10A cells by the two compounds up to 100 µg/mL. Overall, the effects exhibited by Verb in tumor cells were more potent, which can be correlated with its structural features, such as the presence of phenolic hydroxyl groups.     

Chemiluminescence method for the determination of piroxicam by the enhancement of the tris‐(4,7‐diphenyl‐1,10‐phenanthrolinedisulphonic acid) ruthenium(II) (RuBPS)–cerium(IV) system and its application

A simple, rapid chemiluminescence (CL) method was described for the determination of piroxicam, a commonly used analgesic agent drug. A strong CL signal was detected when cerium(IV) sulphate was injected into tris‐(4,7‐diphenyl‐1,10‐phenanthrolinedisulphonic acid) ruthenium(II) (RuBPS)–piroxicam solution. The CL signal was proportional to the concentration of piroxicam in the range 2.8 × 10^–8–1.2 × 10^–5 mol/L. The detection limit was 2 × 10^–8 mol/L and the relative standard deviation (RSD) was 3.7% (c = 7.0 × 10^–7 mol/L piroxicam; n = 11). The proposed method was applied to the determination of piroxicam in pharmaceutical preparations in capsules, spiked serum and urine samples with satisfactory results. Copyright © 2008 John Wiley & Sons, Ltd.     

Characterization of intermolecular interaction between cyanidin‐3‐glucoside and bovine serum albumin: Spectroscopic and molecular docking methods

The intermolecular interaction between cyanidin‐3‐glucoside (Cy‐3‐G) and bovine serum albumin (BSA) was investigated using fluorescence, circular dichroism and molecular docking methods. The experimental results revealed that the fluorescence quenching of BSA at 338 nm by Cy‐3‐G resulted from the formation of Cy‐3‐G–BSA complex. The number of binding sites (n) for Cy‐3‐G binding on BSA was approximately equal to 1. The experimental and molecular docking results revealed that after binding Cy‐3‐G to BSA, Cy‐3‐G is closer to the Tyr residue than the Trp residue, the secondary structure of BSA almost not change, the binding process of Cy‐3‐G with BSA is spontaneous, and Cy‐3‐G can be inserted into the hydrophobic cavity of BSA (site II′) in the binding process of Cy‐3‐G with BSA. Moreover, based on the sign and magnitude of the enthalpy and entropy changes (ΔH⁰ = – 29.64 kcal/mol and ΔS⁰ = – 69.51 cal/mol K) and the molecular docking results, it can be suggested that the main interaction forces of Cy‐3‐G with BSA are Van der Waals and hydrogen bonding interactions. Copyright © 2013 John Wiley & Sons, Ltd.     

Resonance light‐scattering enhancement effect of the protein–Y3+–TTA–SLS system and its analytical application

In this paper, a sensitive resonance light scattering (RLS) method for the determination of protein is reported. In the Tris–HCl (pH 7.50) buffer, protein enhanced the RLS intensity of the Y³⁺–2‐thenoyltrifluoroacetone (TTA)–sodium dodecyl sulphate (SLS) system. The enhanced RLS intensities were in proportion to the concentrations of proteins in the range 8.0 × 10^?9–1.0 × 10^?5 g/mL for BSA, 1.0 × 10^–8–1.0 × 10^?5 g/mL for HSA and 1.0 × 10^–8–1.0 × 10^?6 g/mL for EA, and their detection limits were 5.0, 5.4 and 6.7 ng/mL, respectively. Actual samples were satisfactorily determined. The interaction mechanism was also studied. Copyright © 2008 John Wiley & Sons, Ltd.     

A study on the interaction between 3‐spiro‐piperidones and bovine serum albumin using spectroscopic approaches

The interaction between 3‐spiro‐2′‐pyrrolidine‐3′‐spiro‐3″‐piperidine‐2,3″‐dione (PPD) and bovine serum albumin (BSA) in aqueous solution was studied using fluorescence and UV–vis spectroscopy. Fluorescence emission data revealed that BSA (1.00 × 10^‐5 mol/L) fluorescence was statically quenched by PPD at various concentrations, which implies that a PPD–BSA complex was formed. The binding constant (K_A), the number of binding sites (n) and the specific binding site of the PPD with BSA were determined. Energy‐transfer efficiency parameters were determined and the mechanism of the interaction discussed. The thermodynamic parameters, ΔG, ΔH and ΔS, were obtained according to van't Hoff's equation, showing the involvement of hydrophobic forces in these interactions. The effect of PPD acting on the BSA conformation was detected by synchronous fluorescence. Copyright © 2012 John Wiley & Sons, Ltd.