Sorana Ionescu

Study of the interaction between ofloxacin and human serum albumin by spectroscopic methods

The binding of ofloxacin (OFLX) to human serum albumin (HSA) was investigated by fluorescence and circular dichroism (CD) techniques. The binding parameters have been evaluated by a fluorescence quenching method. Competitive binding measurements were performed in the presence of warfarin and ibuprofen and suggest binding to the warfarin site I of HSA. The distance r between donor (HSA) and acceptor (OFLX) was estimated according to the Forsters theory of non-radiatiative energy transfer. CD spectra revealed that the binding of OFLX to HSA induced conformational changes in HSA. Molecular docking was performed and shows that for the lowest energy complex OFLX is located in site I of HSA, which correlate to the competitive binding experiments.

Kaempferol-human serum albumin interaction: characterization of the induced chirality upon binding by experimental circular dichroism and TDDFT calculations.

The experimental induced circular dichroism (ICD) and absorption spectra of the achiral flavonoid kaempferol upon binding to human serum albumin (HSA) were correlated to electronic CD and UV-vis spectra theoretically predicted by time-dependent density functional theory (TDDFT). The neutral and four anionic species of kaempferol in various conformations were considered in the calculations. The appearance of the experimental ICD signal was rationalized in terms of kaempferol binding to HSA in a distorted, chiral, rigid conformation. The comparison between the experimental and simulated spectra allowed for the identification of the kaempferol species that binds to HSA, namely the anion generated by deprotonation of the hydroxyl group in position 7. This approach constitutes a convenient method for evidencing the binding species and for determining its conformation in the binding pocket of the protein. Its main advantage over the UV-vis absorption method lays in the fact that only the bound ligand species gives an ICD signal.

Photophysical Properties of Some Flavones Probes in Homogeneous Media

Photophysical properties of five hydroxyflavones (HF) (some typical models of flavonols), (3 - HF, 6 - HF, 7-HF, 3, 6 - diHF and 3, 7 - diHF) were studied in homogeneous media by means of UV–vis and steady-state and time resolved fluorescence spectroscopies. Their absorption and fluorescence characteristics based on the flavonols structure are presented and discussed. It was found that the fluorescence of the flavonols depends on the nature of the solvent and on their molecular structure, especially on the position and the number of the -OH groups of the substituted phenyl ring. Attention is paid to the number of the -OH groups that influence the excited-state intramolecular proton transfer (ESIPT) process. The fluorescence quantum yield and the lifetime of the flavonols in heterogeneous media have been also determined. The results are discussed with relevance to the flavonols as sensitive fluorescence probe and to their microenvironments in the systems of biological interest and especially in a typical protein environment.

Effect of pH on the fluorescence characteristics of some flavones probes.

The photophysical properties such as electronic absorption, molar extinction coefficient, emission spectra, fluorescence quantum yield and lifetime of three different hydroxyflavones (a typical model of flavonols) such as: 3-HF, 3,6-diHF and 3,7-diHF, have been studied in the pH range from 2.5 to 9.2. Both electronic absorption and fluorescence spectra are sensitive to pH. The fluorescence quantum yield at pH 7.4 of the mentioned flavones probes have been determined. The fluorescence lifetime of different emissive species (Normal, Tautomer and Anion forms) as pH dependence have been also estimated. The effect of pH on the intramolecular excited state proton transfer process (ESIPT) has been discussed. The normal and tautomeric forms change as a function of pH, the normal one being more sensitive. The position of the -OH group on the second aromatic ring in the flavonols structure has been also discussed. The results have relevance to compounds which have photoreactions accompanied by dual fluorescence.

Induced chirality in fisetin upon binding to serum albumin: experimental circular dichroism and TDDFT calculations

AbstractTheoretical absorption and electronic circular dichroism (ECD) spectra predicted via time-dependent density functional theory (TDDFT) calculations on the neutral and four anionic species of fisetin, an achiral flavonoid, were used to rationalize the experimental absorption and induced circular dichroism (ICD) spectra of the ligand upon binding to human serum albumin (HSA). On this basis, the mechanism responsible for the appearance of the ICD signal was ascribed to a distortion of the conformation of bound fisetin. Furthermore, comparison of the simulated and experimental spectra revealed that two fisetin species bind to HSA, namely, the neutral molecule and the anion deprotonated at the hydroxyl group in position 7, in a 1:1 ratio. The coupling of the theoretical results with the experimental absorption and ICD data allows identification of the flavonoid species that bind to the protein and evaluation of their conformation in the binding site. FigureReconvolution of the simulated ECD spectra of two fisetin species in different molar ratios

Characterization of a new norfloxacin metabolite monitored during a bioequivalence study by means of mass spectrometry and quantum computation.

A bioequivalence study of two formulations containing norfloxacin was used for identification and assay of the metabolite of norfloxacin in human serum samples. The bioequivalence study was based on an analytical method using liquid chromatography with fluorescence detection. The plasmatic profile of metabolite was similar to norfloxacin for both formulations. Three plasma fractions of norfloxacin metabolite from a volunteer were isolated by liquid chromatography and investigated by atmospheric-pressure chemical ionization mass-spectrometry. The structure of norfloxacin metabolite (7-aminoethylenamino-6-fluoro-4-hydroxy quinoline-3-carboxylic acid) was identified taking into account also the mass spectrometry investigations achieved for norfloxacin and ciprofloxacin (used as internal standard for the analytical method). A theoretical procedure based on quantum chemical calculations has been also used to explain the mass fragmentation in molecules of norfloxacin metabolite that differ from the molecule of norfloxacin.

Fluorescence Characteristics of some Flavones Probes in Different Micellar Media

The fluorescence characteristics of five hydroxiflavones (HFs) (some typical models of flavonols), (3 - HF, 6 - HF, 7-HF, 3, 6 - diHF and 3, 7-diHF) in the micellar media of non-ionic surfactant (Triton X-100), anionic surfactant (SDS) and the block copolymer Pluronic F127, have been investigated by means of UV–Vis and steady-state and time resolved fluorescence spectroscopies. Attention is paid to both excited-state intra-molecular proton transfer (ESIPT) as well as ground-state intermolecular proton transfer. The influence of the -OH groups as well as the effect of temperature on the dual fluorescence emission, the Normal and Tautomer emissions, are also investigated. The fluorescence quantum yield of the HFs in mentioned micellar media has been also determined. The results are discussed with relevance to the local environment of HFs as sensitive fluorescence probe in biological membrane systems.

Spectroscopic study of 3-Hydroxyflavone - protein interaction in lipidic bi-layers immobilized on silver nanoparticles.

The interaction of 3-Hydroxyflavone with serum proteins (BSA and HSA) in lecithin lipidic bi-layers (PC) immobilized on silver nanoparticles (SNPs), was studied by fluorescence and Raman spectroscopy. BSA secondary structure was quantified with a deconvolution algorithm, showing a decrease in α-helix structure when lipids were added to the solution. The effect of temperature on the rate of the excited-state intra-molecular proton transfer and on the dual fluorescence emission of 3-HF in the HSA/PC/SNPs systems was discussed. Evaluation of the antioxidant activity of 3-HF in HSA/PC/SNPs systems was also studied. The antioxidant activity of 3-HF decreased in the presence of SNPs. The results are discussed with relevance to the secondary structure of proteins and of the 3-HF based nano-systems to a topical formulation useful in the oxidative stress process.

Visible-light triggered photoswitching systems based on fluorescent azulenyl-substituted dithienylcyclopentenes

Novel azulenyl functionalized dithienylethenes were synthesized, and their photochromic reactivity, fluorescent and electrochemical properties were investigated. By tailoring the conventional dithienylcyclopentene chromophore with azulene groups, the photocyclization occurred with 405 nm wavelength light irradiation. These compounds exhibited relatively strong fluorescence at 524 nm when excited at 360 nm, which decreased along with the photocyclization. Furthermore, their electrochemical oxidation induced cyclization.

Organic co-crystals of 1,3-bis(4-pyridyl)azulene with a series of hydrogen-bond donors

The supramolecular interactions between 1,3-bis(4-pyridyl)azulene (azbbpy) as a hydrogen bond acceptor and a variety of aromatic hydrogen-bond donors produced new binary molecular co-crystals and a salt. All crystal structures display extensive and very complex hydrogen bond networks containing both homo- and heterosynthons formed through robust O–H⋯N and OH⋯O hydrogen bonds. The binary co-crystals of azbbpy with 4,4′-biphenol and 4,4′-oxo-bis-benzoic acid in a 1 : 1 ratio revealed that the recognition process differs. While 4,4′-biphenol acts both as a donor and an acceptor of protons, generating a two-dimensional supramolecular network in which azbbpy interacts with only one pyridine moiety, the co-crystallization with 4,4′-oxo-bis-benzoic acid yielded zig-zag chains formed by alternating acid–pyridine supramolecular heterosynthons. Co-crystallization with isophthalic acid yields a 2 : 1 system, 4, as hydrogen-bonded zig-zag supramolecular chains containing a R44(36) graph set organized through strong OH⋯N and weak C–H⋯N hydrogen bonds. Co-crystallization of azbbpy with trimesic acid in different ratios yielded two distinct systems: co-crystal 5 and salt 6. In 5, the trimesic acid molecules are assembled in supramolecular helical chains which are further connected by azbbpy through hydrogen bonding and π–π interactions into a 3D architecture with channels that host water molecules. A complicated supramolecular 3D architecture is also established between trimesic anions and azbbpyH+ cations in crystal 6, resulting in channels filled with disordered solvent molecules. These types of interactions were also evidenced in a quantitative way and confirmed on grounds of the Hirshfeld surface mapped with geometrical parameters that characterize close contacts in co-crystals. The optical properties of these supramolecular systems showed strong absorption bands in the visible region due to the azulene fragment.