Microcavity-Supported Lipid Bilayers; Evaluation of Drug-Lipid Membrane Interactions by Electrochemical Impedance and Fluorescence Correlation Spectroscopy.

Many drugs have intracellular or membrane-associated targets, thus understanding their interaction with the cell membrane is of value in drug development. Cell-free tools used to predict membrane interactions should replicate the molecular organization of the membrane. Microcavity array-supported lipid bilayer (MSLB) platforms are versatile biophysical models of the cell membrane that combine liposome-like membrane fluidity with stability and addressability. We used an MSLB herein to interrogate drug-membrane interactions across seven drugs from different classes, including nonsteroidal anti-inflammatories: ibuprofen (Ibu) and diclofenac (Dic); antibiotics: rifampicin (Rif), levofloxacin (Levo), and pefloxacin (Pef); and bisphosphonates: alendronate (Ale) and clodronate (Clo). Fluorescence lifetime correlation spectroscopy (FLCS) and electrochemical impedance spectroscopy (EIS) were used to evaluate the impact of drug on 1,2-dioleyl- sn-glycerophosphocholine and binary bilayers over physiologically relevant drug concentrations. Although FLCS data revealed Ibu, Levo, Pef, Ale, and Clo had no impact on lipid lateral mobility, EIS, which is more sensitive to membrane structural change, indicated modest but significant decreases to membrane resistivity consistent with adsorption but weak penetration of drugs at the membrane. Ale and Clo, evaluated at pH 5.25, did not impact the impedance of the membrane except at concentrations exceeding 4 mM. Conversely, Dic and Rif dramatically altered bilayer fluidity, suggesting their translocation through the bilayer, and EIS data showed that resistivity of the membrane decreased substantially with increasing drug concentration. Capacitance changes to the bilayer in most cases were insignificant. Using a Langmuir-Freundlich model to fit the EIS data, we propose Rsat as an empirical value that reflects permeation. Overall, the data indicate that Ibu, Levo, and Pef adsorb at the interface of the lipid membrane but Dic and Rif interact strongly, permeating the membrane core modifying the water/ion permeability of the bilayer structure. These observations are discussed in the context of previously reported data on drug permeability and log P.

Production and characterization of a monoclonal antibody for Pefloxacin and mechanism study of antibody recognition.

In this report, an artificial antigen (PFLX-BSA: Pefloxacin connected bovine serum albumin) was successfully prepared. The monoclonal antibody against pefloxacin was produced and characterized using a direct competitive ELISA. The linear range of detection was 0.115-6.564 µg/L. The limit of detection defined as IC15 was 0.170 ± 0.05 µg/L and the IC50 was 0.902 ± 0.03 µg/L. The antibody variable region genes were amplified, assembled, and sequenced. A three-dimensional structural model of the variable region was constructed to study the mechanism of antibody recognition using molecular docking analysis. Three predicted essential amino acids, Thr53, Arg97 of heavy chain and Thr52 of light chain, were mutated to verify the theoretical model. Three mutants lost binding activity significantly against pefloxacin as predicted. These may provide useful insights for studying antigen-antibody interaction mechanisms to improve antibody affinity maturation in vitro.

Pharmacokinetics and tissue distribution of pefloxacin mesylate in chickens.

A specific, sensitive and stable high-performance liquid chromatography (HPLC)-based analytical method was established to determine the level of pefloxacin mesylate (PM) in the plasma and various tissues of chickens. Chickens were randomly assigned to 12 equal experiment groups, including 11 treatment groups and one control group. The chickens in the treatment groups received oral administration of PM and were sacrificed at different pre-determined time points, with their blood and various organs harvested, extracted and analyzed by HPLC to quantify the level of the residual antibiotic. Method validation studies indicated that the HPLC measurement showed excellent precision, reproducibility, stability and robustness. The obtained pharmacokinetic parameters suggested that PM reached peak levels in various tissues within 1-2 h after its oral administration, and was mainly concentrated in liver and kidney. The antibiotic was also found to be cleared from chicken crureus, brain, testes, ovaries and pancreas at higher rates compared with other organs. Overall, the rapid accumulation of PM could at least be partially attributed to its relatively slow organ clearance. These results could serve as a useful guidance for the rational use of PM and other quinolone-derived antimicrobials in the treatment of infectious diseases in chickens and other animals.

Comparison between Radioanalysis and 19F Nuclear Magnetic Resonance Spectroscopy in the Determination of Mass Balance, Metabolism, and Distribution of Pefloxacin.

Mass balance and metabolism studies using radiolabeled substances are well recognized as an important part of the drug development process. In this study, we directly assessed the use of fluorine nuclear magnetic resonance (19F NMR) to achieve quantitative mass balance, metabolism, and distribution information for fluorinated compounds, without the need for radiolabeled synthesis or study. As a test case, the disposition of pefloxacin, a fluoroquinolone antibiotic, was evaluated in rats using quantitative 19F NMR in parallel with a radiolabeled study. Urine, bile, and feces samples were collected over specific periods after oral administration of either 25 mg/kg [14C]pefloxacin or 25 mg/kg pefloxacin and were subsequently profiled by radioactivity or 19F NMR, respectively. The percentage of dose excreted in each matrix was comparable between the two methods, with the total dose recovered by radioactivity and 19F NMR determined to be 86.8% and 81.8%, respectively. In addition, plasma samples were collected to determine the exposure of pefloxacin and its circulating metabolites. The plasma exposure of pefloxacin determined by 19F NMR was within 5% to that calculated by a validated liquid chromatography-tandem mass spectrometry bioanalytical method. By both methods, pefloxacin was identified as the major circulating entity, with pefloxacin glucuronide as the major circulating metabolite. Quantitative analysis of metabolites in excreta was generally comparable between the two methods. In selected tissues, both methods indicated that the parent drug accounted for most of the drug-related material. In summary, we have demonstrated that 19F NMR can be used as an alternative method to conventional radiolabeled studies for compounds containing fluorine without the need for radiolabeled synthesis/study.

Study of a water-soluble fluorescent sensor based on the Eu(III) pefloxacin complex.

The antibiotic type organic structure pefloxacin binds well with europium (III) ions as a useful scaffold for assembling optical probes and allows energy transfer from ligand to metal ions through coordination linkages. This water-soluble chemosensor demonstrated significant 'off-on (red)' changes from an alkaline to a neutral environment (pH 14-8). The emission changed from red to blue under acidic conditions (pH 7-2). The whole process was completely reversible and effective within the pH range 2 to 14. Moreover, this probe system exhibited distinct luminescence quenching upon the addition of Cu2+ or Fe3+ . This general modular route will permit easy detection and the concept can be extended to a variety of quinolones for sensing purposes.

Study of silver nanoparticles sensitized fluorescence and second-order scattering of terbium(III)-pefloxacin mesylate complex and determination of pefloxacin mesylate.

α-Keto acid of pefloxacin mesylate (PFLX) can form the complex with Terbium(III). The intramolecular energy from PFLX to Terbium(III) ion takes place when excited, and thus Terbium(III) excited state is formed and then emits the characteristic fluorescence of Terbium(III), locating at 490, 545, 580, and 620 nm. The second-order scattering (SOS) peak at 545 nm also appears for the complex with the exciting wavelength of 273 nm. When the silver nanoparticles are added to the system, the luminescence intensity at 545 nm greatly increased. So, with the adding of nanoparticles to the Terbium(III)-PFLX complex, not only is the intramolecular energy promoted but also the SOS intensity is enhanced. The experimental results show that it is the silver nanoparticles with certain size and certain concentration which can greatly enhance the fluorescence-SOS intensity, and the relative intensity at 545 nm is proportional to the amount of PFLX. Based on this phenomenon, a novel method for the determination of PFLX has been developed and applied to the determination of PFLX in capsule and serum samples.

Experimental and theoretical studies on the molecular properties of ciprofloxacin, norfloxacin, pefloxacin, sparfloxacin, and gatifloxacin in determining bioavailability.

The aim of this investigation is to identify, by in silico and in vitro methods, the molecular determinants, e.g., solubility in an aqueous medium and lipophilic properties, which have an effect on the bioavailability of five selected fluoroquinolones. These properties were estimated by analysis of the electrostatic potential pattern and values of free energy of solvation as well as the partition coefficients of the studied compounds. The study is based on theoretical quantum-chemical methods and a simple experimental shake-flask technique with two immiscible phases, n-octanol and phosphate buffer. The solvation free energy values of compounds in both environments appeared to be negative. The wide range of electrostatic potential from negative to positive demonstrates the presence of dipole-dipole intermolecular interactions, while the high electron density at various sites indicates the possibility of hydrogen bond formation with solvent molecules. High partition coefficient values, obtained by summing the atomic contributions, did not take various correction factors into account and therefore were not accurate. Theoretical partition coefficient values based on more accurate algorithms, which included these correction factors (fragmental methods), yielded more accurate values. Theoretical methods are useful tools for predicting the bioavailability of fluoroquinolones.

Electro-Fenton degradation of pefloxacin using MOFs derived Cu, N co-doped carbon as a nanocomposite catalyst.

Electro-Fenton (EF) can in-situ produce H2O2 and effectively activate H2O2 to generate powerful reactive species for the destruction of contaminants under acidic conditions, however, the production of iron-containing sludge and requirement of low working pH significantly hinder its practical application. Herein, a novel Cu, N co-doped carbon (Cu-N@C) with metal organic framework (MOF) as a precursor was constructed and adopted for the elimination of pefloxacin (PEF) in the heterogeneous electro-Fenton (HEF) process. PEF could be almost completely removed within 1 h and total organic carbon (TOC) removal efficiency was 48.57% within 6 h. Meanwhile, Cu-N@C had good repeatability and environmental adaptability, it can still maintain excellent catalytic performance after 10 cycles, and it exhibited satisfactory remediation performance in simulated water matrix. In addition, the HEF process catalyzed by Cu-N@C also showed satisfactory degradation effect on other organic pollutants including atrazine, methylene blue, and chlorotetracycline. Under the action of impressed current, the HEF system could generate H2O2 in-situ, and the active species could be generated in the redox cycle of Cu0/Cu1+/Cu2+. Electron paramagnetic resonance and quenching experiments confirmed that •OH was the dominant active species in the degradation of organic compounds. The degradation process of PEF was studied by mass spectrometry analysis of intermediate products. This study provided a simple method to prepare MOF-based electrocatalyst, which exhibits promising application potential for treatment wastewater.

Ion coordination and chelation in Eu-MOFs matrices: Ultrafast fluorescence visual quantification monitoring of antibiotic residues.

Rapid monitoring of trace antibiotics in the field in real time is essential for environment forewarning and human health. High sensitivity and real-time on-site quantitative monitoring of antibiotic residues can be accomplished by integrating portable sensors alongside fluorescent optics to construct an intelligent sensing platform that smoothly eliminates the instability of conventional detection methods. In this study, a ratiometric fluorescence sensor for the ultrasensitive detection of pefloxacin was built employing the photoinduced electron transfer (PET) mechanism from red Eu-MOFs to Mn2+-PEF complex. A visual color change results from the photoinduced electron transfer process from manganese ions to pefloxacin weakening the ligand metal charge transfer (LMCT) process in Eu-MOFs. This enables the ultrafast visible detection of pefloxacin and produces a transient shift in visual color with a detection limit as low as 15.4 nM. For the detection of pefloxacin in water, tomato, and raw pork samples, various sensing devices based on the developed fluorescent probes exhibit good practicability and accuracy. With the development of the ratiometric fluorescence sensing probe, it is now possible to quickly and quantitatively identify pefloxacin residues in the environment, offering a new method for ensuring the safety of food and people's health.

[Molecular transport mechanism of pefloxacin mesylate binding with transferrin].

The binding mechanism between pefloxacin mesylate (PM) and transferrin (Tf) was explored using spectral experiment combined with molecular modeling techniques. The binding parameters and thermodynamic functions of PM-Tf solution system were measured at different temperatures. The effect of PM on molecular conformation of Tf was investigated and the interaction mechanism was also discussed. The results showed that dynamic quenching mechanism occurs with PM binding to Tf. The value of binding distances (r) is low, which indicates the occurrence of energy transfer. The drug had conformational effect on Tf, which resulted in changes of hydrophobic environment of the binding domain in Tf. According to the obtained thermodynamic parameters, the main interaction force between PM and Tf is attributed to hydrophobic bonding. The results of molecular modeling revealed that hydrophobic and hydrogen bonds are main binding forces in the PM-Tf system. These results were in accordance with spectral experiments. The research results have given a better theoretical reference for the study of pharmacological mechanism between protein and quinolone.

Third generation fluoroquinolones antibacterial drug based mixed-ligand Cu(II) complexes: structure, antibacterial activity, superoxide dismutase activity and DNA-interaction approach.

The copper(II) complexes of the type [Cu(SPF)(A(n))Cl]/[Cu(PFL)(A(n))Cl] (where SPF is sparfloxacin, PFL is pefloxacin and A(n) is 2,2'-dipyridylamine/pyridine-2-carboxalehyde/thiophene-2-carboxaldehyde) were synthesised and were found to have a pyramidal geometry with a square base. The superoxide dismutase (SOD) like activity of the complexes were measured using an NBT/NADH/PMS system, these were expressed in terms of the concentration of complex which termianates the formation of formazan by 50% (IC50 value) and found to range from 0.781 to 1.354 µM. The interactions of the complexes with DNA were studied by absorption titration, viscosity measurement and gel electrophoresis under physiological conditions. The antimicrobial efficiency of the complexes were tested on five different microorganisms and showed good biological activity.

Solvothermal synthesis of InNbO4 cubes for efficient degradation of pefloxacin.

A novel solvothermal process for synthesizing InNbO4 nanomaterials was developed. In this manner, a series of InNbO4 samples was synthesized. It was shown that reaction temperature and precursor pH had strong influence on the attributes of InNbO4 samples. The X-ray diffraction patterns revealed that all the samples possessed monoclinic structure and the optimal reaction condition was found at 250 °C with a pH of 5. Scanning electron microscopy images of different InNbO4 samples showed various morphologies. Transmission electron microscopy verified the synthesized InNbO4-pH 5 was single-crystal cubes. X-ray photoelectron spectra verified the existence of In, Nb, and O in InNbO4-pH 5 sample. The band gap of InNbO4-pH 5 was calculated to be 2.51 eV. The photocurrent intensity of InNbO4-pH 5 was the highest among the prepared samples. The photocatalytic degradation of pefloxacin was investigated using these samples. The InNbO4-pH 5 exhibited best degradation efficiency among these samples. The removal efficiency of pefloxacin with InNbO4-pH 5 could reach 80.2% in 60 min. Based on free radical capture results, superoxide radicals and holes showed to be the dominant active species. In addition, UHPLC/MS/MS was used to identify the degradation intermediates. Five new pefloxacin degradation products were found and possible degradation pathways were suggested.

Photolysis of enrofloxacin, pefloxacin and sulfaquinoxaline in aqueous solution by UV/H2O2, UV/Fe(II), and UV/H2O2/Fe(II) and the toxicity of the final reaction solutions on zebrafish embryos.

In this work, the photolysis of enrofloxacin (ENR), pefloxacin (PEF), and sulfaquinoxaline (SQX) in aqueous solution by UV combined with H2O2 or ferrous ions (Fe(II)), as well as Fenton (Fe(II)/H2O2) processes, was investigated. In addition, the toxicity of the final reaction solution after UV/H2O2/Fe(II) treatment toward zebrafish embryos was determined. The degradation of the test compounds followed pseudo-first-order reaction kinetics. The optimum concentrations of H2O2 for ENR, PEF and SQX removal under UV/H2O2 treatment were 20, 20 and 5 mM, respectively. The optimum concentrations of Fe(II) for ENR, PEF and SQX removal in the UV/Fe(II) system were 0.25, 10, and 1 mM, respectively. For the UV/H2O2/Fe(II) system, pH = 3 is the best initial pH for the degradation of ENR, PEF and SQX with the degradation efficiencies at 100%, 79.1% and 100% after 180 min, respectively. Considering the degradation rate and electrical energy per order of the test compounds, the UV/H2O2/Fe(II) process was better than the UV/H2O2 and UV/Fe(II) processes because of the greater OH generation. Based on major transformation products of ENR, PEF, and SQX detected during UV/H2O2/Fe(II) treatment, the probable degradation pathway of each compound is proposed. The fluorine atom of ENR and PEF was transformed into fluorine ion, and the sulfur atom was transformed into SO2/SO42-. The nitrogen atom was mainly transformed into NH3/NH4+. Formic acid, acetic acid, oxalic acid, and fumaric acid were identified in the irradiated solutions and all the test compounds and their intermediates can be finally mineralized. In addition, after the UV/H2O2/Fe(II) process, the acute toxicity of the final reaction solutions on zebrafish embryos was lower than that of the initial solution without any treatment. In summary, UV/H2O2/Fe(II) is a safe and efficient technology for antibiotic degradation.

Multicomponent kinetic spectrophotometric determination of pefloxacin and norfloxacin in pharmaceutical preparations and human plasma samples with the aid of chemometrics.

A spectrophotometric method for the simultaneous determination of the important pharmaceuticals, pefloxacin and its structurally similar metabolite, norfloxacin, is described for the first time. The analysis is based on the monitoring of a kinetic spectrophotometric reaction of the two analytes with potassium permanganate as the oxidant. The measurement of the reaction process followed the absorbance decrease of potassium permanganate at 526 nm, and the accompanying increase of the product, potassium manganate, at 608 nm. It was essential to use multivariate calibrations to overcome severe spectral overlaps and similarities in reaction kinetics. Calibration curves for the individual analytes showed linear relationships over the concentration ranges of 1.0-11.5 mg L(-1) at 526 and 608 nm for pefloxacin, and 0.15-1.8 mg L(-1) at 526 and 608 nm for norfloxacin. Various multivariate calibration models were applied, at the two analytical wavelengths, for the simultaneous prediction of the two analytes including classical least squares (CLS), principal component regression (PCR), partial least squares (PLS), radial basis function-artificial neural network (RBF-ANN) and principal component-radial basis function-artificial neural network (PC-RBF-ANN). PLS and PC-RBF-ANN calibrations with the data collected at 526 nm, were the preferred methods--%RPE(T) approximately 5, and LODs for pefloxacin and norfloxacin of 0.36 and 0.06 mg L(-1), respectively. Then, the proposed method was applied successfully for the simultaneous determination of pefloxacin and norfloxacin present in pharmaceutical and human plasma samples. The results compared well with those from the alternative analysis by HPLC.

Identification by virtual screening and functional characterisation of novel positive and negative allosteric modulators of the α7 nicotinic acetylcholine receptor.

Several previous studies have demonstrated that the activity of neurotransmitters acting on ligand-gated ion channels such as the nicotinic acetylcholine receptor (nAChR) can be altered by compounds binding to allosteric modulatory sites. In the case of α7 nAChRs, both positive and negative allosteric modulators (PAMs and NAMs) have been identified and have attracted considerable interest. A recent study, employing revised structural models of the transmembrane domain of the α7 nAChR in closed and open conformations, has provided support for an inter-subunit transmembrane allosteric binding site (Newcombe et al 2017). In the present study, we have performed virtual screening of the DrugBank database using pharmacophore queries that were based on the predicted binding mode of PAMs to α7 nAChR structural models. A total of 81 compounds were identified in the DrugBank database, of which the 25 highest-ranked hits corresponded to one of four previously-identified therapeutic compound groups (carbonic anhydrase inhibitors, cyclin-dependent kinase inhibitors, diuretics targeting the Na+-K+-Cl- cotransporter, and fluoroquinolone antibiotics targeting DNA gyrase). The top-ranked compound from each of these four groups (DB04763, DB08122, furosemide and pefloxacin, respectively) was tested for its effects on human α7 nAChR expressed in Xenopus oocytes using two-electrode voltage-clamp electrophysiology. These studies, conducted with wild-type, mutant and chimeric receptors, resulted in all four compounds exerting allosteric modulatory effects. While DB04763, DB08122 and pefloxacin were antagonists, furosemide potentiated ACh responses. Our findings, supported by docking studies, are consistent with these compounds acting as PAMs and NAMs of the α7 nAChR via interaction with a transmembrane site.

Under ambient UVA exposure, pefloxacin exhibits both immunomodulatory and genotoxic effects via multiple mechanisms.

Pefloxacin (PFLX) is an antibiotic, which shows broad spectrum antimicrobial activities. It is an important derivative of fluoroquinolones (FLQs) group. Ultraviolet radiation (200-400nm) causes major problem for living being which comes at the earth surface naturally through sunlight and increasing regularly due to ozone depletion. PFLX was photodegraded in 5h and forms photoproduct under UVA exposure. At the non photocytotoxic dose PFLX, shows reduced phagocytosis activity, NO (nitric oxide) production, large vacuole formation and down regulated IL-6, TNF-α and IL-1 in BALB/c macrophages at both genes and proteins levels. At higher doses (photocytotoxic doses), PFLX induced a concentration dependent decrease in cell viability of human keratinocyte cell line (HaCaT) and peritoneal macrophages of BALB/c mice. Our molecular docking suggests that PFLX binds only to the cleaved DNA in the DNA-human TOP2A complex. Topoisomerase assay confirmed that PFLX inhibits human topoisomerase by forming an adduct with DNA. Photosensitized PFLX also caused intracellular ROS mediated DNA damage and formation of micronuclei and cyclobutane pyrimidine dimers (CPDs). Increase intracellular ROS leads to apoptosis which was proved through lysosomal destabilization and reduced mitochondrial membrane potential (MMP). Our present study shows that ambient UVA exposure in the presence of PFLX caused immunomodulatory as well as photogenotoxic effects. Therefore, patients under PFLX drug treatment should avoid sunlight exposure, especially during peak hours for their photosafety.

Spectrophotometric determination of pefloxacin mesylate in pharmaceuticals.

A spectrophotometric method is described for assay of pefloxacin mesylate (PFM) in bulk drug and in tablets. The method is based on back extraction of the bromophenol blue dye at pH 5.2 from the dye-drug ion pair followed by measurement of the dye absorbance at 590 nm. The working conditions of the method were investigated and optimized. Beer's law plot showed a good correlation in the concentration range of 0.15-1.25 microg mL(-1). Sensitivity indices such as molar absorptivity, limits of detection and quantification are reported. Intra-day and inter-day precision, and accuracy of the methods were established according to the ICH guidelines, and the er values were in the range of -1.7 to 1.8% with RSD values ranging from 1.0 to 1.1%. The method was successfully applied to the assay of PFM in tablet preparations with recoveries varying from 97.5 to 101.9%, with standard deviation in the range of 0.6 to 1.9. The results were statistically compared with those of the reference method by applying Student's t-test and F-test. Accuracy evaluated by means of the spike recovery method, range from 97.0 to 106.0%, with precision better than 3%.

Ion-activated, Gelrite-based in situ ophthalmic gels of pefloxacin mesylate: comparison with conventional eye drops.

The purpose of our work was to develop an ophthalmic delivery system of a flouroquinolone antibiotic, pefloxacin mesylate, based on the concept of ion-activated in situ gelation. Gelrite gellan gum, a novel ophthalmic vehicle, that gels in the presence of mono- or divalent-cations present in the lacrimal fluid, was used as the gelling agent. The developed formulation was compared with marketed eye drops in efficacy of treatment of bacterial conjunctivitis that was induced artificially in rabbits. The formulations were evaluated for rheological characteristics, in vitro release behavior, antimicrobial efficacy, and efficacy against bacterial conjunctivitis. We found that in situ gelling formulations passed the test for sterility. The formulations exhibited a first-order release pattern over 12 hr in in vitro release studies. The developed formulation was effective against selected micro-organisms in antimicrobial efficacy studies. The shelf lives of formulation was >2 years. The formulation demonstrated better therapeutic efficacy compared with standard eye drops because it improved the clinical parameters monitored for prolonged periods. The developed formulations can be considered as a viable alternative to conventional eye drops.

Preparation and characterization of inclusion complexes of pefloxacin mesylate with three kinds of cyclodextrins.

The ability of alpha-cyclodextrin, beta-cyclodextrin and hydroxypropyl-beta-cyclodextrin (alpha-CD, beta-CD and HP-beta-CD) to break pefloxacin mesylate (PM) aggregates by forming inclusion complexes has been studied using 1H NMR (nuclear magnetic resonance spectroscopy), 13C NMR and fluorescence spectra. The inclusion constants are determined to compare the corresponding inclusion capacity. Solid-inclusion complexes of PM with CDs are synthesized by coprecipitation method, and all the inclusion ratios are found to be 1:1. Additionally, spatial characterization of complexes has been proposed based on two-dimensional nuclear magnetic resonance technique (2D NMR) and spatial conformation is also investigated to propose two possible models between PM and CDs.

Binding interactions of pefloxacin mesylate with bovine lactoferrin and human serum albumin.

The binding of pefloxacin mesylate (PFLX) to bovine lactoferrin (BLf) and human serum albumin (HSA) in dilute aqueous solution was studied using fluorescence spectra and absorbance spectra. The binding constant K and the binding sites n were obtained by fluorescence quenching method. The binding distance r and energy-transfer efficiency E between pefloxacin mesylate and bovine lactoferrin as well as human serum albumin were also obtained according to the mechanism of Förster-type dipole-dipole nonradiative energy-transfer. The effects of pefloxacin mesylate on the conformations of bovine lactoferrin and human serum albumin were also analyzed using synchronous fluorescence spectroscopy.