RESUMO
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.
Assuntos
Antibacterianos , Európio , Estruturas Metalorgânicas , Európio/química , Antibacterianos/análise , Antibacterianos/química , Estruturas Metalorgânicas/química , Quelantes/química , Espectrometria de Fluorescência/métodos , Pefloxacina/análise , Pefloxacina/química , Corantes Fluorescentes/química , Animais , Fluorescência , Resíduos de Drogas/análise , Limite de Detecção , Contaminação de Alimentos/análiseRESUMO
The synthetic antibiotics fluoroquinolones are popular due to their good antibacterial performance and low price, but the risk to human health caused by their residues has attracted great attention. In this study, an ultra-sensitive mAb, 4D7, was prepared with an IC50 of 0.027 ng mL-1 to norfloxacin (NOR) and cross-reactivity of 19.7-47.7% to lomefloxacin (LOM), pefloxacin (PEF), ofloxacin (OFL), enrofloxacin (ENR), ciprofloxacin (CIP), and danofloxacin (DAN). Based on mAb 4D7 and Eu-fluorescent microspheres, a rapid and sensitive immunochromatographic strip was developed for the detection of fluoroquinolone residues in fish and milk. The detection ranges (IC20-IC80) of the strip for the detection of NOR, PEF, LOM, OFL, ENR, CIP and DAN were 0.19-1.1 µg kg-1, 0.39-2.1 µg kg-1, 0.5-2.6 µg kg-1, 0.43-3.3 µg kg-1, 0.61-3.5 µg kg-1, 0.69-5.5 µg kg-1, 0.52-3.4 µg kg-1 in fish, and 0.027-0.19 µg kg-1, 0.049-0.34 µg kg-1, 0.069-0.39 µg kg-1, 0.06-0.41 µg kg-1, 0.089-0.65 µg kg-1, 0.12-0.81 µg kg-1, 0.091-0.52 µg kg-1 in milk, respectively. The recovery rates in spiked sample tests were 88.6-113.6% with a coefficient of variation less than 8.4%. Thus the newly-developed strip was sensitive and reliable for rapid on-site detection of fluoroquinolone residues in fish and milk.
Assuntos
Fluoroquinolonas , Leite , Animais , Humanos , Leite/química , Fluoroquinolonas/análise , Antibacterianos/análise , Pefloxacina/análise , Ciprofloxacina/análise , Norfloxacino , OfloxacinoRESUMO
Intensive use of antibiotics affects biogeochemical cycles and stimulates the evolution of antibiotic resistance, thus threatening global health and social development. The spatiotemporal distributions of antibiotics in single aqueous matrices have been widely documented; however, their occurrence in surface-groundwater systems has received less attention, especially in arid regions that usually have fragile ecosystems. Therefore, we investigated the occurrence of thirty-one antibiotics in the surface water and adjacent groundwater in the Xinjiang Uygur Autonomous Region, China. The results showed that the total concentrations of detected antibiotics varied from 17.37 to 84.09 ng L-1 and from 16.38 to 277.41 ng L-1 in surface and groundwater, respectively. The median concentration of antibiotics showed the pattern of norfloxacin (4.86 ng L-1) > ciprofloxacin (3.93 ng L-1) > pefloxacin (3.39 ng L-1) in surface water; whereas in groundwater, this was in the order of pefloxacin (6.30 ng L-1) > norfloxacin (4.33 ng L-1) > ciprofloxacin (2.68 ng L-1). Heatmap analysis indicated that vertical infiltration had limited effects on antibiotic exchange in surface-ground water systems because of the high potential evaporation and low water storage. Redundancy analysis suggested that the oxidation-reduction potential (p < 0.01) and dissolved oxygen (p < 0.05) jointly affected the distribution of antibiotics in surface water. Ecological risk assessment showed that antibiotics in 98.9% of surface water and 99.1% of groundwater did not pose significant risks to aquatic species. The findings of this study will help develop effective mitigation strategies for antibiotics in aquatic environments.
Assuntos
Água Subterrânea , Poluentes Químicos da Água , Antibacterianos/análise , Norfloxacino , Monitoramento Ambiental , Ecossistema , Poluentes Químicos da Água/análise , Pefloxacina/análise , Ciprofloxacina/análise , Medição de Risco , Água/análise , Oxigênio/análise , ChinaRESUMO
For investigating the spatial, temporal variations and assessing ecological risk of 10 antibiotics and 6 antimycotics, influent sewage water and treated effluent were collected during three different seasons in 19 waste water treatment plants of Tianjin. High performance liquid chromatography tandem mass spectrometry was used to analyze 16 substances. The concentration range of influent samples was not detected (nd) -547.94 ng/L and the concentration range of effluent samples was nd-52.97 ng/L. By calculating the removal efficiency, it was found that Ciprofloxacin (CIP), Ofloxacin (OFL) and Clotrimazole (CTR) were effectively removed. There were significant spatial and temporal differences, the concentration in the dry season was evidently higher than that in the wet and normal seasons, and the northeast was lower than that in the northwest and southeast. By establishing a data set of influent and effluent, the priority features were extracted by feature engineering, which were temperature and NH3-N. Under the condition of ensuring the best performance of the models, the influent model with 9 features and the effluent model with 4 features were established, and the quantitative relationship between the above features and concentration was obtained through partial dependence analysis. Except for Moxifloxacin (MOX), Norfloxacin (NOR) and OFL in the influent samples, the RQ values for other antibiotics and antimycotics were less than 0.1. Among the effluent samples, only NOR had an RQ value greater than 0.1, and OFL, MOX, and Pefloxacin (PEF) had RQ values between 0.01 and 0.1. Comparing the observations and predictions individual RQ values, the predictions were ideal and matched the observations. This work effectively assessed environmental impact and provided a valuable reference for evaluating antibiotics and antimycotics ecological toxicity.
Assuntos
Poluentes Químicos da Água , Purificação da Água , Antibacterianos/análise , Ciprofloxacina , Clotrimazol/análise , Monitoramento Ambiental , Moxifloxacina/análise , Norfloxacino , Ofloxacino/análise , Pefloxacina/análise , Medição de Risco , Esgotos/química , Eliminação de Resíduos Líquidos , Águas Residuárias/química , Poluentes Químicos da Água/análiseRESUMO
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 signiï¬cantly against pefloxacin as predicted. These may provide useful insights for studying antigen-antibody interaction mechanisms to improve antibody affinity maturation in vitro.
Assuntos
Antibacterianos/análise , Anticorpos Monoclonais/química , Ensaio de Imunoadsorção Enzimática/métodos , Região Variável de Imunoglobulina/química , Pefloxacina/análise , Animais , Antibacterianos/química , Antibacterianos/imunologia , Anticorpos Monoclonais/biossíntese , Anticorpos Monoclonais/isolamento & purificação , Afinidade de Anticorpos , Especificidade de Anticorpos , Sítios de Ligação , Ligação Competitiva , Clonagem Molecular , Escherichia coli/genética , Escherichia coli/metabolismo , Expressão Gênica , Hibridomas/química , Hibridomas/imunologia , Imunização , Imunoconjugados/administração & dosagem , Imunoconjugados/química , Região Variável de Imunoglobulina/biossíntese , Região Variável de Imunoglobulina/isolamento & purificação , Limite de Detecção , Camundongos , Camundongos Endogâmicos BALB C , Simulação de Acoplamento Molecular , Mutação , Pefloxacina/química , Pefloxacina/imunologia , Ligação Proteica , Estrutura Secundária de Proteína , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/química , Proteínas Recombinantes/isolamento & purificação , Soroalbumina Bovina/química , Soroalbumina Bovina/imunologia , Homologia Estrutural de ProteínaRESUMO
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.
Assuntos
Galinhas/metabolismo , Cromatografia Líquida de Alta Pressão/métodos , Pefloxacina/análise , Pefloxacina/farmacocinética , Animais , Feminino , Modelos Lineares , Masculino , Pefloxacina/química , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Distribuição TecidualRESUMO
A new chemiluminescence (CL) detection system combined with flow injection analysis (FIA) for the determination of Pefloxacin is proposed. The determination is based on an energy transfer from Pefloxacin to terbium (III). The metal ion enhances the weak CL signal produced by the KMnO4/H2SO3/Pefloxacin system. A modified simplex method was used to optimize chemical and instrumental variables. The influence of the interaction of the permanganate, Tb (III), sodium sulphite and sulphuric acid concentrations, flow rate and injected sample volume was thoroughly investigated by using a modified simplex optimization procedure. The results revealed a strong direct relationship between flow rate and CL intensity throughout the studied range that was confirmed by a gamma test. The response factor for the CL emission intensity was used to assess performance in order to identify the optimum conditions for maximization of the response. Under such conditions, the CL response was proportional to the Pefloxacin concentration over a wide range. The detection limit as calculated according to Clayton's criterion 13.7µgL-1. The analyte was successfully determined in milk samples with an average recovery of 100.6±9.8%.
Assuntos
Análise de Injeção de Fluxo/métodos , Medições Luminescentes/métodos , Leite/metabolismo , Pefloxacina/análise , Permanganato de Potássio/química , Sulfitos/química , Térbio/química , Animais , Bovinos , Leite/químicaRESUMO
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.
Assuntos
Antibacterianos/farmacocinética , Radioisótopos de Carbono/análise , Espectroscopia de Ressonância Magnética/métodos , Pefloxacina/farmacocinética , Administração Oral , Animais , Antibacterianos/análise , Antibacterianos/química , Bile/química , Radioisótopos de Carbono/química , Cromatografia Líquida , Fezes/química , Flúor/química , Masculino , Pefloxacina/análise , Pefloxacina/química , Ratos , Ratos Sprague-Dawley , Espectrometria de Massas em TandemRESUMO
The novel superparamagnetic surface molecularly imprinted Fe3O4@MIP nanoparticles for water-soluble pefloxacin mesylate (PEF-M) were prepared via surface initiated atom transfer radical polymerization (si-ATRP). The binary mixture of methanol and water was selected as the polar solvents for fabricating PEF-M imprinted MIPs. The Fe3O4@MIP exhibited high saturation magnetization of 41.4 emu/g leading to the fast separation. The adsorption behaviors indicated that the Fe3O4@MIP nanoparticles possessed specific recognition and high affinity towards template PEF-M in aqueous media. Moreover, Fe3O4@MIP nanoparticles were directly used to selectively enrich PEF-M from egg samples. By RP-HPLC analysis, the recoveries of PEF-M were obtained as 92.8-96.5% with relative standard division of 2.4-4.0%.
Assuntos
Resíduos de Drogas/isolamento & purificação , Ovos/análise , Nanopartículas de Magnetita/química , Impressão Molecular/métodos , Pefloxacina/isolamento & purificação , Ácido Acético/química , Adsorção , Animais , Galinhas , Resíduos de Drogas/análise , Resíduos de Drogas/química , Resíduos de Drogas/metabolismo , Nanopartículas de Magnetita/ultraestrutura , Metanol/química , Pefloxacina/análise , Pefloxacina/química , Pefloxacina/metabolismo , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectroscopia de Infravermelho com Transformada de Fourier , Propriedades de SuperfícieRESUMO
Pefloxacin mesylate, a broad-spectrum antibacterial fluoroquinolone, has been widely used in clinical practice. Therefore, it is very important to detect the concentration of Pefloxacin mesylate. In this research, the near-infrared spectroscopy (NIRS) has been applied to quantitatively analyze on 108 injection samples, which was divided into a calibration set containing 89 samples and a prediction set containing 19 samples randomly. In order to get a satisfying result, partial least square (PLS) regression and principal components regression (PCR) have been utilized to establish quantitative models. Also, the process of establishing the models, parameters of the models, and prediction results were discussed in detail. In the PLS regression, the values of the coefficient of determination (R(2)) and root mean square error of cross-validation (RMSECV) of PLS regression are 0.9263 and 0.00119, respectively. For comparison, though applying PCR method to get the values of R(2) and RMSECV we obtained are 0.9685 and 0.00108, respectively. And the values of the standard error of prediction set (SEP) of PLS and PCR models are 0.001480 and 0.001140. The result of the prediction set suggests that these two quantitative analysis models have excellent generalization ability and prediction precision. However, for this PFLX injection samples, the PCR quantitative analysis model achieved more accurate results than the PLS model. The experimental results showed that NIRS together with PCR method provide rapid and accurate quantitative analysis of PFLX injection samples. Moreover, this study supplied technical support for the further analysis of other injection samples in pharmaceuticals.
Assuntos
Pefloxacina/análise , Preparações Farmacêuticas/química , Análise de Componente Principal , Espectroscopia de Luz Próxima ao Infravermelho/métodos , Calibragem , Análise dos Mínimos Quadrados , Modelos Químicos , Análise MultivariadaRESUMO
A novel chemiluminescence (CL) reaction system with bis(hydrogenperiodato) argentate(III) complex anion (Ag(III) complex, [Ag(HIO(6))(2)](5-)), for the first time, is developed for the determination of lomefloxacin (LMFX), enrofloxacin (ENLX) and pefloxacin (PFLX). The possible CL emission mechanism was discussed by comparing the fluorescence emission with CL spectra. The CL conditions of [Ag(HIO(6))(2)](5-)-H(2)SO(4)-LMFX/ENLX/PFLX systems were investigated and optimized. Under the optimized experimental conditions, the CL intensity is proportional to the concentration of the drugs in the range 0.2994-36.80x10(-7)g mL(-1) for LMFX, 4.00-30.0x10(-7)g mL(-1) for ENLX and 1.54-27.64x10(-7)g mL(-1) for PFLX. The limit of detection (s/n=3) was 9.1x10(-9)g mL(-1) for LMFX, 3.1x10(-9)g mL(-1) for ENLX and 4.4x10(-9)g mL(-1) for PFLX. The recovery of LMFX, ENLX and PELX from the spiked pharmaceutical preparations was in the range of 92.3-105% with the RSDs of 0.5-2.7%. For urine, serum and milk samples the recoveries of the three drugs were in the range of 85.1-107% for LMFX with the RSDs of 2.3-3.4%. 80.2-112% for ENLX with the RSDs of 1.4-2.8%, and 87.8-114% for PFLX with the RSDs of 1.6-2.7%. The proposed method was applied successfully to the determination of these compounds in real samples.
Assuntos
Fluoroquinolonas/análise , Luminescência , Compostos Organometálicos/química , Antibacterianos/análise , Enrofloxacina , Métodos , Pefloxacina/análise , Ácido Periódico/química , Prata/químicaRESUMO
The anodic behavior and determination of pefloxacin on boron-doped diamond and glassy carbon electrodes were investigated using cyclic, linear sweep, differential pulse and square wave voltammetric techniques. In cyclic voltammetry, pefloxacin shows one main irreversible oxidation peak and additional one irreversible ill-defined wave depending on pH values for both electrodes. The results indicate that the process of pefloxacin is irreversible and diffusion controlled on boron-doped diamond electrode and irreversible but adsorption controlled on glassy carbon electrode. The peak current is found to be linear over the range of concentration 2x10(-6) to 2x10(-4)M in 0.5M H(2)SO(4) at about +1.20V (versus Ag/AgCl) for differential pulse and square wave voltammetric technique using boron-doped diamond electrode. The repeatability, reproducibility, precision and accuracy of the methods in all media were investigated. Selectivity, precision and accuracy of the developed methods were also checked by recovery studies. The procedures were successfully applied to the determination of the drug in pharmaceutical dosage forms and humans serum samples with good recovery results. No electroactive interferences from the excipients and endogenous substances were found in the pharmaceutical dosage forms and biological samples, respectively.
Assuntos
Eletroquímica/métodos , Pefloxacina/análise , Carbono , Diamante , Eletrodos , Humanos , Pefloxacina/sangue , Preparações Farmacêuticas/química , Reprodutibilidade dos TestesRESUMO
An efficient method was developed for simultaneous determination of 13 quinolones--namely, enoaxacin (ENO), marbofloxacin (MAR), ciprofloxacin (CIP), norfloxacin (NOR), ofloxacin (OFL), pefloxacin methanesulfonate (PEF), danofloxacin (DAN), enrofloxacin (ENR), lomefloxacin (LOM), difloxacin (DIF), sarafloxacin (SAR), oxolinic acid (OXO), and flumequine (FLU)--in eggs by column liquid chromatography/electrospray ionization-tandem mass spectrometry. Samples were extracted with a phosphoric acid-phosphate buffer followed by purification with a solid-phase extraction cartridge. Recoveries for the 13 quinolones were 67-93% with intraday and interday coefficients of variation ranging from 4 to 9% and 2 to 18%, respectively. The limit of determination was 0.05 microg/kg for OXO and FLU; 0.1 microg/kg for MAR, OFL, CIP, LOM, DAN, SAR, DIF, NOR, and ENR; and 0.2 microg/kg for ENO and PEF. The method was also applied to study the depletion of PEF in eggs. The concentration of PEF increased and reached a maximum value on the third day, and then decreased rapidly until it could not be detected on day 32; its metabolite NOR was detectable on the second day, and then reached a maximum on the sixth day, after which it could not be detected until day 15.
Assuntos
Antibacterianos/análise , Ovos/análise , Pefloxacina/análise , Quinolonas/análise , Animais , Cromatografia Líquida de Alta Pressão , Feminino , Concentração de Íons de Hidrogênio , Indicadores e Reagentes , Padrões de Referência , Reprodutibilidade dos Testes , Soluções , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em TandemRESUMO
A confirmative method was developed with high performance liquid chromatography coupled with electrospray ionization tandem mass spectrometry (HPLC-MS/MS) to simultaneously detect 16 quinolone residues in animal tissues, which included nalidixinic acid, oxolinic acid, flumequine, norfloxacin, enoxacin, ciprofloxacin, lomefloxacin, danofloxacin, enrofloxacin, ofloxacin, sarafloxacin, difloxacin, marbofloxacin, pefloxacin, sparfloxacin and orbifloxacin. In the method, the 16 residues were extracted with acidified acetonitrile, cleaned-up with hexane, and concentrated with a rotary evaporator. Then the reconstituted sample solution was analyzed using HPLC-MS/MS in positive mode, with a Inertsil C8-3 column as the analytical column. The method was validated at 10, 50 and 100 microg/kg. The validation results were as follows: the linear ranges were from 10 to 100 microg/kg; the overall recoveries were from 62.4% to 102% with the relative standard deviations of 1.4%-11.9%. The method is simple, rapid, and accurate, and its performance could meet the requirements of the domestic and international legislation. The method is applicable to simultaneously confirm multi-residues of quinolones in animal tissues such as chicken muscle, chicken liver and fish muscle.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Quinolonas/análise , Espectrometria de Massas por Ionização por Electrospray/métodos , Animais , Ciprofloxacina/análogos & derivados , Ciprofloxacina/análise , Enoxacino/análise , Enrofloxacina , Fluoroquinolonas/análise , Norfloxacino/análise , Ofloxacino/análise , Pefloxacina/análise , Reprodutibilidade dos TestesRESUMO
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%.
Assuntos
Anti-Infecciosos/análise , Pefloxacina/análise , Espectrofotometria/métodos , Algoritmos , Anti-Infecciosos/química , Azul de Bromofenol/química , Clorofórmio/química , Concentração de Íons de Hidrogênio , Estrutura Molecular , Pefloxacina/química , Reprodutibilidade dos Testes , Solubilidade , Solventes/químicaRESUMO
Pefloxacin has been increasingly used in veterinary medicine to treat microbial infections. To avoid using a labor-intensive instrumental method to detect the residue of pefloxacin in food, a simple and convenient indirect competitive enzyme-linked immunosorbent assay method has been developed in this study. The antibody generated from immunogen cationized bovine serum albumin-pefloxacin showed high sensitivity toward pefloxacin with an IC50 value of 6.7 ppb in buffer and was suitable for a screening assay to detect the residue of pefloxacin in food products. The antibody has been assessed using rapid enzyme immunoassays to exploit its specificity. The antibody prepared shows cross-reactivity with a few other (fluoro)quinolones including fleroxacin (116%), enrofloxacin (88%), and ofloxacin (10%). The assay measured drug residue in chicken liver spiked with pefloxacin with an interassay coefficient of variation of 13.6% or less and an intra-assay coefficient of variation of 10.9% or less. The average recovery rates at 0.5, 5, 10, 50, and 100 ppb were in the range of 86-106% for interassay and in the range of 87-103% for intra-assay, respectively.
Assuntos
Anticorpos/imunologia , Galinhas , Ensaio de Imunoadsorção Enzimática/métodos , Fígado/química , Pefloxacina/análise , Pefloxacina/imunologia , Animais , Especificidade de Anticorpos , Ligação Competitiva , Resíduos de Drogas/análise , Masculino , Coelhos , Sensibilidade e EspecificidadeRESUMO
The objective of this research was to develop and validate analytical methods for quantitative determination of fluoroquinolones of third generation. Simple and rapid chromatographic method was developed and validated for quantitative determination of four quinolone antibiotics in tablets and injection preparations. The fluoroquinolones studied were gatifloxacin (GAT), levofloxacin (LEV), lomefloxacin (LOM) and pefloxacin (PEF). The quinolones were analyzed by using a LiChrospher 100 RP-18 column (5 microm, 125 mm x 4 mm) and a mobile phase constituted of water:acetonitrile (80:20, v/v) with 0.3% of triethylamine and pH adjusted to 3.3 with phosphoric acid. The flow rate was 1.0 mL/min and the analyses were performed using UV detector with wavelengths varying from 279 to 295 nm. The analyses were performed at room temperature (24 +/- 2 degrees C). All fluoroquinolones were separated within 5 min. The calibration curves were linear (r>or=0.9999) over a concentration range from 4.0 to 24.0 microg/mL. The relative standard deviation (R.S.D.) was < 1.0% and average recovery was above 99.54%.
Assuntos
Antibacterianos/análise , Química Farmacêutica/métodos , Cromatografia Líquida de Alta Pressão/métodos , Fluoroquinolonas/análise , Levofloxacino , Ofloxacino/análise , Pefloxacina/análise , Quinolonas/análise , Calibragem , Técnicas de Química Analítica/métodos , Cromatografia , Etilaminas/análise , Gatifloxacina , Concentração de Íons de Hidrogênio , Modelos Químicos , Modelos Estatísticos , Preparações Farmacêuticas/análise , Análise de Regressão , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrofotometria Ultravioleta/métodos , Comprimidos , Temperatura , Fatores de TempoRESUMO
A liquid chromatographic method with fluorescence detection was developed for simultaneous determination of norfloxacin, ofloxacin, ciprofloxacin, pefloxacin, lomefloxacin, danofloxacin, enrofloxacin, sarafloxacin, difloxacin, oxolinic acid, and flumequine in milk The samples were extracted with 10% trichloroacetic acid/acetonitrile (9 + 1, v/v) and cleaned by Strata-X reversed-phase solid-phase extraction cartridges. The 11 quinolones were separated on a reversed-phase C18 column (Hypersil BDS-C18) with mobile phase gradient elution and detected with fluorescence by means of a wavelength program. The recoveries for milk fortified with the 11 quinolones at 3 levels were 69-88% with acceptable relative standard deviations of <9% (intraday) and <14% (interday). The limits of detection were 23 microg/L for enrofloxacin, and 1-9 microg/L for the other 10 quinolones.
Assuntos
Cromatografia Líquida/métodos , Leite/química , Quinolonas/análise , Quinolonas/química , Espectrometria de Fluorescência/métodos , Animais , Calibragem , Técnicas de Química Analítica/métodos , Cromatografia , Ciprofloxacina/análogos & derivados , Ciprofloxacina/análise , Enrofloxacina , Fluoroquinolonas/análise , Leite/metabolismo , Norfloxacino/análise , Ofloxacino/análise , Ácido Oxolínico/análise , Pefloxacina/análise , Padrões de Referência , Reprodutibilidade dos Testes , Solventes/química , Fatores de TempoRESUMO
A new highly sensitive fluorescence spectrophotometic method for the determination of pefloxacin (PEFX) has been developed based on micella enhancement. The fluorescence characteristic of micelle inclusion complex formed between PEFX and sodium dodecyl sulfate (SDS) was studied. Different variables and parameters affecting the fluorescence were studied and optimized. Experiments show that sodium dodecyl sulfate can enhance greatly the fluorescence signal for pefloxacin in pH 5 Britton Robinson(BR) buffer solution. The fluorimetric method allows the determination of 0.06-1.20 microg x mL(-1) of pefloxacin with SDS in aqueous solution with lambda ex = 278 nm and lambda em = 432 nm, respectively. The detection limit for PEFX is 0.06 microg x mL(-1), The recoveries are 98.5%-100.8% and the RSDs are 1.4%-2.3%. The proposed procedures could be applied successfully to the determination of the pefloxacin drugs in tablets and human plasma with good recovery. In this paper we determined pefloxacin in human plasma by synchronization-derivative fluorescence spectroscopic techniques with good analytical selectivity, high sensitivity, high capability of eliminating blank interference and improving the limit of detection.
Assuntos
Antibacterianos/química , Fluorescência , Fluorometria/métodos , Pefloxacina/química , Espectrometria de Fluorescência/métodos , Antibacterianos/análise , Química Farmacêutica/métodos , Humanos , Limite de Detecção , Micelas , Pefloxacina/análiseRESUMO
Two simple, quick and sensitive spectrophotometric methods are described for the determination of enrofloxacin and Pefloxacin. The methods are based on the reaction of these drugs with bromophenol blue (BPB) and methyl orange (MO) in buffered aqueous solution at pH 2.3-2.5 in case of bromophenol blue and at pH 3.6 with MO to give highly coloured complex species, extractable with chloroform. The coloured products are quantitated spectrophotometrically at 420 and 424 nm for BPB and MO, respectively. Optimisation of the different experimental conditions is described. Beer's law is obeyed in the concentration ranges 2-12 and 2-18 microg ml(-1) with BPB and in the ranges 1-12 and 4-40 microg ml(-1)with MO for enrofloxacin and pefloxacin, respectively. The proposed methods are applied for determination of Enroxil oral solution, Peflacine tablets and Peflacine ampoules with mean percentage accuracies 99.5+/-0.99, 99.39+/-1.05 and 100.02+/-0.895, respectively, with BPB and 100.30+/-0.89, 100.25+/-0.98 and 100.20+/-0.72, respectively, with MO.