RESUMO
In this study, we proposed a novel method utilizing polyethyleneimine (PEI)-modified halloysite nanotubes (HNTs)-based hybrid silica monolithic spin tip to analyze hydrophilic ß-lactam antibiotics and ß-lactamases inhibitors in whole blood samples for the first time. HNTs were incorporated directly into the hybrid silica monolith via a sol-gel method, which improved the hydrophilicity of the matrix. The as-prepared monolith was further modified with PEI by glutaraldehyde coupling reaction. It was found that the PEI-modified HNTs-based hybrid silica monolith enabled a large adsorption capacity of cefoperazone at 35.7 mg g-1. The monolithic spin tip-based purification method greatly reduced the matrix effect of whole blood samples and had a detection limit as low as 0.1 - 0.2 ng mL-1. In addition, the spiked recoveries of sulbactam, cefuroxime, and cefoperazone in blank whole blood were in the range of 89.3-105.4 % for intra-day and 90.6-103.5 % for inter-day, with low relative standard deviations of 1.3-7.2 % and 4.9-10.5 %, respectively. This study introduces a new strategy for preparing nanoparticles incorporated in a hybrid silica monolith with a high adsorption capacity. Moreover, it offers a valuable tool to monitor sulbactam, cefoperazone, and cefuroxime in whole blood from pregnant women with the final aim of guiding their administration.
Assuntos
Cefoperazona , Cefuroxima , Interações Hidrofóbicas e Hidrofílicas , Limite de Detecção , Nanotubos , Dióxido de Silício , Extração em Fase Sólida , Sulbactam , Cefoperazona/sangue , Cefoperazona/química , Humanos , Sulbactam/sangue , Sulbactam/química , Extração em Fase Sólida/métodos , Dióxido de Silício/química , Nanotubos/química , Cefuroxima/sangue , Cefuroxima/química , Argila/química , Adsorção , Antibacterianos/sangue , Antibacterianos/química , Polietilenoimina/química , Cromatografia Líquida de Alta Pressão/métodos , Reprodutibilidade dos TestesRESUMO
Antimicrobial activity of cefoperazone, a high protein bound cephalosporin, depends on its unbound concentration. However, the protein binding data of cefoperazone in children is limited, making it challenging to optimize antimicrobial therapy in pediatric clinical practice. Furthermore, a validated method to measure the free part in children is unavailable with the small volume of samples that can be obtained. Therefore, in the present study, we developed and validated an LC-MS/MS method for the determination of free cefoperazone in children. In this study, 70⯵L of plasma was used to prepare the ultrafiltrate (only containing the free drug). Chromatographic separation of the analyte was achieved on a C18 column using gradient elution with a mobile phase of acetonitrile and water (0.1% formic acid). Negative electrospray ionisation in the multiple reaction monitoring mode was applied for the detection of cefoperazone and ceftiofur (internal standard). The calibration curve was prepared in the range of 5-5000â¯ng/mL with excellent linearity. For each level of quality control samples, the intra- and inter-day precision (CV) was below 9.0%, and the accuracy ranged from 91.5% to 105.0%. The matrix effect was less than 11.7%, and the recovery was between 92.9% and 95.9% of cefoperazone. The validated method has been successfully applied to the determination of free plasma concentration of cefoperazone in pediatric patients. The results of the unbound fraction showed considerable individual variability (range: 8.1-48.0%). The correlation analysis showed that age and albumin had significant effects on the protein binding of cefoperazone.
Assuntos
Cefoperazona/sangue , Fatores Etários , Técnicas Biossensoriais/métodos , Criança , Pré-Escolar , Cromatografia Líquida de Alta Pressão , Feminino , Humanos , Lactente , Recém-Nascido , Limite de Detecção , Masculino , Ligação Proteica , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Espectrometria de Massas em TandemRESUMO
BACKGROUND: Critically ill patients show several pathophysiological alterations that can complicate antibiotic dosing. Hence, there is a strong rationale to individualize anti-infective dosing in these patients by using therapeutic drug monitoring (TDM). The current study aimed to develop and validate a liquid chromatography-tandem mass spectrometry method for the simultaneous determination of total and unbound plasma concentrations of 3 commonly used antibiotics (meropenem, imipenem/cilastatin, and cefoperazone/sulbactam) in the treatment of infections in critically ill patients in China, which could be suitable for routine TDM in hospital laboratories. METHODS: The unbound drug was separated from the bound drug by ultrafiltration. Simple protein precipitation was used for sample preparation. Meropenem, imipenem/cilastatin, cefoperazone/sulbactam, and their corresponding internal standards were then resolved using the Waters CORTECS C18 column. All the compounds were detected using electrospray ionization in the positive/negative ion-switching mode. RESULTS: The calibration curves were linear for all compounds, with correlation coefficients (R) above 0.99 for total concentrations in human plasma and unbound concentrations in the plasma ultrafiltrate. For both total and unbound drugs, the relative errors and intra-assay/interassay relative standard deviations were below 15%. The limit of quantification was 0.05 mcg/mL for both total plasma concentrations and plasma ultrafiltrate concentrations of all compounds. CONCLUSIONS: The method was simple, rapid, and reliable and is currently being used to provide a TDM service to enhance the efficacious use of the 3 antibiotics.
Assuntos
Cefoperazona/sangue , Combinação Imipenem e Cilastatina/sangue , Cilastatina/sangue , Imipenem/sangue , Meropeném/sangue , Sulbactam/sangue , Cromatografia Líquida de Alta Pressão/métodos , Estado Terminal , Monitoramento de Medicamentos/métodos , Humanos , Plasma/química , Espectrometria de Massas em Tandem/métodosRESUMO
A dual-channel microchip electrophoresis (ME) with in-channel amperometric detection was developed for cefoperazone and sulbactam determination simultaneously. In this study, a microelectrode detector was made of gold nanoparticles (GNPs) modified indium tin oxide (ITO)-coated poly-ethylene terephthalate (PET) film. The parameters including detection potential applied on working electrode, buffer concentration and pH value were optimized to improve the detection sensitivity and separation efficiency of cefoperazone and sulbactam. Under the optimal conditions, sensitive detection of cefoperazone and sulbactam was obtained with limits of detection (LODs) (S/N = 3) of 0.52 and 0.75 µg/mL, respectively. The plasma sample, which was from a patient with a brain injury taking Sulperazone, was successfully detected with a simple sample pretreatment process by dual-channel ME amperometric detection. This rapid and sensitive method possesses practical potential in clinical applications, and could provide a guidance for clinical rational drug use.
Assuntos
Cefoperazona/análise , Eletroforese em Microchip/instrumentação , Sulbactam/análise , Métodos Analíticos de Preparação de Amostras , Soluções Tampão , Cefoperazona/sangue , Cefoperazona/química , Eletroquímica , Humanos , Concentração de Íons de Hidrogênio , Sulbactam/sangue , Sulbactam/química , Fatores de TempoRESUMO
The plasma disposition of cefoperazone was investigated after intravenous (IV) and intramuscular (IM) administrations of 20 mg/kg as a single dose in six camels (Camelus dromedarius) in a crossover design. Blood plasma samples were analysed by high-performance liquid chromatography (HPLC). After IV administration, elimination half-life (t1/2ß), volume of distribution at steady state (Vdss), total body clearance (Cltot) and mean residence time (MRT) of cefoperazone were 1.95 h, 0.38 L/kg, 0.17 L/h/kg and 2.16 h, respectively. After IM administration of cefoperazone, peak plasma concentration (Cmax) was 21.95 µg/mL and it was obtained at (tmax) 1.23 h. Absorption half-life (t1/2ab), elimination half-life and mean absorption time were 0.45 h, 2.84 h and 2.07 h, respectively. The bioavailability of cefoperazone was 89.42%. The lack of local reaction or any other adverse effects and the very good bioavailability following IM administration indicate that cefoperazone might be a promising alternative treatment for a variety of infectious diseases in camels.
Assuntos
Antibacterianos/sangue , Camelus/sangue , Cefoperazona/sangue , Animais , Antibacterianos/farmacocinética , Área Sob a Curva , Disponibilidade Biológica , Cefoperazona/farmacocinética , Estudos Cross-Over , Meia-Vida , Injeções Intramusculares , Injeções Intravenosas , MasculinoRESUMO
A rapid, accurate and specific high-performance liquid chromatography-tandem mass spectrometry method has been validated for the simultaneous determination of cefoperazone and sulbactam in a small volume sample for children. A Shim-pack XR-ODS C18 column with gradient elution of water (0.1% formic acid) and acetonitrile (0.1% formic acid) solution was used for separation at a flow rate of 0.3 mL/min. The calibration curves of two analytes in serum showed excellent linearity over the concentration ranges of 0.03-10 µg/mL for cefoperazone, and 0.01-3 µg/mL for sulbactam, respectively. This method involves simple sample preparation steps and was validated according to standard US Food and Drug Administration and European Medicines Agency guidelines in terms of selectivity, linearity, detection limits, matrix effects, accuracy, precision, recovery and stability. This assay can be easily implemented in clinical practice to determine concentrations of cefoperazone and sulbactam in children.
Assuntos
Cefoperazona/sangue , Cromatografia Líquida de Alta Pressão/métodos , Monitoramento de Medicamentos/métodos , Sulbactam/sangue , Espectrometria de Massas em Tandem/métodos , Cefoperazona/química , Cefoperazona/farmacocinética , Criança , Pré-Escolar , Estabilidade de Medicamentos , Humanos , Limite de Detecção , Modelos Lineares , Reprodutibilidade dos Testes , Sulbactam/química , Sulbactam/farmacocinéticaRESUMO
PURPOSE: Cefoperazone/sulbactam (CFP/SUL) is a ß-lactam/ß-lactamase inhibitor combination with little data available for the development of effective dosing guidelines during continuous renal replacement therapy. This study aimed to investigate the pharmacokinetics (PK) of cefoperazone/sulbactam in critically ill patients on continuous venovenous hemofiltration (CVVH). METHODS: A prospective, single-center, and open-label study was conducted. Critically ill patients receiving CVVH with 3 g cefoperazone/sulbactam (2.0/1.0 g) intravenously every 8 h were recruited. Serial blood and ultrafiltrate samples were paired collected for initial dose (occasion 1) and steady state (occasion 2). PK was assessed by non-compartmental analysis, and pharmacodynamics (PD) was evaluated by the percent of time for which drug concentrations exceed the minimum inhibitory concentration (%T >MIC). RESULTS: Total fourteen patients were enrolled. Volume of distribution at steady state (V ss) of cefoperazone and sulbactam for initial doses (20.8 ± and 28.4 L, respectively) increased significantly compared with those in healthy volunteers (P = 0.009 for CFP, P = 0.030 for SUL). Both cefoperazone and sulbactam showed significantly lower total clearance (CLt) (46.2 and 117.6 mL/min, respectively) compared with healthy volunteers (P = 0.000 for CFP, P = 0.017 for SUL). There is no significant difference in PK between occasion 1 and occasion 2 (P > 0.05). For occasion 1, mean CVVH clearance accounted for 34.3 and 33.9 % for CLt of cefoperazone and sulbactam, respectively. The minimum PD target of 60%T >MIC was achieved in seven of eight patients. For occasion 2, eight of nine patients achieved cefoperazone concentrations that were above the MIC for the entire dosing interval. CONCLUSIONS: PK of cefoperazone/sulbactam was altered in critically ill patients undergoing CVVH. Therapeutic drug monitoring would be recommended to individualize the dose regimen.
Assuntos
Antibacterianos/farmacocinética , Cefoperazona/farmacocinética , Sulbactam/farmacocinética , Acinetobacter baumannii/efeitos dos fármacos , Acinetobacter baumannii/crescimento & desenvolvimento , Adulto , Idoso , Antibacterianos/sangue , Antibacterianos/farmacologia , Cefoperazona/sangue , Cefoperazona/farmacologia , Estado Terminal , Feminino , Hemofiltração , Humanos , Masculino , Testes de Sensibilidade Microbiana , Pessoa de Meia-Idade , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/crescimento & desenvolvimento , Sulbactam/sangue , Sulbactam/farmacologia , Adulto JovemRESUMO
BACKGROUND: To identify changes in cefoperazone/sulbactam penetration into cerebrospinal fluid (CSF) after craniotomy and to investigate preliminarily whether cefoperazone/sulbactam CSF concentration can reach therapeutic level when administered intravenously after neurosurgical operation. METHODS: Neurosurgical patients with an indwelling ventricular drainage pipe who received prophylactic cefoperazone/sulbactam for the treatment of intracranial infection were received a cefoperazone/sulbactam 2:1, 3.0-g infusion for 3 hours every 6 hours for 24 h. Venous blood and CSF specimens were collected to determine cefoperazone/sulbactam concentrations. RESULTS: The cefoperazone and sulbactam concentrations in serum were highest at the second hour (237.54 ± 336.72 mg/L and 66.52 ± 80.38 mg/L, respectively) and then decreased. The cefoperazone and sulbactam concentrations in CSF were highest at the 4th hour (39.22 ± 75.55 mg/L and 6.24 ± 8.35 mg/L, respectively) and then decreased. CSF penetration measured by the ratio of peak concentrations (CSF/serum) was 8.6% ± 7.2% for cefoperazone and 13.5% ± 11.9% for sulbactam, CSF penetration measured by the ratio of trough concentrations (CSF/serum) was 13.4% ± 5.3% for cefoperazone and 106.5% ± 87.5% for sulbactam. CSF penetration represented by the ratio of area under the curve (AUC) of CSF and serum was 14.5% for cefoperazone and 22.6% for sulbactam. Neurosurgical impairment of the blood-brain barrier may improve the CSF penetration of these drugs, but it is difficult to reach the MIC90 of resistant bacteria. If single intravenous administration time was extended to 3 hours, the serum concentrations of drugs were able to meet the PK/PD standard (T> MIC%> 50%) for treating common, highly resistant bacteria. CONCLUSIONS: The CSF penetration of cefoperazone/sulbactam may be enhanced after neurosurgical impairment of the blood-brain barrier. This study is a pilot research of cefoperazone/sulbactam using in neurosurgical individuals, However, it needs to be confirmed by further large-scale studies.
Assuntos
Cefoperazona/sangue , Cefoperazona/líquido cefalorraquidiano , Craniotomia , Sulbactam/sangue , Sulbactam/líquido cefalorraquidiano , Adulto , Idoso , Anti-Infecciosos Urinários/administração & dosagem , Anti-Infecciosos Urinários/sangue , Anti-Infecciosos Urinários/líquido cefalorraquidiano , Cefoperazona/administração & dosagem , Combinação de Medicamentos , Feminino , Humanos , Infusões Intravenosas , Masculino , Pessoa de Meia-Idade , Projetos Piloto , Teste Bactericida do Soro , Sulbactam/administração & dosagem , Fatores de TempoRESUMO
Imipenem, cefaperazon-sulbactam and cefepime are the antibiotics of choice for the treatment of soft tissue infections due to Acinetobacter baumannii. In this study, it was aimed to determine the invivo and invitro efficacy of, these antibiotics against drug susceptible and multidrug resistant A.baumannii in an experimental abscess model. Abscess models were established in Wistar-Albino type female rats. Susceptibility tests were performed by E-test. Rats were divided randomly into four groups with eight rats in one group. Standard absorbent paper discs containing 6 log10 CFU microorganisms were used to form an abscess model. The first group was regarded as the control group and the other three groups were the study group each treated with one of the test antibiotics. Cardiac blood samples for serum antibiotic efficacy test, were obtained on the fourth day of treatment and 30 minutes after the last dose. The number of live bacteria at the area of infection was determined by colony count method. All of the three antibiotics reached sufficient concentration in sera of rats and there were no statistically important difference between the efficacies of these antibiotics (p= 0.778). In all of the antibiotic-treated groups, the weight of the abscess material were less, macroscopic views were smaller and the colony counts per gram of abscess tissue were less than the control group (p< 0.001). All antibiotics were effective against susceptible and resistant strains in vitro. No resistance was detected against imipenem, cefaperazon-sulbactam and cefepime in the course of therapy. Cefaperazone-sulbactam and cefepime were as effective as imipenem against susceptible and multi-drug resistant A.baumannii both in vivo and in vitro. Since irrational use of extended spectrum cephalosporins are frequently associated with the emergence of carbapenem resistant strains, the use of relatively narrow spectrum antibiotics should better be considered in the empirical treatment of A.baumannii infections.
Assuntos
Abscesso/tratamento farmacológico , Infecções por Acinetobacter/tratamento farmacológico , Acinetobacter baumannii/efeitos dos fármacos , Antibacterianos/uso terapêutico , Infecções dos Tecidos Moles/tratamento farmacológico , Abscesso/microbiologia , Infecções por Acinetobacter/microbiologia , Animais , Antibacterianos/sangue , Antibacterianos/farmacologia , Cefepima , Cefoperazona/sangue , Cefoperazona/farmacologia , Cefoperazona/uso terapêutico , Cefalosporinas/sangue , Cefalosporinas/farmacologia , Cefalosporinas/uso terapêutico , Contagem de Colônia Microbiana , Modelos Animais de Doenças , Combinação de Medicamentos , Farmacorresistência Bacteriana Múltipla , Feminino , Imipenem/sangue , Imipenem/farmacologia , Imipenem/uso terapêutico , Distribuição Aleatória , Ratos , Ratos Wistar , Teste Bactericida do Soro , Infecções dos Tecidos Moles/microbiologia , Sulbactam/sangue , Sulbactam/farmacologia , Sulbactam/uso terapêutico , Coxa da PernaRESUMO
A rapid and highly sensitive liquid chromatography-tandem mass spectrometric (LC-MS/MS) method for simultaneous determination of cefoperazone sodium and sulbactam sodium in human plasma was developed. The analytes and internal standard (IS), cefuroxime sodium, were extracted from human plasma via liquid-liquid extraction with ethyl acetate and separated on a Waters Xterra C18 column within 3.5 min. Quantitation was performed on a triple quadrupole mass spectrometer employing electrospray ionization technique, operating in selected reaction monitoring (SRM) and negative ion mode. The precursor to product ion transitions monitored for cefoperazone, sulbactam and IS were m/z 644.1â528.0, 232.1â140.0, and 423.0â362.0, respectively. The assay was validated in the linear range of 0.1-20 µg/mL for cefoperazone and 0.02-4 µg/mL for sulbactam. The intra- and inter-day precisions (CV%) were within 8.39% for each analyte. The recoveries were greater than 87.3% for cefoperazone and 87.2% for sulbactam. Each analyte was found to be stable during all sample storage, preparation and analytical procedures. The method was successfully applied in a pharmacokinetic study of Sulperazon injection in six hospital-acquired pneumonia (HAP) patients.
Assuntos
Antibacterianos/sangue , Cefoperazona/sangue , Cromatografia Líquida de Alta Pressão/métodos , Sulbactam/sangue , Espectrometria de Massas em Tandem/métodos , Antibacterianos/farmacocinética , Cefoperazona/farmacocinética , Combinação de Medicamentos , Humanos , Espectrometria de Massas por Ionização por Electrospray/métodos , Sulbactam/farmacocinéticaRESUMO
Wide-spectrum quinolones such as moxifloxacin and levofloxacin as well as high-order cephalosporins such as cefoperazone and cefepime have increased antimicrobial activity. However, little is known about their distribution in fetal blood. Therefore, the aim of this study was to measure and compare maternal and fetal blood levels of these agents. For the measurement of blood levels, 9 pregnant women received cefepime hydrochloride, 10 received cefoperazone, 10 received moxifloxacin and 12 received levofloxacin intravenously. Maternal and umbilical cord blood samples were drawn during delivery. Antibiotic levels were analysed by high-performance liquid chromatography. Mean transplacental passage rates of moxifloxacin, levofloxacin, cefepime and cefoperazone were 74.84%, 66.53%, 23.21% and 12.68%, respectively, and mean transfetal passage rates were 90.78%, 84.22%, 79.17% and 79.78%, respectively. The transplacental passage rate for either quinolone was significantly higher than that of either cephalosporin, and the transplacental passage rate of cefoperazone was lower than that of cefepime. In conclusion, both quinolones have high transplacental passage rates. Cefepime and cefoperazone have a lower transplacental passage rate and thus may be used as prophylaxis in situations where transplacental passage is undesirable.
Assuntos
Antibacterianos/sangue , Compostos Aza/sangue , Cefoperazona/sangue , Sangue Fetal/química , Levofloxacino , Troca Materno-Fetal , Ofloxacino/sangue , Quinolinas/sangue , Adulto , Antibacterianos/administração & dosagem , Antibacterianos/farmacocinética , Compostos Aza/administração & dosagem , Compostos Aza/farmacocinética , Cefepima , Cefoperazona/administração & dosagem , Cefoperazona/farmacocinética , Cefalosporinas/administração & dosagem , Cefalosporinas/sangue , Cefalosporinas/farmacocinética , Feminino , Fluoroquinolonas , Humanos , Recém-Nascido , Injeções Intravenosas , Moxifloxacina , Ofloxacino/administração & dosagem , Ofloxacino/farmacocinética , Gravidez , Complicações Infecciosas na Gravidez/prevenção & controle , Quinolinas/administração & dosagem , Quinolinas/farmacocinéticaRESUMO
An HPLC method for the separation of seven cephalosporins [Cefepime (CEP), ceftazidime (CTA), ceftizaxime (CTI), ceftriaxone (CTR), cefotaxime (COT), cefixime (CIX) and cefoperazone (COP)] in human plasma and amniotic fluid has been developed. Optimization of the chromatographic method was performed in three steps: a series of initial experiments followed by two sets of experiments based on different experimental designs. The initial experiments were performed to decide the basic analytical requirements of the method. Then screening experiment fractional factorial design was used in order to decrease the number of parameters by eliminating parameters which having insignificant effect on responses. The parameters having significant effect were further optimized through a full factorial design. Having studied two responses (retention times and resolutions), a desirability function that assess the responses together, was used to find experimental conditions where the system generated desirable results. The desirable results were obtained with XTerra C18 (250 mm x 4.6mm, 5 microm i.d.) column, 40 mM phosphate buffer, pH 3.2, 18% MeOH, 0.85 mL min(-1) flow rate and 32 degrees C column temperature. Gradient elution with MeOH was applied. A simple and efficient solid-phase extraction was applied for the preparation of plasma and amniotic fluid samples. The validation parameters of the method were evaluated in accordance with ICH guideline. The method validated was applied to the analysis of CEP and COP in maternal venous, fetal venous and fetal arterial plasma, and to the analysis of CIX in maternal venous plasma and amniotic fluid.
Assuntos
Líquido Amniótico/química , Cefalosporinas/análise , Cefalosporinas/sangue , Cromatografia Líquida de Alta Pressão/métodos , Calibragem , Cefepima , Cefixima/análise , Cefixima/sangue , Cefixima/química , Cefoperazona/análise , Cefoperazona/sangue , Cefoperazona/química , Cefotaxima/análise , Cefotaxima/sangue , Cefotaxima/química , Ceftazidima/análise , Ceftazidima/sangue , Ceftazidima/química , Ceftizoxima/análise , Ceftizoxima/sangue , Ceftizoxima/química , Ceftriaxona/análise , Ceftriaxona/sangue , Ceftriaxona/química , Cefalosporinas/química , Estabilidade de Medicamentos , Feminino , Humanos , Estrutura Molecular , Gravidez , Reprodutibilidade dos Testes , TemperaturaRESUMO
OBJECTIVE: To establish a new high performance liquid chromatography (HPLC) method for determining the concentration of cefazolin, cefradine, cefoperazone and cefotaxime in blood and urine, as well as to investigate its applicability. METHODS: Protein in blood and urine was precipitated directly by acetonitrile with acetanilide was used as the internal standard using Agilent Zorbax SB-Aq column (250 mm x 4.6 mm, 5 microm). The mixed solvents of water (triethylamine 0.12%, acetic acid 0.12%) and acetonitrile were used as the mobile phase to separate cephalosporins using gradient elution method at 1 mL/min (flow rate) and 254 nm (detection wavelength). RESULTS: The working curve of four cephalosporins showed a good correlation (r = 0.9993), with the detection limit up to 0.01 microg/mL. The recovery rate was more than 81.2%. CONCLUSION: This method is fast, easy and accurate. It is suitable for biological analysis of the 4 cephalosporins of the blood and urine in practical cases.
Assuntos
Antibacterianos/sangue , Antibacterianos/urina , Cefalosporinas/sangue , Cefalosporinas/urina , Cromatografia Líquida de Alta Pressão/métodos , Adulto , Cefazolina/sangue , Cefazolina/urina , Cefoperazona/sangue , Cefoperazona/urina , Cefotaxima/sangue , Cefotaxima/urina , Cefradina/sangue , Cefradina/urina , Toxicologia Forense , Humanos , Masculino , Sensibilidade e Especificidade , Manejo de EspécimesRESUMO
Cefoperazone and sulbactam are prescribed in combination and used in the treatment of moderate to severe bacterial infections. Serious anaphylaxis is a rare side effect. This report describes a fatal case of suspected anaphylaxis after intravenous administration of a combination of the two drugs. Heart blood was analyzed for cefoperazone by protein precipitation with acetonitrile and by liquid-liquid precipitation for sulbactam after protein precipitation with aqueous acetonitrile, followed by tandem mass spectrometry in the product ion scan mode for identification and by liquid chromatography mass spectrometry in the selected ion monitoring mode for quantitation. Calibration curves for cefoperazone and sulbactam were linear over the range 0.07 to 1.93 and 0.046 to 0.914 microg/ml respectively. The decedent's blood concentrations of cefoperazone and sulbactam were 0.368 and 0.143 microg/ml respectively. As these concentrations were below concentrations reported after single dosing studies and below those considered to be minimally inhibitory, death was presumed to have been caused by hypersensitivity and not an overdose. In conclusion, this procedure is useful for detecting and quantitating cefoperazone and sulbactam in postmortem blood and may be useful in the evaluation of anaphylaxis.
Assuntos
Anafilaxia/induzido quimicamente , Antibacterianos/efeitos adversos , Cefoperazona/efeitos adversos , Cromatografia Líquida , Espectrometria de Massas , Sulbactam/efeitos adversos , Adulto , Antibacterianos/sangue , Cefoperazona/sangue , Toxicologia Forense , Humanos , Infusões Intravenosas , Masculino , Sulbactam/sangueRESUMO
We present a method for determining two cephalosporins (cefoperazone and ceftiofur) in plasma by on-line solid-phase extraction (SPE)-capillary zone electrophoresis (CZE) with a T-split interface. Using this interface, a part of the SPE elution plug containing the cephalosporins is injected while the rest of the sample is flushed to waste. SPE was carried out using a C(18) micro-precolumn and the cephalosporins presented good retention properties with breakthrough volumes above 1 ml. Using a desorption volume of 426 nl of acetonitrile, recoveries were 75 and 90%, for cefoperazone and ceftiofur, respectively. The resulting elution volume was about 1.8 microl. A deproteinization step was included prior to SPE for the analysis of plasma samples with recoveries of 90 and 57% for cefoperazone and ceftiofur, respectively. With UV detection at 254 nm, linear relationships between the injected concentration and peak area was measured between 10 and 500 ng ml(-1) for standards, and 200 and 1500 ng ml(-1) for plasma samples. Intra-day (n=5) and inter-day (n=5) peak area repeatability were lower than 12% RSD. The detection limits obtained for spiked plasma (100 ng ml(-1) cefoperazone and ceftiofur) are sufficient for applying the method to pharmacokinetic studies.
Assuntos
Antibacterianos/sangue , Cefoperazona/sangue , Cefalosporinas/sangue , Eletroforese Capilar/métodos , Animais , Bovinos , Humanos , Reprodutibilidade dos Testes , Espectrofotometria UltravioletaRESUMO
Acidity constants of six cephalosporin antibiotics, cefalexin, cefaclor, cefadroxil, cefotaxim, cefoperazon and cefoxitin are determined using capillary zone electrophoresis (CZE) and pH-potentiometric titrations. Since CZE is a separation method, it is not necessary for the samples to be of high purity and known concentration because only mobilities are measured. The effect on determination of dissociation constants of different matrices (serum, 0.9% NaCl, fermentation matrix) was examined. The advantages of CZE can be utilized in those fields where potentiometry has limitations (sample quantity, solubility, purity, simultaneous determinations), although pK(a) values that are close to each other can be determined by potentiometry with more accuracy.
Assuntos
Antibacterianos/química , Cefalosporinas/química , Eletroforese Capilar/métodos , Antibacterianos/análise , Cefaclor/análise , Cefaclor/sangue , Cefaclor/química , Cefadroxila/análise , Cefadroxila/sangue , Cefadroxila/química , Cefoperazona/análise , Cefoperazona/sangue , Cefoperazona/química , Cefotaxima/análise , Cefotaxima/sangue , Cefotaxima/química , Cefoxitina/análise , Cefoxitina/sangue , Cefoxitina/química , Cefalexina/análise , Cefalexina/sangue , Cefalexina/química , Cefalosporinas/análise , Cefalosporinas/sangue , Eletroforese Capilar/instrumentação , Concentração de Íons de Hidrogênio , Cinética , Estrutura Molecular , Potenciometria/instrumentação , Potenciometria/métodosRESUMO
A fully validated simple, sensitive and selective square-wave stripping voltammetry procedure was described for the trace quantification of cefoperazone in bulk form, formulations and human serum/plasma. The procedure was based on reduction of the adsorbed drug onto a hanging mercury drop electrode. The procedural conditions were optimized as: frequency=60Hz, scan increment=8mV, pulse amplitude=25mV, preconcentration potential=-0.3V (versus Ag/Ag/KCl(s)), preconcentration duration=60-150s and an acetate buffer of pH 4.2 as a supporting electrolyte. A limit of detection of 4.5x10(-10)M and a limit of quantitation of 1.5x10(-9)M bulk cefoperazone were achieved following preconcentration of the drug onto the hanging mercury drop electrode for 150s. The proposed square-wave adsorptive cathodic stripping voltammetric procedure was successfully applied for trace quantification of cefoperazone in human serum and plasma. The achieved limits of detection and quantitation of the drug in human serum were 6x10(-10)M (0.375ngml(-1)) and 2x10(-9)M (1.250ngml(-1)), respectively. The pharmacokinetic parameters of cefoperazone in plasma of hospitalized volunteers were successfully estimated.
Assuntos
Antibacterianos/farmacocinética , Cefoperazona/farmacocinética , Eletroquímica/métodos , Antibacterianos/análise , Antibacterianos/sangue , Antibacterianos/química , Cefoperazona/análise , Cefoperazona/sangue , Cefoperazona/química , Eletrodos , Humanos , Masculino , Mercúrio , Valores de Referência , Reprodutibilidade dos TestesRESUMO
A high performance frontal analysis(HPFA) method was developed to determine the unbound concentration of drugs in drug-human serum albumin (HSA) mixture under binding equilibrium. The sample was injected directly onto an internal-surface reversed-phase silica column (ISRP). The mobile phase was 67 mmol/L phosphate buffer (pH 7.4, I = 0.17 mol/L). When a large volume of sample solution under drug-HSA binding equilibrium was directly injected, the drug was eluted as a trapezoidal peak with a plateau, and the drug concentration in this region was the same as that of the unbound drug in the sample solution. The eluate of plateau region was collected and a small volume was injected onto a reversed-phase HPLC column. This HPFA-HPLC method was employed in the determination of unbound concentration in both ketoprofen (KP)-HSA and cefoperazone (CP)-HSA mixtures. The unbound concentrations of drugs obtained by using HPFA-HPLC were compared with those determined with ultrafiltration-HPLC. The effects of sample volume and flow rate of mobile phase on the plateau formation were investigated. It was found that the minimum injection volume to achieve a trapezoidal peak varied with drugs. The flow rate showed no effect on the trapezoidal peak formation. The unbound concentrations of KP and CP obtained were about the same by using HPFA-HPLC or ultrafiltration-HPLC and precisions were similar for both methods.
Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Albumina Sérica/metabolismo , Cefoperazona/sangue , Humanos , Cetoprofeno/sangue , Ligação Proteica , Proteínas/metabolismo , UltrafiltraçãoRESUMO
A sensitive microbore HPLC method was developed for the simultaneous determination of unbound cefoperazone in rat blood and brain using microdialysis. Two microdialysis probes were inserted into the jugular vein/right atrium and brain striatum of Sprague-Dawley rats. Cefoperazone (50 mgkg(-1), i.v.) was then administered via the femoral vein. Blood and brain dialysates were collected and eluted with a mobile phase containing methanol-100 mM monosodium phosphoric acid (30:70, v/v, pH 5.5). The wavelength of the UV detector was set at 254 nm. The detection limit of cefoperazone was 20 ng mL(-1). Isocratic separation of cefoperazone was achieved within 10 min. The intra- and inter-assay accuracy and precision of the analyses were < or =10% in the range of 0.05-10 microg mL(-1). The ratio of the area under the concentration curve of cefoperazone in rat brain and blood was estimated to be about 7-8%. It is concluded that cefoperazone is capable of penetrating the blood-brain barrier.
Assuntos
Encéfalo/metabolismo , Cefoperazona/farmacocinética , Cefalosporinas/farmacocinética , Animais , Área Sob a Curva , Barreira Hematoencefálica , Cefoperazona/sangue , Cefalosporinas/sangue , Cromatografia Líquida de Alta Pressão , Injeções Intravenosas , Masculino , Microdiálise , Ratos , Ratos Sprague-DawleyRESUMO
The pharmacokinetics and bioavailability of cefoperazone (CPZ) were studied following intravenous (IV) and intramuscular (IM) administration of single doses (30 mg/kg) to horses. Concentrations in serum, urine and synovial fluid samples were measured following IV administration. CPZ concentrations in serum, synovial fluid and spongy bone samples were measured following IM administration. After IV administration a rapid distribution phase (t1/2 (alpha): 4.22 +/- 2.73 min) was followed by a slower elimination phase (t1/2(beta) 0.77 +/- 0.19 h). The apparent volume of distribution was 0.68 +/- 0.10 L/kg. Mean synovial fluid peak concentration was 5.76 +/- 0.74 microgram/mliter. After IM administration a bioavailability of 42.00 +/- 5.33% was obtained. Half-life of absorption was 2.51 +/- 0.72 min and t1/2(beta) was 1.52 +/- 0.15 h. The mean synovial fluid and spongy bone peak concentrations at 2 h after IM administration were 2.91 +/- 0.85 microgram/mliter and 5.56 +/- 0.70 microgram/mliter, respectively.