Spectrophotometric determination of cefaclor in pharmaceutical preparations

A simple spectrophotometric method is proposed for the determination of cefaclor. The method involves alkaline hydrolysis of the drug in ammonia buffer solution at pH 10 to yield diketopiperazine-2,5-dione derivative and subsequent measurement at 340 nm. Beer's law is obeyed in the concentration range 1.8 - 55 mg/mL. The proposed method was successfully applied to the determination of cefaclor in pharmaceutical formulations.

Development and application of spectrophotometric method for the determination of cefaclor in pharmaceutical formulations

A simple, fast and sensitive spectrophotometric method for the determination of cefaclor in pharmaceutical raw and dosage forms based on reaction with ninhydrin is developed, optimized and validated. The purple color (Ruhemenn's purple) that resulted from the reaction was stabilized and measured at 560 nm. Beer's law is obeyed in the concentration range of 4-80 µg mL-1 with molar absorptivity of 1.42 × 10(5) L mole-1 cm-1. All variables including the reagent concentration, heating time, reaction temperature, color stability period, and cefaclor/ninhydrin ratio were studied in order to optimize the reaction conditions. No interference was observed from common pharmaceutical adjuvant. The developed method is easy to use, accurate and highly cost-effective for routine studies relative to HPLC and other techniques.

Spectrophotometric determination of cefaclor in pharmaceutical preparations

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Cathodic stripping voltammetric determination of cefaclor in pharmaceutical formulations

A sensitive method is described for the determination of cefaclor by cathodic stripping voltammetry at the hanging mercury drop electrode. cefaclor is accumulated at the electrode surface as a mercury salt, which is reduced at -0.67 V. The optimum accumulation potential and accumulation time were +0.15 V and up to 180 s, respectively. Linear calibration graphs were obtained between 3.9 mu g.L-1 to 39 mu g.L-1 and the limit of determination was evaluated to be 1.9 mu g.L-1. The method was applied successfully to the determination of cefaclor in pharmaceutical formulations.

Indirect polarographic and cathodic stripping voltammetric determination of cefaclor as an alkaline degradation product

Cefaclor is not reducible at a mercury electrode, but it can be determined polarographically and by cathodic stripping voltammetry as its initial alkaline degradation product which is obtained in high yield by hydrolysis of cefaclor in Britton-Robinson (B-R) buffer pH 10 at 50 degrees C for 30 min (reduction peak at pH 10, -0.70 V). Differential pulse polarographic calibration graphs are linear up to at least 1 x 10(-4) mol l(-1). Recoveries of 93% of the cefaclor (n = 3) were obtained from urine spiked with 38.6 mu g ml(-1) using this polarographic method with 1 ml urine made up to 10 ml with pH 10 buffer. Using cathodic stripping voltammetry and accumulating at a hanging mercury drop electrode at -0.2 V for 30 s, linear calibration graphs were obtained from 0.35 to 40 mu g ml(-1) cefaclor in B-R buffer pH 10. A relative standard deviation of 4.2% (eta = 5) was obtained, and the limit of detection was calculated to be 2.9 ng ml(-1). Direct determination of cefaclor in human urine (1 ml of urine was made up to 10 ml with pH 10 buffer) spiked to 0.39 mu g ml(-1) was made (recovery 98.6%). (C) 1999 Elsevier B.V. B.V. All rights reserved.

Allergy to betalactam antibiotics in children: a prospective follow-up study in retreated children after negative responses in skin and challenge tests.

BACKGROUND: Up to 10% of the patients in whom suspected betalactam hypersensitivity (HS) has been excluded by skin and challenge tests report suspected allergic reactions during subsequent treatments with the same or very similar betalactams. It has been suggested that the reactions may result from a resensitization induced by the challenge performed at the time of the allergological work-up. However, most patients did not undergo a second allergological work-up, to determine if the reactions resulted from betalactam HS or not. OBJECTIVES: We aimed to determine if children diagnosed nonallergic to betalactams have tolerated subsequent treatments with the initially suspected and/or other betalactams, and, in case of a reaction, if the reaction resulted from betalactam HS. Methods: We sent a questionnaire concerning the clinical history of their children to the parents of 256 children previously diagnosed nonallergic to betalactams. A second allergological work-up was performed in the children reporting suspected allergic reactions during subsequent treatments with the same and/or other betalactams. Skin tests were performed with the soluble form of the suspected (or very similar) betalactams and other betalactams from the same and other classes. Skin test responses were assessed at 15-20 min (immediate), 6-8 h (semi-late) and 48-72 h (late). Oral challenge (OC) was performed in children with negative skin tests, either at the hospital (immediate and accelerated reactions), or at home (delayed reactions). RESULTS: A response was obtained from 141 children (55.3%). Forty-eight (34%) of those children had not been treated with the betalactams for whom a diagnosis of allergy had been ruled out previously. Seven (7.5%) of the 93 children who had been treated again reported suspected allergic reactions. Skin tests and OC were performed in six of those children, and gave negative results in five children. In one child previously diagnosed nonallergic to amoxicillin associated with clavulanic acid, we diagnosed a delayed HS to clavulanic acid and a serum sickness-like disease to cefaclor. Thus, the frequency of reactions resulting from betalactam HS in children with negative skin and challenge tests is very low, and does not exceed 2.1% (2/93) if we consider that the child which refused a second allergological work-up is really allergic to betalactams. CONCLUSION: Our results in a very large number of children show that reactions presumed to result from betalactam HS are rare in children in whom the diagnosis of betalactam allergy has been ruled out previously. Moreover, they suggest that, as shown for the initial reactions, most of the reactions during subsequent treatments are rather a consequence of the infectious diseases for whom betalactams have been prescribed than a result of betalactam HS. Finally, they suggest that the risk of resensitization by OC is very low, and do not support the notion that skin testing should be repeated in children diagnosed nonallergic to betalactams.

Estudo voltamétrico de redissolução catódica de cefaloglicina em meio aquoso sobre eletrodo de mercúrio

Voltamogramas de redissolução catódica de cefaloglicina em meio ácido exibem uma única onda de redução em potenciais próximos da descarga do eletrólito (-1.0 V). Entretanto, o fármaco pode ser determinado indiretamente em meio alcalino (pH 10) através do complexo formado entre cefaloglicina e Hg(I) gerado na superfície do eletrodo sob potencial de acúmulo positivo (Eac= +0.20 V). Uma curva de calibração linear no intervalo de 0.122 a 1.22 mg/mL foi obtida para cefaloglicina em tampão B-R pH 10 utilizando-se tempo de acúmulo de 30 seg e potencial de acúmulo de +0.20 V. O método mostra alta reprodutibilidade e sensibilidade com desvio padrão relativo de 1,4 % para 5 determinações de 5x10-7 mol/L de cefaclor e limite de detecção de 5,17 ng/mL.

Caracterização da mastite subclínica em caprinos produzidos em sistema orgânico no Rio Grande do Sul

O leite e derivados são reconhecidos como veículos de patógenos para humanos, secundário a contaminação pós-ordenha ou de infecções do próprio animal, particularmente na mastite. Foi estudada a ocorrência de mastite e aspectos do manejo em cabras de três propriedades criadas em sistema orgânico. O exame clínico da glândula mamária de 64 cabras em diferentes períodos de lactação, não acusou a presença de mastite clínica. Entretanto, o Califórnia Mastitis Test (CMT) identificou 54 (22,7%) metades mamárias reagentes (+ ou ++). Foram colhidas 238 amostras de leite, das quais houve isolamento bacteriano em 37 (15,6%). Em apenas oito amostras houve coincidência entre o isolamento bacteriano e o resultado do CMT, indicando sensibilidade de 21,6% para este teste no diagnóstico de mastite subclínica em caprinos. Staphylococcus coagulase negativa (SCN) foi o microrganismo mais freqüente (83,8%). O teste de sensibilidade microbiana in vitro revelou resistência das linhagens de SCN ao cotrimoxazol (50%), ampicilina (48,1%), nitrofurantoína (7,7%), cefaclor (7,14%) e oxacilina (3,85%). Cefalotina, gentamicina, neomicina, estreptomicina e tetraciclina foram os antimicrobianos mais efetivos frente aos isolados. Não se evidenciou relação entre a ocorrência de mastite subclínica com a raça, a fase de lactação, sistema de ordenha ou qualidade da água utilizada nas propriedades.

Sobrevivência de microorganismos mesófilos e perfil físico-químico em estação de tratamento de dejetos suínos.

As variáveis físicas, químicas e microbiológicas de um sistema de lagoas interligadas para o tratamento de dejetos suínos foram avaliadas. O sistema, composto por sete lagoas em séries (duas anaeróbias, uma facultativa, uma com aeração mecânica e três aeróbias), está localizado no sul do Brasil e recebe dejetos de cerca de 4.000 matrizes e 30.000 suínos em crescimento e terminação. Foram realizadas 20 coletas quinzenais, em sete pontos ao longo do sistema de tratamento. Verificou-se a existência de diferença significativa (p<0,05) na quantidade de fosfato (PO4), nitrato (NO3), fósforo total (PT), sólidos totais (ST) e sólidos voláteis (SV) entre os pontos iniciais do sistema, os quais são anteriores ao tratamento propriamente dito, e as demais fases do processo. A maior redução dos parâmetros analisados ocorreu após as lagoas anaeróbias, havendo uma contínua diminuição dos mesmos no decorrer do processo (p<0,05). As reduções observadas foram de 97,5% para Demanda Bioquímica de Oxigênio (DBO), 97,2% para Demanda Bioquímica de Oxigênio (DQO), 74,8% PO4, 91,2% NO3, 70% PT, 77,4% ST, 86,7% SV, 99% de Coliformes Totais (CT) e 99% de Coliformes Fecais (CF) comparando-se os valores médios no início e no final do sistema. O sistema demostrou, ainda, ser eficaz no controle de Salmonella sp. Das 20 coletas realizadas, foi possível isolar Salmonella sp. em 13 coletas no ponto correspondente ao início do sistema de tratamento e em apenas uma no ponto final do mesmo. Ao lado disto, verificou-se modificação da microbiota mesófila aeróbia ao longo do sistema onde, no afluente predominaram microorganismos Gram negativos com características de enterobactérias e no efluente, cocos Gram positivos catalase negativos. Entretanto, não houve redução significativa no número de unidades formadoras de colônias de mesófilos aeróbios ao longo do sistema. Das 161 amostras de Salmonella Typhimurium e 186 amostras de Escherichia coli isoladas, determinou-se o perfil de resistência pelo método de difusão em ágar, usando 14 antimicrobianos. Observou-se resistência contra sulfonamida (99,5% e 100%), tetraciclina (97,3% e 99,4%), ampicilina (96,8% e 76,4%), estreptomicina (96,2% e 90,1%), sulfa/trimetoprima (95,2% e 84,5%), ácido nalidíxico (82,8% e 77,6%), cloranfenicol (70,4% e 29,2%), cefaclor (71,5% e 25,5%), neomicina (38,2% e 5%), gentamicina (37,1% e 6,2%), tobramicina (35,5% e 13,7%), ciprofloxacina (30,1% e 0%), amoxacilina/ácido clavulânico (11,8% e 5%) e amicacina (9,1% e 3,7%) para E. coli e Salmonella respectivamente, sendo que todas as amostras de Salmonella foram sensíveis à ciprofloxacina. A resistência a quatro ou mais antimicrobianos foi observada em 99,5% das amostras de E. coli e 94,5% das amostras de Salmonella. O padrão de multiresistência foi mantido ao longo do sistema, apesar de verificar-se uma tendência à menor resistência nas amostras de E. coli isoladas após a passagem pelas lagoas aeróbias. As amostras, tanto da afluente como do efluente do sistema, apresentaram grande variabilidade nos perfis de resistência.

Isolation of Salmonella enterica Serovar Enteritidis in Blue-Fronted Amazon Parrot (Amazona aestiva)

Avian salmonellosis is a disease caused by bacteria of the genus Salmonella that can cause three distinct diseases in birds: pullorum diseases, fowl typhoid, and paratyphoid infection. Various wildlife species are susceptible to infections by Salmonella, regardless of whether they live in captivity or freely in the wild. The present study verified the presence of Salmonella enterica serovar Enteritidis in three captive specimens of Amazona aestiva. The study involved a total of 103 birds undergoing rehabilitation to prepare for living in the wild, after having been captured from animal traffickers and delivered to the Centrofauna Project of the Floravida Institute in São Paulo, Brazil. This is the first report of Salmonella Enteritidis isolation in A. aestiva that originated from capture associated with animal trafficking; Salmonella was detected during the study by the serologic method of rapid serum agglutination on a plate with bacterial isolate. The antimicrobial profile exam of the isolated samples demonstrated sensitivity to ampicillin, cefaclor, ciprofloxacin, and cloranfenicol. The three samples also presented resistance to more than four antibiotics. The presence of the genes invA and spvC was verified by PCR technique and was associated with virulence and absence of class 1 integron, a gene related to antimicrobial resistance. The commercial antigen for pullorum disease was shown to be a useful tool for rapid detection in the screening of Salmonella of serogroup D(1) in Psittaciformes. New studies on Salmonella carriage in birds involved in trafficking must be performed to better understand their participation in the epidemiologic cycle of salmonellosis in humans and other animals.

Estudo voltamétrico de redissolução catódica de cefaloglicina em meio aquoso sobre eletrodo de mercúrio

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Micellar modification of drug stability: analysis of the effect of hexadecyltrimethylammonium halides on the rate of degradation of cephaclor

The intra- and intermolecular rates of degradation of cephaclor were determined with and without hexadecyltrimethylammonium bromide (CTABr). Micellar-derived spectral shifts were used to measure the association of the ionic forms as well as to determine the effect of CTABr on the apparent acid dissociation constant of the antibiotic. The rate of degradation of cephaclor increased with detergent and was salt sensitive. Micellar effects were analyzed quantitatively within the frame-work of the speudophase ion exchange model. All experimental data were fitted to this model which was used to predict the combined effects of pH and detergent concentration. Micelles increased the rate of OH- attack on cephaclor; most of the effect was due to the concentration of reagents in the micellar pseudophase. The intramolecular degradation was catalyzed 25-fold by micelles, and a working hypothesis to rationalize this effect is proposed. The results demonstrate that quantitative analysis can be utilized to assess and predict effects of detergents on drug stability.

Effect of micelles, microemulsions and other supramolecular aggregates on the hydrolysis of β-lactam antibiotics

A critical revision of the literature was made regarding the stability of β-lactam antibiotics in the presence of surfactants. The factors involved in the drug decomposition were analyzed in the development of the discussion. The analysis has indicated that some organized systems obtained from surfactants can be used to control rates and mechanisms of antibiotic decomposition. These organized systems can also be used to obtain specific information about the drug reactivity in a microenvironment similar to the site of pharmacological effect.

Padrão de sensibilidade de cepas de Enterobacter spp isoladas na região de Presidente Prudente - SP

Enterobacter spp. are considered important causing agents of infection, specially in hospitalized individuals. The natural resistance of these microorganisms and the great facility to develop resistance to new antimicrobial agents make this genus an important object of study. In this work, 176 strains isolated from various clinical samples were used from hospitalized patients (University Hospital Domingos Leonardo Cerávolo) and from clinic patients (Clinical Laboratory from Unoeste), both situated in Presidente Prudente - SP. E. cloacae (78.9%), E. aerogenes (7.9%) and E. (pantoea) agglomerans (3.9%) were the ones more frequently isolated. Eleven antimicrobial agents were tested by the disk diffusion method and around 90% of the strains presented resistance to the cephalotin, ampicillin and cefaclor. Strains of E. (pantoea) agglomerans presented wide profile of sensibility However one strain of E. cloacae presented resistance to all the antimicrobial agents. The antimicrobial agents with greater inhibitory activity were imipenem and cefepime, for this reason these antimicrobial agents could be the treatment of choice in emergencial therapeutic. This emergencial therapy can be applied with relative security, whereas the data obtained in this study show homogeneity in the profile of sensibility to these antimicrobial agents, independent of the infection site and from the isolated species. The ESBL enzyme could not be detected in no one of the strains by the double diffusion test.

Effect of hexadecyltrimethylammonium bromide-based microemulsions on the rate of decomposition of the beta-lactam antibiotic cephaclor

Microemulsions of hexadecyltrimethylammonium bromide (HTAB)/n-butanol/hexadecane/water catalyze the intramolecular degradation of cephaclor. The rate increase is a sensitive function of the microemulsion volume fraction and salt concentration. The effects of microemulsions, analyzed quantitatively using a pseudophase ion-exchange model, assumed that the extent of ion dissociation from the microemulsions varies with volume fraction. Comparison of micellar and microemulsion effects on the same reaction shows that microemulsions are less effective catalysts. Acceleration decreased significantly by increasing the relative proportion of n-butanol ratio in microemulsions and by addition of n-butanol in HTAB micelles. Comparison of the activation parameters of the reaction in aqueous solution, microemulsions, and micelles suggests that catalysis by both aggregates is driven mainly by entropic contributions.