Catecolaminas do aparelho reprodutor de ratos após simpatectomia química: metodologia analítica e comparação entre as variedades Wistar e Sprague-Dawley
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Data
2007-02-08
Autores
Silva, Daniela Alessandra Fossato da [UNESP]
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Universidade Estadual Paulista (Unesp)
Resumo
O tratamento com baixas doses de guanetidina provoca desnervação seletiva em órgãos sexuais acessórios de ratos Wistar e Sprague-Dawley, devido à depleção da noradrenalina nas terminações nervosas autonômicas simpáticas. O modelo experimental proposto possui semelhanças com casos de homens acometidos por lesão na medula espinhal. O objetivo deste trabalho foi investigar as concentrações de catecolaminas em órgãos reprodutores de ratos Wistar e Sprague-Dawley, após simpatectomia química por guanetidina e período de suspensão do tratamento, através do desenvolvimento e validação de um método analítico para a determinação simultânea de noradrenalina e adrenalina, por Cromatografia Líquida de Alta Eficiência com Detecção Eletroquímica. As amostras foram obtidas de ratos machos das variedades Wistar e Sprague- Dawley, controle e submetidos ao tratamento (via intraperitonial) com guanetidina, nas doses de 10mg/kg, durante 30 dias e 6,25mg/kg durante 21 dias, respectivamente. Foi utilizado ácido perclórico 0,4 mol L-1 no procedimento de extração das catecolaminas. Em seguida as seguintes etapas foram realizadas: avaliação do tempo de extração, ensaios de recuperação (empregando o método de calibração adição padrão) e determinação de noradrenalina e adrenalina nas amostras reais. O melhor rendimento de extração da noradrenalina foi em 90 min na cauda do epidídimo e 60 min no ducto deferente. A extração manteve-se constante a partir de 30 min para a adrenalina nas duas amostras de tecidos. O emprego do ácido perclórico 0,4 mol L-1 e o método cromatográfico com detecção eletroquímica permitiram uma boa extração e quantificação. A extração do analito resultou em valores de recuperação...
The treatment with low doses of guanethidine provokes selective denervation in accessory sexual organs of Wistar and Sprague-Dawley rats due to depletion of noradrenaline in the autonomic sympathetic nervous termination. The experimental model proposed has similarities with cases of men with spinal cord injury. The objective of this work was to investigate in the concentrations of catecholamines in reproductive organs of Wistar and Sprague-Dawley rats, after chemical sympathectomy with guanethidine and a period of suspension of the treatment, through the development and validation of an analytical method for the simultaneous determination of noradrenaline and adrenaline, by High Performance Liquid Chromatography with electrochemical detection. The samples were obtained from male rats of the varieties Wistar and Sprague-Dawley, control and those submitted to a treatment (intraperitonially) with guanethidine sulfate, at the doses of 10mg/kg, for 30 days and 6.25mg/kg for 21 days, respectively. Percloric acid 0.4 mol L-1 was used in the procedure of catecholamine extraction. Next, the following steps were performed: evaluation of the extraction time, assays of recovery (using the standard addition calibration method) and determination of noradrenaline and adrenaline in real samples. The best noradrenaline extraction yield was at 90 minutes for the cauda epididymis and 60 minutes for the vas deferens. The extraction was kept constant starting at 30 minutes for adrenaline in both tissue samples. The use of percloric acid 0.4 mol L-1 and the chromatographic method with eletrochemical detection allowed a good extraction and quantification. The extraction of the analyte resulted in recovery values between 56% and 114%, and coefficient of variation between 3.8% and 23%. The quantification limit of the method... (Complete abstract, click electronic access below)
The treatment with low doses of guanethidine provokes selective denervation in accessory sexual organs of Wistar and Sprague-Dawley rats due to depletion of noradrenaline in the autonomic sympathetic nervous termination. The experimental model proposed has similarities with cases of men with spinal cord injury. The objective of this work was to investigate in the concentrations of catecholamines in reproductive organs of Wistar and Sprague-Dawley rats, after chemical sympathectomy with guanethidine and a period of suspension of the treatment, through the development and validation of an analytical method for the simultaneous determination of noradrenaline and adrenaline, by High Performance Liquid Chromatography with electrochemical detection. The samples were obtained from male rats of the varieties Wistar and Sprague-Dawley, control and those submitted to a treatment (intraperitonially) with guanethidine sulfate, at the doses of 10mg/kg, for 30 days and 6.25mg/kg for 21 days, respectively. Percloric acid 0.4 mol L-1 was used in the procedure of catecholamine extraction. Next, the following steps were performed: evaluation of the extraction time, assays of recovery (using the standard addition calibration method) and determination of noradrenaline and adrenaline in real samples. The best noradrenaline extraction yield was at 90 minutes for the cauda epididymis and 60 minutes for the vas deferens. The extraction was kept constant starting at 30 minutes for adrenaline in both tissue samples. The use of percloric acid 0.4 mol L-1 and the chromatographic method with eletrochemical detection allowed a good extraction and quantification. The extraction of the analyte resulted in recovery values between 56% and 114%, and coefficient of variation between 3.8% and 23%. The quantification limit of the method... (Complete abstract, click electronic access below)
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Palavras-chave
Catecolaminas, Animais - Tecidos, Cromatografia líquida de alta eficiência
Como citar
SILVA, Daniela Alessandra Fossato da. Catecolaminas do aparelho reprodutor de ratos após simpatectomia química: metodologia analítica e comparação entre as variedades Wistar e Sprague-Dawley. 2007. 154 f. Tese (doutorado) - Universidade Estadual Paulista, Instituto de Química., 2007.