RESUMO
A quantitative analysis of polyamines in urine and serum by liquid chromatography-tandem mass spectrometry (LC-MS/MS) is described. The polyamines were carbamylated with isobutyl chloroformate, extracted with diethyl ether under pH 9.0, and analyzed by LC-MS/MS with single reaction monitoring mode. The limit of quantification was 1 ng/mL based on a signal-to-noise ratio>3, and the correlation coefficient (r2) for the calibration curves was >0.99 for both urine and serum samples. The present method was applied to urine and serum samples from 30 breast cancer patients and 30 normal female controls. There was no significant difference in the urinary polyamine levels between breast cancer patients and controls. However, 1,3-diaminopropane, putrescine, spermine and N-acetylspermidine levels in serum increased in breast cancer patients. These four serum polyamines may be a good index to study both production and metabolism of polyamines, and a useful tool in assessment of the polyamine status of breast cancer patients.
Assuntos
Biomarcadores Tumorais/sangue , Neoplasias da Mama/sangue , Cromatografia Líquida de Alta Pressão , Poliaminas/sangue , Poliaminas/urina , Espectrometria de Massas por Ionização por Electrospray , Espectrometria de Massas em Tandem , Biomarcadores Tumorais/urina , Neoplasias da Mama/química , Neoplasias da Mama/urina , Calibragem , Carbamatos/análise , Carbamatos/química , Fracionamento Químico , Técnicas de Laboratório Clínico , Diaminas/normas , Feminino , Formiatos/química , Humanos , Poliaminas/química , Padrões de Referência , Reprodutibilidade dos Testes , Sensibilidade e EspecificidadeRESUMO
The mycobactericidal and tuberculocidal activities of Korsolex AF against Mycobacterium tuberculosis, Mycobacterium avium-Mycobacterium intracellulare (MAI), Mycobacterium kansasii and Mycobacterium chelonae were determined using quantitative suspension and carrier tests. The effects of organic load and hard water were also considered. A clinical isolate of MAI was the most resistant of the four test organisms. A 2% solution had good mycobactericidal and tuberculocidal activities after 30 min of exposure. Although further evaluation using European standard tests is necessary, we conclude that Korsolex AF appears to be a promising product for the disinfection of hospital instruments contaminated with mycobacteria.
Assuntos
Aminas/normas , Detergentes/normas , Diaminas/normas , Desinfetantes/normas , Contaminação de Equipamentos/prevenção & controle , Mycobacterium/efeitos dos fármacos , Aminas/química , Cacau , Óleo de Coco , Contagem de Colônia Microbiana , Detergentes/química , Diaminas/química , Desinfetantes/química , Desinfecção/métodos , Desinfecção/normas , Avaliação Pré-Clínica de Medicamentos , Farmacorresistência Bacteriana , Europa (Continente) , Humanos , Testes de Sensibilidade Microbiana , Complexo Mycobacterium avium/efeitos dos fármacos , Mycobacterium chelonae/efeitos dos fármacos , Mycobacterium kansasii/efeitos dos fármacos , Mycobacterium tuberculosis/efeitos dos fármacos , Óleos de Plantas/química , Soluções , Suspensões , Fatores de TempoRESUMO
Glycosaminoglycans are identified in tissue sections by various histochemical techniques including staining with alcian blue and its analogues, such as cuprolinic blue and cupromeronic blue, or with high and low iron diamine methods. The variation in staining results in particularly confusing in the case of alcian blue, where not only are several different brands of alcian blue available but also several different staining protocols are used. If the results obtained by these techniques are compared, they often do not match. We have developed a dot blot technique for quality control of glycosaminoglycan histochemistry to standardize the staining protocols. This staining technique enables histochemists to test particular batches of alcian blue or its analogues for selective glycosaminoglycan staining, thus improving control of histochemical results. The results obtained using the dot blot assay indicate that it is necessary to test each batch of dye individually to obtain valid results in glycosaminoglycan histochemistry.
