RESUMO
A fucomannogalactan (FMG-Am) and a (1â3), (1â6)-linked ß-D-glucan (ßGLC-Am) were isolated from Amanita muscaria fruiting bodies. These compounds' structures were determined using mono- and bi-dimensional NMR spectroscopy, methylation analysis, and controlled Smith degradation. FMG-Am was shown to be a heterogalactan formed by a (1â6)-linked α-D-galactopyranosyl main chain partially substituted at O-2 mainly by α-L-fucopyranose and a minor proportion of ß-D-mannopyranose non-reducing end units. ßGLC-Am was identified as a (1â3)-linked ß-D-glucan partially substituted at O-6 by mono- and a few oligosaccharide side chains, which was confirmed after controlled Smith degradation. Both the homo- and heteropolysaccharide were evaluated for their anti-inflammatory and antinociceptive potential, and they produced potent inhibition of inflammatory pain, specifically, 91±8% (30 mg kg(-1)) and 88±7% (10 mg kg(-1)), respectively.
Assuntos
Amanita/química , Galactanos/química , Galactanos/farmacologia , Glucanos/química , Glucanos/farmacologia , Dor/complicações , Dor/tratamento farmacológico , Animais , Feminino , Formaldeído/efeitos adversos , Carpóforos/química , Galactanos/isolamento & purificação , Galactanos/uso terapêutico , Glucanos/isolamento & purificação , Glucanos/uso terapêutico , Inflamação/complicações , Masculino , Camundongos , Peso Molecular , Nociceptividade/efeitos dos fármacos , Dor/induzido quimicamente , Solubilidade , Relação Estrutura-AtividadeRESUMO
Amanitins are toxins found in species of the mushroom genera Amanita, Lepiota and Galerina. Intoxication after ingestion of these mushrooms can be fatal with an estimated 20% of mortality rate. An early diagnosis is necessary in order to avoid invasive and expensive therapy and to improve patient's prognosis. In this paper, a Capillary Zone Electrophoresis method was developed and validated to determine alpha- and beta-amanitin in urine in less than 7 min using 5 mM, pH 10 borate buffer as background electrolyte. The separation conditions were: capillary: 75 microm I.D., 41 cm effective length, 48 cm total length, 25 degrees C, 20 KV and PDA detection at 214 nm. Sample treatment for analysis only required urine dilution in background electrolyte. The method was validated following established criteria and was found to be selective, linear in the range 5-100 ng/ml. Intra- and inter-day precision and accuracy were within required limits. Limit of detection (LOD) and limit of quantification (LOQ) were 1.5 and 5 ng/ml, respectively. Eight urine samples from suspected cases of intoxication with amanitins were analyzed after 2 years of storage at -20 degrees C, and beta-amanitin was determined in two samples with concentrations of 53 and 65 ng/ml, respectively. The method here described includes the use of non-aggressive reagents to the capillary or the system and is the first Capillary Electrophoresis method used to determine amanitins in clinical samples.