Desenvolvimento e validação de metodologia analítica por clae em fase reversa para a determinação do perfil cromatográfico do extrato etanólico das cascas do caule de aspidosperma nitidum benth (apocynaceae)

Abstract

Aspidosperma nitidum Benth belonging to the Apocynaceae, popularly known as carapanaúba or peroba, widely used in the Amazon region, especially in the relief of symptoms of malaria, having as striking feature of the gener the great presence of indol alkaloids. Despite the widespread use of herbal folk medicine, problems as identification and standardization of the species are quite common. In this context, the present study aimed at the development and validation of an analytical method using high performance liquid chromatography, in order to determine the chromatographic profile of the ethanol extract of the stem bark of Aspidosperma nitidum. The ethanol extract was obtained by maceration and dried extract by freeze. A qualitative evaluation of chemically ethanol extract species showed the presence of alkaloids. The methodology was developed in a liquid chromatograph with high efficiency photodiode array detector, using reverse phase separation system. The validation procedure was carried out by the method of internal standardization, using the 1.8-dihidroxiantraquinona as standard. The calibration curves showed linearity (r ²> 0.99) in a wide range of concentration of the extract and standard, 62.5 to 2000 μg / mL and 15 to 500 μg / mL respectively. The test presented repeatability and reproducibility above 95%, since the coefficient of variation obtained from both assays are below 5%. The method showed good accuracy, which was shown by average recovery of 106% with a coefficient of variation of 2,2%. For all that, the chromatographic method developed and validated proved simple, linear, reproducible and accurate, It can be used therefore for determining the chromatographic profile of the ethanol extract of the stem bark Aspidosperma nitidum.