Desenvolvimento de mÃtodos para determinaÃÃo de microcistina-lr em Ãgua

AUTOR(ES)
DATA DE PUBLICAÇÃO

2003

RESUMO

In the search for methods for microcystins analysis that are easy to use, low cost and with sufficient high sensitivity to following the recommendations of the World Health Organization âWHO and the Brazilian legislation, was developed a specific dot blot assay. This dot blot imnunoassy is for visual determination of microcystin-LR in purified and sufarce water (lake). Concentration as 0.16 up to 10.0 μg/l of microcystin-LR in water were tested. Samples concentration or clean-up was not necessary for the water analysis. A specific software for membranes reading dot was developed, allowing a much better characterization of the dot with the attainment of similar analytical curves obtaining troughout the analyses by enzyme linked imunoassay - ELISA. With the computerized dot blot technique using a conventional scanner, coefficient of correlation of 0.9551 to purified water and 0.8402 to surface water were observed in the range of 0.16 at 1.6 μg/l. For ELISA, the value of ârâ were 0.9713 and 0.8316, respectively. Quantification in the range of 0.16 at 10.0 μg/l was carried through, however with lesser correlation between toxin concentration and intensity of pixels. The computerized analysis allowed to observe a good correlation between concentration and intensity of pixels using extract of toxic strain Microcystis aeruginosa - NPJB-1. This software was used for the determination of the bovine serum albumin - BSA in the range of 0.0 at 78.0 μg/mL (r = 0.9868) when stained with amidoblack solution. In order to get a new fluorescent labeled compound to the analysis of microcystin-LR in low concentrations, a complex of microcystin-LR-Terbium criptate was synthecized. The complex formation was followed by high performance liquid chromatography - HPLC and the ELISA, proteic reaction and luminescence spectrum utilized to confirme the conjugation. In order to lookinkg for new methods for microcystin analysis the formatoin of the transistion 5D0 Ã 7F2 of a IgG complex antimicrocystin-LR-KLH- Europium cryptate was monitored. Superabsorbent acrylic acid / polyacrilamyde gel was used as support. The emission spectrum of free IgG, europium cryptate and IgG-Europium cryptate complex had been monitored. The emission spectrum in the presence of IgG had presented a specific change in the transistion

ASSUNTO(S)

ciencias biologicas oms microcystinas cianobactÃrias

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