Navegando por Autor "Ribeiro, Ricardo Marcelo Reche"

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    A comparison between enzyme immunoassay and HPLC for ochratoxin A detection in green, roasted and instant coffee
    (Instituto de Tecnologia do Paraná - Tecpar, 2007-03) Fujii, Simone; Ono, Elisabete Yurie Sataque; Ribeiro, Ricardo Marcelo Reche; Assunção, Fernanda Garcia Algarte; Takabayashi, Cássia Reika; Oliveira, Tereza Cristina Rocha Moreira de; Itano, Eiko Nakagawa; Ueno, Yoshio; Kawamura, Osamu; Hirooka, Elisa Yoko
    An indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) for ochratoxin A (OTA) detection in green, roasted and instant coffees was developed using anti-OTA monoclonal antibody. Immunological reagents prepared were OTA-BSA (4.76 μ g/mL), anti-OTA.7 MAb (2x10 3 -fold dilution) and HRP-anti IgG (10 3 -fold dilution). The detection limit was 3.73 ng OTA/g and correlation coefficients (r) between this immunoassay and high performance liquid chromatography were 0.98 for green coffee, 0.98 for roasted and 0.86 for instant. OTA levels detected by ic- ELISA were higher than by HPLC, with ELISA/HPLC ratio of 0.66 - 1.46 (green coffee), 0.96 - 1.11 (roasted) and 0.93 - 1.82 (instant). ELISA recoveries for OTA added to coffee (5 - 70 ng/g) were 81.53 % for green coffee, 46.73 % for roasted and 64.35 % for instant, while recoveries by HPLC were 80.54 %, 45.91 % and 55.15 %, respectively. Matrices interferences were minimized by samples dilution before carrying out the ELISA assay. The results indicate that MAb-based ic-ELISA could be a simple, sensitive and specific screening tool for OTA detection, contributing to quality and safety of coffee products.