title: Development of a competitive ELISA for the evaluation of sunflower pollen in honey samples.
abstract: We report the development of a rapid, specific, and sensitive enzyme-linked immunoassay (ELISA) for the evaluation of sunflower pollen in honey as a method alternative to melissopalynology, which is considered the standard technique for the evaluation of floral origin of honey. Two 33-36 kDa proteins, identified as characteristic of sunflower pollen, were isolated and used as coating antigens in the competitive ELISA. We verified its analytical performance by evaluating reproducibility, specificity, and exactitude in relation to melissopalynology. The competitive ELISA developed during this work is able to quantify sunflower pollen in honey, with a detection limit of 10%, showing linear response between 10 and 90%. The method afforded low cross reactivity with honey from other floral origin, thus evidencing an adequate selectivity. We also observed a significant correlation (r = 0.975; p < 0.001) when the proposed ELISA was referenced to melissopalynology. Hence, we conclude that the competitive ELISA constitutes a valuable and feasible alternative for authentication of sunflower honey. This work opens the possibility to develop similar assays for other pollen types.
authors.affiliation: Universidad Nacional de Córdoba-CONICET, Facultad de Ciencias Químicas, Dto. Bioquímica Clínica-CIBICI, Medina Allende y Haya de la Torre, Ciudad Universitaria, 5000 Córdoba, Argentina.
authors.forename: María V
authors.initials: MV
authors.lastname: Baroni
authors.forename: Gustavo A
authors.initials: GA
authors.lastname: Chiabrando
authors.forename: Cristina
authors.initials: C
authors.lastname: Costa
authors.forename: Guillermina A
authors.initials: GA
authors.lastname: Fagúndez
authors.forename: Daniel A
authors.initials: DA
authors.lastname: Wunderlin
mesh_terms: D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006368:Helianthus / Q000737:chemistry*; D006722:Honey / Q000032:analysis* / Q000145:classification*; D011058:Pollen / Q000737:chemistry*; D012680:Sensitivity and Specificity