The development and characterization of an anti-aflatoxin B1 (anti-AFB1) immunoaffinity monolithic disk is reported. Polyclonal anti-AFB1 was covalently immobilized in batch on an epoxy-activated monolithic Convective Interaction Media (CIM) disk (12mm×3mmi.d.) by a one-step reaction via epoxy groups of the polymer surface. 0.96 mgof antibody were immobilized and the binding capacity of the CIM diskwas determined by frontal analysis. The CIM diskwas coupled through a switching valve to a reversed-phase column, namely Chromolith Performance RP-18e.Afully automated HPLC method with fluorescence detection for the determination of aflatoxin B1 in aqueous solution was developed. The total analysis time with the integrated system is 46 min and the retention time of AFB1 is approximately 29 min. The binding capacity of the immunoaffinity disk was evaluated in terms of linearity, precision and accuracy of the extraction procedure. The immunoaffinity support was stable after repeated runs.
Development and integration of an immunoaffinity monolithic disk for the on-line solid-phase extraction and HPLC determination with fluorescence detection of aflatoxin B1 in aqueous solutions
CALLERI, ENRICA;MARRUBINI BOULAND, GIORGIO CARLO;BRUSOTTI, GLORIA;MASSOLINI, GABRIELLA;CACCIALANZA, GABRIELE
2007-01-01
Abstract
The development and characterization of an anti-aflatoxin B1 (anti-AFB1) immunoaffinity monolithic disk is reported. Polyclonal anti-AFB1 was covalently immobilized in batch on an epoxy-activated monolithic Convective Interaction Media (CIM) disk (12mm×3mmi.d.) by a one-step reaction via epoxy groups of the polymer surface. 0.96 mgof antibody were immobilized and the binding capacity of the CIM diskwas determined by frontal analysis. The CIM diskwas coupled through a switching valve to a reversed-phase column, namely Chromolith Performance RP-18e.Afully automated HPLC method with fluorescence detection for the determination of aflatoxin B1 in aqueous solution was developed. The total analysis time with the integrated system is 46 min and the retention time of AFB1 is approximately 29 min. The binding capacity of the immunoaffinity disk was evaluated in terms of linearity, precision and accuracy of the extraction procedure. The immunoaffinity support was stable after repeated runs.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
