title: Validation of a confirmatory analytical method for the determination of aflatoxins B₁, B₂, G₁ and G₂ in foods and feed materials by HPLC with on-line photochemical derivatization and fluorescence detection.
abstract: A sensitive and selective analytical method for the simultaneous separation and quantitative determination of aflatoxins B1, B2, G1 and G2 in foodstuffs and materials for feed has been validated. The method is based on high performance liquid chromatography with on-line post-column photochemical derivatization and fluorescence detection. The chromatographic separation of aflatoxins was accomplished using a C18 column eluted with an isocratic mobile phase consisting of water, methanol and acetonitrile. The sample preparation required a simple extraction of aflatoxins with MeOH/H2O (80:20, v/v) and a purification step by immunoaffinity column cleanup. The total analysis time, including sample preparation and chromatographic separation, did not exceed 40 min with a run time of 10 min. The on-line photochemical derivatization ensures better results in terms of simplicity, sensitivity and reproducibility with respect to chemical derivatization techniques, and provides an increase of the peak resolution and an extent of automation in comparison with the electrochemical ones. The procedure for the determination of aflatoxins in food samples and cereals for animal consumption was extensively validated following Regulation (EC) No. 882/2004. Detection limits in wheat bran samples of 0.08 μg kg1 for AFB1, 0.02 μg kg1 for AFB2, 0.16 μg kg1 for AFG1 and 0.04 μg kg1 for AFG2 were attained. The method allows high recovery with mean values ranging from 72 to 94% and it satisfies the necessary requirements for sensitivity, linearity, selectivity, precision and ruggedness, demonstrating the conformity of the method with provisions of Regulation (EC) No. 401/2006.
authors.affiliation: Istituto Zooprofilattico Sperimentale della Puglia e della Basilicata, Via Manfredonia 20, 71100 Foggia, Italy.
authors.forename: Marilena
authors.initials: M
authors.lastname: Muscarella
authors.forename: Marco
authors.initials: M
authors.lastname: Iammarino
authors.forename: Donatella
authors.initials: D
authors.lastname: Nardiello
authors.forename: Sonia Lo
authors.initials: SL
authors.lastname: Magro
authors.forename: Carmen
authors.initials: C
authors.lastname: Palermo
authors.forename: Diego
authors.initials: D
authors.lastname: Centonze
authors.forename: Domenico
authors.initials: D
authors.lastname: Palermo
mesh_terms: D000348:Aflatoxins / Q000737:chemistry*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005453:Fluorescence; D005504:Food Analysis / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D055668:Photochemical Processes; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D000348:Aflatoxins