title: Analysis of ochratoxin A in milk after direct immunoaffinity column clean-up by high-performance liquid chromatography with fluorescence detection.
abstract: The present work describes a new analytical method for direct immunoaffinity column clean-up of ochratoxin A (OTA) in milk samples followed by determination of the toxin using high-performance liquid chromatography with fluorescence detection (HPLC-FD). Two different immunoaffinity cartridges (IAC) were investigated, and Ochraprep columns were chosen because they showed the best results. An average recovery of 89.8% and a mean RSD of 5.8% for artificially contaminated cow's milk in the range of 5-100 ng/L were attained. The calculated limit of detection (LOD) and limit of quantitation (LOQ) were as low as 0.5 and 5 ng/L, respectively. This new easy and fast method avoids a previous liquid-liquid extraction step and therefore the use of toxic chlorinated solvents. Chromatograms of the final extracts were clean and OTA could be easily detected at a retention time of 8.4 min without interferences. To assess the presence of the toxin in cow's milk eight samples of skimmed and four samples of whole milk were analysed and OTA was not detected over the established detection limit.
authors.affiliation: Instituto de Productos Lácteos de Asturias (IPLA-CSIC), Carretera de Infiesto s/n, 33300 Villaviciosa, Asturias, Spain.
authors.forename: Victoria
authors.initials: V
authors.lastname: Bascarán
authors.forename: Alma Hernández
authors.initials: AH
authors.lastname: de Rojas
authors.forename: Paloma
authors.initials: P
authors.lastname: Chouciño
authors.forename: Teresa
authors.initials: T
authors.lastname: Delgado
mesh_terms: D053000:Analytic Sample Preparation Methods; D000818:Animals; D001230:Aspergillus / Q000737:chemistry; D002273:Carcinogens / Q000032:analysis*; D002417:Cattle; D005591:Chemical Fractionation / Q000379:methods; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D005504:Food Analysis / Q000379:methods; D005506:Food Contamination / Q000032:analysis*; D008892:Milk / Q000737:chemistry*; D009793:Ochratoxins / Q000032:analysis*; D010407:Penicillium / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence