Immunoaffinity pre-concentration combined with on-column visual detection as a tool for rapid aflatoxin M1 screening in milk |
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| Authors: | Irina Yu. Goryacheva Makpal A. Karagusheva Carlos Van Peteghem Liberty Sibanda Sarah De Saeger |
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| Affiliation: | 1. Saratov State University, Chemistry Faculty, Department of Common and Inorganic Chemistry, Astrakhanskaya 83, 410012 Saratov, Russia;2. Ghent University, Faculty of Pharmaceutical Sciences, Laboratory of Food Analysis, Harelbekestraat 72, 9000 Ghent, Belgium;3. TOXI-TEST NV, Industrielaan 9a, 9990 Maldegem, Belgium;1. Aix Marseille Université, URMITE, UM63, CNRS 7278, IRD198, Inserm 1095, 13005 Marseille Cedex, France;2. UMR 912 SESSTIM, Inserm, IRD, Aix-Marseille Université, Faculté de Médecine, 27 Bd Jean Moulin, 13385 Marseille Cedex, France;3. Service de Chirurgie Cardiaque, Hôpital de la Timone, Marseille Cedex, France;1. Child & Family Research Institute, Vancouver, BC, Canada V5Z 4H4;2. University of British Columbia, Vancouver, BC, Canada V6T 1Z4;3. BC Children''s Hospital, Vancouver, BC, Canada V6H 3V4;4. MicroBiome Therapeutics, Broomfield, CO 80021, USA;1. Research Scholar, Department of ECE, Thiagarajar College of Engineering, Madurai, Tamil Nadu, India.;2. Department of ECE, Thiagarajar College of Engineering, Madurai, Tamil Nadu, India;3. Department of Oral & Maxillofacial Surgery, Best Dental Science College, Madurai, Tamil Nadu, India;1. Department of Natural Sciences, School of Arts and Sciences, Lebanese American University, Beirut, Lebanon;2. Department of Nutrition & Food Sciences, Faculty of Agricultural & Food Sciences, American University of Beirut, Beirut, Lebanon |
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| Abstract: | A non-instrumental rapid test was developed for the screening of aflatoxin M1 (AfM1) in milk at the 40 ng/L level. The method combines on one immunoaffinity gel layer, AfM1 pre-concentration and direct competitive immunoassay detection with visual evaluation. Aflatoxin B1-horse radish peroxidase (AfB1-HRP) and Sepharose 4B-immobilized anti-aflatoxin B1 monoclonal antibody with a 79% cross-reaction for AfM1 were used. The assay was performed in a standard column for solid phase extraction using a milk sample volume of 10 mL. Raw milk, pasteurized milk, milk powder, kefir and yogurt were analyzed with the developed test. It was shown that pasteurized milk could be analyzed without any sample preparation. The other types of milk were analyzed after centrifugation. For the assay procedure an aliquot of milk was flowed through the immunoaffinity gel layer in the tube by pushing the plunger of a syringe. Further, AfB1-HRP was applied and bound to free antibody binding sites. After addition of the chromogenic substrate the results were visually evaluated. Blue colour developed in case of negative milk samples while no colour appeared for positive samples. This method showed a false negative rate of only 2% and a high throughput (20–35 min for six samples). |
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