Characterization of oligosorbents and application to the purification of ochratoxin A from wheat extracts |
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Authors: | Wassim Hadj Ali Valérie Pichon |
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Institution: | 1. Department of Analytical and Bioanalytical Sciences and Miniaturization (LSABM), ESPCI ParisTech, UMR PECSA 7195 (CNRS-UPMC-ESPCI ParisTech), 10 rue Vauquelin, 75231, Paris Cedex 05, France
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Abstract: | The aim of this work was to optimize the preparation of an anti-ochratoxin A (OTA) oligosorbent (OS), a solid-phase extraction sorbent based on OTA aptamers covalently immobilized on sepharose. Different syntheses were carried out by modifying the side of the oligonucleotide chain bound to the sepharose, the length of the spacer arm between the aptamer and the sepharose and the amount of the aptamers introduced during the covalent grafting. Indeed, the capacity of OSs prepared using 3′- or 5′-amino-modified sequences with a C6 or a C12 was studied. In the best conditions, the concentration of aptamers sequence used during their grafting was increased and a capacity close to 40 nmol g?1 of OS was reached. The potential of the resulting OSs was also studied in pure media. For this, their selectivity was checked by comparing them to a control sorbent prepared without immobilizing aptamers. Extraction recoveries close to 100 % were obtained on all OSs, while no retention was observed on the control sorbent. OS does not demonstrate any cross-reactivity towards OTA metabolites, i.e., ochratoxin B and ochratoxin hydroquinone. The oligosorbent was finally applied to the clean-up of OTA from wheat sample extracts. Extraction recoveries were not affected by matrix interferences and the resulting chromatogram clearly highlights the selectivity of the sorbent that allows the removal of matrix components thus improving the reliability of the quantitation of OTA in real samples. |
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