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Développement d'un support d'extraction sélective à base d'aptamères : synthèse, caractérisation et application à des matrices complexes

Abstract : The analysis of drugs in biological fluids (urine, plasma) at the trace level needs a sample clean-up before LC/MS analysis to limit the role of matrix effect. Despite their attractive features, classical sorbents used in Solid-Phase Extraction (SPE) often lead to the co-extraction of many interfering compounds. As an alternative to this lack of selectivity, selective materials based on a molecular recognition mechanism can be used. Immunosorbents (ISs) based on immobilized antibodies produced towards a target analyte, and molecularly imprinted polymers possessing specific cavities designed for a template molecule have already shown a high potential for the selective extraction of target analytes from complex matrices. Selective extraction based on a molecular recognition mechanism can also be obtained using aptamers immobilized onto a solid support. Aptamers are oligonucleotides able to bind a specific molecule with the same affinity as antibodies. Aptamers are less expensive to produce and present a higher stability than antibodies. An aptamer selected against cocaine was immobilized on various solid supports. These sorbents were characterized in terms of binding efficiency, specific and non specific retention as well as influence of the spacer length. To evaluate the ability of this oligosorbent to extract cocaine from complex samples, plasma and blood samples were treated by the support. The analysis of the samples treated by the oligosorbent led to a very clean chromatogram making the quantification of cocaine possible whereas peak co-elutions were observed when using conventional sample treatment techniques. This illustrated the improvement of the purification step by exploiting the high selectivity of aptamers. An oligosorbent was then synthesized following the same procedure with another aptamer, selected against ochratoxin A. A high retention was observed on this sorbent and allowed the selective extraction of ochratoxine A from wine. Moreover, the extraction procedures are more simple and easier to develop than for the molecularly imprinted polymers.
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Submitted on : Tuesday, November 23, 2010 - 2:07:32 PM
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  • HAL Id : pastel-00538840, version 1


Benjamin Madru. Développement d'un support d'extraction sélective à base d'aptamères : synthèse, caractérisation et application à des matrices complexes. Chimie-Physique [physics.chem-ph]. Université Pierre et Marie Curie - Paris VI, 2010. Français. ⟨pastel-00538840⟩



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