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Miniaturized DNA aptamer-based monolithic sorbent for selective extraction of a target analyte coupled on-line to nanoLC

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Abstract

A complete characterization of a novel target-specific DNA aptamer-based miniaturized solid phase extraction (SPE)-sorbent coupled on-line to nanoLC is presented. A miniaturized oligosorbent (mOS) was prepared via the in situ sol-gel synthesis of a hybrid organic-inorganic monolith in 100 μm i.d. capillary columns using tetraethoxysilane and 3-aminopropyltriethoxysilane as precursors, followed by covalent binding of a 5′-amino-modified DNA aptamer with a C12 spacer arm specific for a molecule of small molecular weight. Ochratoxin A (OTA), one of the most abundant naturally occurring mycotoxins, was chosen as model analyte to demonstrate the principle of such an approach. The mOS was coupled on-line to RP-nanoLC-LIF. Selective extraction of OTA on several mOSs was demonstrated with an average extraction recovery above 80 % when percolating spiked binding buffer and a low recovery on control monoliths grafted with a non-specific aptamer. Reproducibility of mOSs preparation was highlighted by comparing extraction yields. Otherwise, the mOSs demonstrated no cross-reactivity towards an OTA structural analogue, i.e., ochratoxin B. Due to the high specific surface area of the hybrid silica-based monolith, the coverage density of DNA aptamers covalently immobilized in the capillaries was very high and reached 6.27 nmol μL−1, thus leading to a capacity above 5 ng of OTA. This miniaturized device was then applied to the selective extraction of OTA from beer samples. It revealed to be effective in isolating OTA from this complex matrix, thus improving the reliability of its analysis at the trace level.

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Acknowledgments

This work was supported by the French National Research Agency (ANR Program CESA 2010, Mycodiag Project).

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Authors and Affiliations

  1. Department of Analytical, Bioanalytical Sciences and Miniaturization (LSABM)—UMR CBI 8231 (CNRS-ESPCI), ESPCI ParisTech, PSL Research University, 10 rue Vauquelin, 75231, Paris Cedex 05, France

    Fabien Brothier & Valérie Pichon

  2. Sorbonne Universités, UPMC, 4 Place Jussieu, 75005, Paris, France

    Valérie Pichon

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  1. Fabien Brothier
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  2. Valérie Pichon
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Correspondence to Valérie Pichon.

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Brothier, F., Pichon, V. Miniaturized DNA aptamer-based monolithic sorbent for selective extraction of a target analyte coupled on-line to nanoLC. Anal Bioanal Chem 406, 7875–7886 (2014). https://doi.org/10.1007/s00216-014-8256-z

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  • DOI: https://doi.org/10.1007/s00216-014-8256-z

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