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A vinyl sulfone clicked carbon dot-engineered microfluidic paper-based analytical device for fluorometric determination of biothiols

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Abstract

A microfluidic paper-based analytical device integrating carbon dot (CDs) is fabricated and used for a fluorometric off-on assay of biothiols. Vinyl sulfone (VS) click immobilization of carbon dots (CDs) on paper was accomplished by a one-pot simplified protocol that uses divinyl sulfone (DVS) as a homobifunctional reagent. This reagent mediated both the click oxa-Michael addition to the hydroxyl groups of cellulose and ulterior covalent grafting of the resulting VS paper to NH2-functionalized CDs by means of click aza-Michael addition. The resulting cellulose nanocomposite was used to engineer an inexpensive and robust microfluidic paper-based analytical device (μPAD) that is used for a reaction-based off-on fluorometric assay of biothiols (GSH, Cys, and Hcy). The intrinsic blue fluorescence of CDs (with excitation/emission maxima at 365/450 nm) is turned off via the heavy atom effect of an introduced iodo group. Fluorescence is turned on again due to the displacement of iodine by reaction with a biothiol. The increase in fluorescence is related to the concentration over a wide range (1 to 200 μM for GSH and 5–200 μM for Cys and Hcy, respectively), and the assay exhibits a low detection limit (0.3 μM for GSH and Cys and 0.4 μM for Hcy). The method allows for rapid screening and can also be used in combination with a digital camera readout.

Schematic representation of a μPAD based on click immobilized carbon dots and used for a reaction-based fluorometric off-on assay of biothiols. The intrinsic blue fluorescence of carbon dots is turned off via the heavy atom effect of an introduced iodo group and turned on by the displacement of this atom by reaction with a biothiol.

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Acknowledgments

This study was supported by projects from the Spanish MINECO (CTQ2017-86125 and CTQ2016-78754-C2-1-R). The project was also partially supported by the European Regional Development Funds (ERDF).

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Authors and Affiliations

  1. Department of Analytical Chemistry, Faculty of Sciences, University of Granada, Campus Fuentenueva s/n, 18071, Granada, Spain

    Inmaculada Ortiz-Gomez, Antonio Marín-Sánchez, Ignacio de Orbe-Payá, Luis Fermin Capitan-Vallvey & Alfonso Salinas-Castillo

  2. Unit of Excellence in Chemistry Applied to Biomedicine and the Environment, University of Granada, 18071, Granada, Spain

    Inmaculada Ortiz-Gomez, Mariano Ortega-Muñoz, Antonio Marín-Sánchez, Ignacio de Orbe-Payá, Fernando Hernandez-Mateo, Luis Fermin Capitan-Vallvey, Francisco Santoyo-Gonzalez & Alfonso Salinas-Castillo

  3. Department of Organic Chemistry, Biotechnology Institute, Faculty of Sciences, University of Granada, Campus Fuentenueva s/n, 18071, Granada, Spain

    Mariano Ortega-Muñoz, Fernando Hernandez-Mateo & Francisco Santoyo-Gonzalez

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  1. Inmaculada Ortiz-Gomez
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  2. Mariano Ortega-Muñoz
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  8. Alfonso Salinas-Castillo
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Correspondence to Alfonso Salinas-Castillo.

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Ortiz-Gomez, I., Ortega-Muñoz, M., Marín-Sánchez, A. et al. A vinyl sulfone clicked carbon dot-engineered microfluidic paper-based analytical device for fluorometric determination of biothiols. Microchim Acta 187, 421 (2020). https://doi.org/10.1007/s00604-020-04382-9

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