Abstract
The oxidation of paper by periodate was investigated and systematically characterized by Fourier-transform infrared (FTIR) spectroscopy, scanning electron microscopy, X-ray diffraction, goniometry, and dynamic mechanical analysis. For the first time, in situ FTIR microscopy analysis was performed, yielding chemical images of carbonyl groups on the cellulose fibers. The enhancement of protein immobilization on oxidized paper was quantified by a colorimetric assay with Ponceau dye, demonstrating that 0.5-h oxidation suffices to functionalize the paper-based devices. The oxidized paper was applied as a sensor for protein quantification in urine, a test able to detect levels of proteinuria and even microalbuminuria. The quantification was based on the capture of proteins through covalent bonds formed with the carbonyl groups on the oxidized paper followed by the staining of the region with Ponceau dye. There is a linear dependency between human serum albumin (HSA) concentration and the length of the stained blot from 0.1 to 3 mg mL−1. This method correlated linearly with a reference method showing a higher sensitivity (0.866 cm mL mg−1) than the latter. The limit of quantification was 0.1 mg mL−1, three times lower than that of the commercial strip.
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The following files are available free of charge. FTIR spectra for all oxidized papers; ANOVA and Tukey’s analysis; variation of color intensity in the protein application spot; variation of the blot length with phosphate buffer pH; correlation of the blot length with increase of BSA concentration; analysis of HSA in urine by Bradford method and comparison between methods (PDF).
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AHI, AH, and FNC gratefully acknowledge the financial support provided by the FAPESP under the grant numbers 2017/05362-9, 2016/25806-6, and 2013/14262-7, respectively. LJMO thanks the financial support by the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior – Brasil (CAPES) – Finance Code 001. TPS gratefully acknowledges financial support from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq #154064/2016-5). EC is grateful for the INCTBio grants (FAPESP #2014/50867-3 and CNPq #465389/2014-7).
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Imamura, A.H., Segato, T.P., de Oliveira, L.J.M. et al. Monitoring cellulose oxidation for protein immobilization in paper-based low-cost biosensors. Microchim Acta 187, 272 (2020). https://doi.org/10.1007/s00604-020-04250-6
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DOI: https://doi.org/10.1007/s00604-020-04250-6