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Spheres, tubes, and planar-shaped nanomaterials as Fe3O4 nanoparticles (MNPs), multiwalled carbon nanotubes (MWCNT), and graphene oxide (GO) were used for the first time to treat microfluidic paper-based analytical devices (μPADs) and create a biocompatible layer with high catalytic surface. Once glucose measurements are critical for diabetes or glycosuria detection and monitoring, the analytical performance of the proposed devices was studied by using bienzymatic colorimetric detection of this carbohydrate. The limit of detection values achieved for glucose with μPADs treated with MNPs, MWCNT, and GO were 43, 62, and 18 μM, respectively. The paper surface modification solves problems associated with the lack of homogeneity on color measurements that compromise the sensitivity and detectability levels in clinical diagnosis.