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Smartphone-assisted visual assay platform provides novel insight for the real-time in-field quantitation of intended analytes in resource-insufficient areas. Herein, nitrogen-doped red fluorescent (FL) carbon dots (R-CDs) were developed for the timely on-site quantitation of Mn(vii) using the smartphone-assisted assay platform. R-CDs, possessing a desirable bright red FL at 616 nm under a 470 nm excitation, were fabricated through hydrothermal treatment adopting passion fruit and neutral red as precursors. Interestingly, bright red FL at 616 nm are gradually quenched upon introducing Mn(vii) based on the inner filter effect, concurrently accompanying with significant FL color variation from bright red to dark red. Inspired by the above-mentioned phenomena, hue-saturation-values (HSV) of real-time captured images could be precisely quantified through a color recognition APP within the smartphone, of which the V/S values could be employed to quantify Mn(vii) with a linear range of 50–400 μM. Furthermore, confocal fluorescence imaging of HeLa cells and zebrafish larvae demonstrates that R-CDs could be employed for the visual determination of Mn(vii) in vivoand in vitro, illustrating that R-CDs possess powerful practical application prospect in biosystem.