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• A smartphone-interfaced flow injection sensing system was developed. • An electrode was modified with Pd-PANI/Carbon microspheres@carbon nanotubes. • Sensitive determination of glucose was achieved in real human serum samples. Integrating a flow injection system with an electrochemical sensor controlled by a smartphone enables rapid, precise real-time monitoring and reduces analyte consumption, contamination and costs. An enzyme-free electrochemical sensor is presented for the monitoring of glucose, an important indicator of diabetes. The sensor integrates a smartphone and a flow injection amperometric system. The flow cell is fabricated from acrylic plastic and includes an electrochemical three-electrode cell that comprises a working electrode modified with a catalyst of palladium-polyaniline/carbon microspheres@carbon nanotubes (Pd-PANI/CM@CNTs/E) for the detection of glucose oxidation. In the optimal condition, the proposed system produced good analytical performances. The sensor determined glucose linearly in a range from 0.020 to 100 mM, with a sensitivity of 51.51 μA mM−1 cm−2, and a detection limit (LOD) of 0.0071 mM. The developed system also exhibited good repeatability, reproducibility, operational stability, and anti-interference properties, and accurately determined glucose levels in real human serum samples. This sensing platform provides an alternative tool for the development of a continuous glucose monitor system for clinical applications. [Display omitted] [ABSTRACT FROM AUTHOR]