False claim/labels on commercial products with high-sugar can lead to serious health issues; therefore, fast analytical method for qualitative and quantitative detection of sucrose/table sugar is very important. Rapid and sensitive electrochemical sensors based on composite hydrogel matrix of microfibrillated cellulose (MFC), Indian gum Tragacanth (IGT) with reinforced nanomaterials and co-entrapped enzymes were prepared for sucrose analysis in tea samples. Gold screen printed electrodes (AuSPEs) were modified with multilayered nanocomposite of MFC-IGT as continuous matrix with invertase (INV)-glucose oxidase (GOx) and gold/copper nanoparticles (AuNPs/CuNPs) as filler and their electrochemistry was studied. Probe A (INVWM-GOx-AuNPs-MFC-IGT/AuSPE) demonstrated the sucrose detection range from 10 mu M to 10 mu M while Probe B (INVWM-GOx-CuNPs-MFC-IGT/AuSPE) and Probe C (INVWM-GOx-Au/CuNPs-MFC-IGT/AuSPE) displayed a wider linear range (0.1 nM to 10 mu M) and better limit of detection. Based on performance, probes B and C were used to detect sucrose content in sugar-sweetened beverages i.e. tea samples. The novel nanocomposite polymeric-backbone of MFC-IGT proved its highest sensitivity towards sucrose. All modified-gold probes exhibited higher reproducibility as well as better long-term stability and reusability with fast response time (<5 s) as compared to previous reports. Thus, the proposed methodology can be promising tool in food and biomedical industry with slight modifications. (C) 2019 The Electrochemical Society.