Diabetes has surged dramatically, leading to a significant healthcare burden worldwide. Consequently, there is a growing need for reliable non-invasive alternatives for glucose monitoring, replacing traditional blood-based analysis. Despite this, enzyme degradation and stability have posed challenges for sweat sensors. To address these obstacles, the present study emphasizes the development of a novel, non-enzymatic electrochemical sweat sensor for glucose monitoring that is painless, reliable, and based on polyaniline nanocaps. Polyaniline (PANI) nanostructures, including nanocaps, nanowires, and nanocollides, were successfully synthesized using sodium dodecyl sulphate (SDS) as a template at varying ratios. The obtained nanostructures were characterized using XRD, FTIR, FESEM, and TEM. For electrochemical analysis, a screen-printed carbon electrode (SPCE) was employed. The presence of subsistent polyaniline nanocaps structure resulted in better catalytic performance, exhibiting superior sensitivity of 94.3859 µA mM−1 cm−2 and a limit of detection (LOD) of 0.04 µM in the linear range of 10–500 µM for glucose monitoring. Furthermore, the stability of the modified PANI nanocaps@SPCE sensor was tested using a mimic sweat sample. These results suggest that the fabricated PANI nanocaps@SPCE sensor holds promise as a non-invasive sweat sensor for glucose monitoring.