Glucose is an analyte of great importance, both in the clinical and sports fields. Since blood is the gold standard biofluid used for the analytical determination of glucose, there is high interest in finding new non-invasive biofluids, such as sweat, for its determination. This research aims at developing a novel and biocompatible alginate bead-like biosensor for glucose sensing in artificial sweat, as well as at the fabrication of a microfluidic device for the integration of the biosensor, capable of performing sequential analysis of glucose at different times, this is, chrono-sampling. The detection of glucose was achieved through an enzymatic reaction and the image colour analysis of the product. Glucose in artificial sweat was detected both in bead and in device in the physiological range of 10 – 1000 μM. Moreover, the biosensor, which contained titanium nanotubes to increase the hydrophilic character of the bead, was successfully integrated in a polydimethylsiloxane device, allowing chrono-sampling over 42 min. This novel sensing material and the device are oriented towards the development of a miniaturized device for fast analysis at the point and time of need. Moreover, the possibility of chrono-sampling opens the possibility for the continuous monitoring of sweat biomarkers during exercise, allowing a personalized- oriented training to improve athletes’ performance.