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Aim: To describe lipid oxidation during physical activity (PA) in overweight and obese patients using a real-time capillary glycerol sensor and to propose a personalized strategy to optimize lipolysis., Methods: Healthy adult volunteers with a BMI >25 kg/m2 were recruited. All participants performed 27 sessions of 30-min PA covering all possible combinations of exercises (low, moderate, high intensities) and pre-exercise meals (high-carbohydrate, high-fat, or fasting) with 3 replicates. Glycerol measurements were performed five times during each session: at T0, T20, T30, T45, and T60. The evolution of capillary glycerolemia during PA was modeled by a nonlinear mixed-effects model. We developed a personalized PA recommendation based on the first 2 glycerolemia measurements that indicates whether a change in activity type is preferable., Results: Thirteen patients were included in the study and 1232 capillary glycerolemia measurements were performed. Hyperglycemic and hyperlipidemic meals were associated with lower baseline glycerolemia (p < 0.0001 for both) compared to fasting. In contrast, the type of meal was not significantly associated with PA-induced lipolysis (p = 0.52 and p = 0.76 for high-carbohydrate and hyperlipidemic meal respectively). Compared with exercises done at low and moderate intensities, high intensity exercises were associated with a higher PA-induced lipolysis (p = 0.0002). Specific strength exercises were associated with a lower PA-induced lipolysis (p < 0.0001). The 2 initial glycerolemia measurements appeared to be a pertinent criterion which help the personalization of PA programs for such individuals (c-index 0.67)., Conclusion: The inter-individual lipolysis responses to PA and the availability of a real-time capillary blood glycerol sensor may enable the personalization of PA programs, thus promoting optimal lipolysis in subjects with overweight and obesity., Competing Interests: Declaration of competing interest All co-authors declare no relationship of interest with the study presented in this article., (Copyright © 2024 The Author(s). Published by Elsevier Ltd.. All rights reserved.)