The impact of rising temperature variability driven by climate change on metabolic health remains understudied, especially considering the global increase in diabetes prevalence, with long-term effects on glucose metabolism unexplored. This study investigated associations between long-term temperature variability exposure and glucose metabolism in a population-based cohort of 2997 participants (4954 observations) over a 7-year period from KORA F4 and FF4 cohorts in Augsburg, Germany. Long-term exposure to temperature variability was estimated as the standard deviation of the daily mean air temperature over the 365-day period preceding each examination. We applied generalized estimating equations to examine the longitudinal associations between long-term exposure to temperature variability and multiple glucose metabolism biomarkers: fasting glucose, 2h glucose, fasting insulin, homeostasis model assessment of insulin resistance (HOMA-IR), homeostasis model assessment of β-cell function (HOMA-B), quantitative insulin sensitivity check index (QUICKI), and glycated hemoglobin (HbA1c). We found that a 1 °C higher temperature variability was significantly associated with higher fasting insulin, HOMA-IR, and HbA1c with % changes (95% CI) of 2.62 (0.79; 4.49), 2.81 (0.79; 4.87), and 2.38 (1.97; 2.79), respectively, and lower QUICKI (-0.41 [-0.70; -0.11]). These findings suggest that increasing temperature variability exposure may contribute to metabolic dysfunction, potentially accelerating the global diabetes epidemic.