Partitioning of evapotranspiration (ET) in urban forest lands plays a vital role in mitigating ambient temperature and evaluating the effects of urbanization on the urban hydrological cycle. While ET partitioning has been extensively studied in diverse natural ecosystems, there remains a significant paucity of research on urban ecosystems. The flux variance similarity (FVS) theory is used to partition urban forest ET into soil evaporation (E) and vegetation transpiration (T). This involves measurements from eddy covariance of water vapor and carbon dioxide fluxes, along with an estimated leaf-level water use efficiency (WUE) algorithm. The study compares five WUE algorithms in partitioning the average transpiration fraction (T/ET) and validates the results using two years of oxygen isotope observations. Although all five FVS-based WUE algorithms effectively capture the dynamic changes in hourly scale T and E across the four seasons, the algorithm that assumes a constant ratio of intercellular CO2 concentration (ci) to ambient CO2 concentration (ca) provides the most accurate simulation results for the ratio of T/ET. The performance metrics for this specific algorithm include the RMSE of 0.06, R2 of 0.88, the bias of 0.02, and MAPE of 8.9 %, respectively. Comparing urban forests to natural forests, the T/ET in urban areas is approximately 2.4–25.3 % higher, possibly due to the elevated air temperature (Ta), greater leaf area index (LAI), and increased soil water availability. Correlation analysis reveals that the T/ET dynamic is primarily controlled by Ta, LAI, net radiation, ca, and soil water content at half-hourly, daily, and monthly scales. This research provides valuable insights into the performance and applicability of various WUE algorithms in urban forests, contributing significantly to understanding the impact of urbanization on energy, water, and carbon cycles within ecosystems.