Conventional approaches of training ecohydrologic models usually consider one hydrometric dataset, such as field measured matric potential or a soil moisture time series. In numerical modeling, these metrics can be used to optimize soil physical parameters using equations for water content and hydraulic conductivity. However, training a model on matric potential alone neglects valuable calibration potential available from transport processes, often described by the advection dispersion equation. Our group aims to quantify this potential by using stable isotopes of water as a tracer. In HYDRUS-1D, we couple daily stable isotope ratios and matric potential measurements to optimize soil hydraulic and transport parameters for two forested sites in the Upper Colorado River Basin. Using inputs of δ2H, five parameter optimization schemes were tested across four sampling intervals (daily, weekly, biweekly, and rainfall-based) to assess calibration performance. Results show that when KGEΨ values are highest (indicating strong model performance with regard to matric potential), unincorporated values of KGED are considerably low (indicating poor model performance with regard to stable isotopes). We hypothesize that the maximum average of both metrics, KGEx̄, yields a more robust parameter set, and consequently, a more physically representative model. Comparing KGEx̄ for daily and weekly sampling intervals shows a negligible difference, suggesting that weekly sampling is adequate for model calibration. Rainfall-based sampling (two days after precipitation ≥ 3 mm) shows promise, with a comparable model performance to daily and weekly sampling. Meanwhile, the model underperformed with biweekly sampling. We propose that weekly sampling is sufficient for producing an optimal parameter set, and that daily sampling is likely unnecessary. For good measure, less frequent sampling intervals are not recommended. This study aims to evaluate sampling frequency and model performance to benefit future efforts of ecohydrologic model calibration.