Cryogenic vacuum distillation (CVD) is a widely used technique for extracting plant water from stems for isotopic analysis, but concerns about potential isotopic biases have emerged. Here, we leverage the Cavitron centrifugation technique to extract xylem water and compare its isotopic signature to that of CVD-extracted stem water as well as source water. Conducted under field conditions in tropical northern Australia, our study spans seven tree species naturally experiencing a range of water stress levels. Our findings reveal a significant deuterium bias in CVD-extracted bulk stem water when compared to xylem water (median bias \($-14.9\textperthousand$\)), whereas xylem water closely aligned with source water (median offset \($-1.9\textperthousand$\)). We find substantial variations in deuterium bias among the seven tree species (bias ranging from -19.3 to \($-9.1\textperthousand$\)), but intriguingly, CVD-induced biases were unrelated to environmental factors such as relative stem water content and pre-dawn leaf water potential. These results imply that inter-specific differences may be driven by anatomical traits rather than tree hydraulic functioning. Additionally, our data highlight the potential to use a site-specific deuterium offset, based on the isotopic signature of local source water, for correcting CVD-induced biases.