Volcanic glass has been used extensively as a paleoaltimeter. Deuterium (2H) concentrations in glass have been found to be stable over geologic timescales, making delta 2H (also known as delta D) a reliable proxy for ancient water chemistry. However, continued work revolves around better understanding how different factors affect preserved water in volcanic ash. Here, we analyze delta D in the Rattlesnake Tuff (RST), a widespread ca. 7 Ma ash-flow tuff, and create a paleoisoscape to assess variations in delta D across Oregon during that time. To this end, 16 ash samples were collected across central and eastern Oregon from various flow units within the RST. Samples were analyzed for delta D using a temperature conversion elemental analyzer (TC/EA) connected to a mass spectrometer and elemental composition using a scanning electron microscope (SEM). We compared the isotopic results to modern water and published ancient water proxy data to better constrain changes in climate and topography across Oregon throughout the Neogene. We also estimated wt. % H2O by calculating excess (non-stoichiometric) oxygen from SEM elemental data. We did not observe significant variations in delta D among the flow units from single locations, nor was there a significant relationship between the prepared glass shard composition and wt. % H2O or delta D, supporting the use of volcanic glass as a reliable paleoenvironmental indicator. Our results show significant spatial variation in delta Dwater values of RST, ranging from -107 parts per thousand to -154 parts per thousand. delta D values of ancient glass were similar to modern water near the Cascade Mountains but became relatively negative to the east near the inferred eruptive center of the RST, suggesting that a significant topographic feature existed in the vicinity of the RST eruptive center that has since subsided.