Prolonged occupational exposure to waste anesthetic gases may have the potential to cause adverse health effects. Workplace exposure surveillance programs are intended to reduce health risk by evaluating exposures to waste anesthetic gases during surgical procedures. Both the personal breathing-zone and area measurements are used to assess occupational exposure in the operating theater. Direct-reading instruments provide real-time measurements and are useful for identifying leaks and evaluating on-the-spot corrective actions. Passive diffusion monitors quantify occupational exposures over time during surgery. The aim of this study was to evaluate a waste anesthetic gas surveillance program to understand occupational exposures and further improve data collection strategy. For this study, 76 survey reports from 2012 through 2014 were retrospectively reviewed to assess occupational exposures to isoflurane in 58 unique procedural rooms operated by the National Institutes of Health. The surveys included industrial hygiene assessments performed during animal and human surgical procedures. The survey reports were evaluated qualitatively and data from these reports was transcribed for quantitative analysis. Variations in sample strategy were observed between surveys and were attributed to ambiguity in the written surveillance program. The study also evaluated the relationship between isoflurane concentrations and sampling method, sampling location, patient type, or scavenging method. Isoflurane exposures were significantly higher among procedures performed on rodents compared to the patients with a large body mass (humans, non-human primates, and swine) (P < 0.05) and in procedures using the charcoal canister exhaust system compared with the central vacuum exhaust system. In addition, individuals performing the surgical procedure experienced elevated occupational exposures measured by both direct-reading instrument and passive diffusion monitors, that is, exposure was significantly higher as measured at the breathing-zone compared with any area within the room (P < 0.05). The study identified several inconsistencies and shortcomings in the surveillance program. Isoflurane concentrations measured during rodent procedures requires further review of work practices and engineering controls. Overall, the findings provide insights to further improve data collection, monitoring, and control of isoflurane exposures.