A hematocrit (Hct) of less than 25% during cardiopulmonary bypass (CPB) and transfusion of homologous packed red blood cells (PRBC) are each associated with an increased probability of adverse events in cardiac surgery. Although the CPB circuit is a major contributor to hemodilution intravenous (IV) fluid volume may also significantly influence the level of hemodilution. The objective of this study was to explore the influence of asanguinous IV fluid volume on CPB Hct and intraoperative PRBC transfusion. After Institutional Review Board approval, a retrospective chart review of 90 adult patients that had undergone an elective, isolated CABG with CPB was conducted. Regression analysis was used to determine if pre-CPB fluid volume was associated with the lowest CPB Hct and the incidence of an intraoperative PRBC transfusion. In separate multivariate analyses, higher pre-CPB fluid volume was associated with lower minimum CPB Hct (p < .0001), and higher minimum CPB Hct was associated with a decreased probability of PRBC transfusion (p < .0001). Compared to patients that received <1600 mL (n = 55) of pre-CPB fluid, those that received >1600 mL (n = 35) had a decreased mean low CPB Hct (22.4% vs 25.6%, p < .0001), an increased incidence of a CPB Hct <25% (74% vs. 38%, p = .0008) and PRBC transfusion (60% vs. 16%, p < .0001), and increased median PRBC units transfused (2.0 vs 1.0, p = .1446) despite no significant difference in gender, age, patient size, baseline Hct, or CPB prime volume. Patients that received a PRBC transfusion (n = 30) received a significantly higher volume of pre-CPB fluid than nontransfused patients (1800 vs. 1350 mL, p = .0039). These findings suggest that pre-CPB fluid volume can significantly contribute to hemodilutional anemia in cardiac surgery. Optimizing pre-CPB volume may preserve baseline Hct and help limit intraoperative hemodilution.