Accuracy and precision of a prototype point-of-care (POC) hemoglobin (Hb) and oxygen saturation (SO2) analyzer were compared to a benchtop analyzer, as well as the use of the prototype in a field setting. Arterial and venous Hb concentrations ([Hb]) and SO2 were determined from 180 whole-blood samples from dogs, cats, and horses. Hemoglobin concentrations and SO2 values were consistently lower (P < .0001) for the prototype compared to the benchtop analyzer. Deming's regression and Bland–Altman bias representation with concordance analysis revealed good accuracy and precision but poor concordance. When separated out by species, concordance was moderate to excellent for canine but poor for equine samples; accuracy and precision were unchanged. When separated by sample type, there was loss of accuracy and precision for equine arterial SO2 and canine venous SO2. Whole-blood jugular venous [Hb] and SO2 values determined for 26 horses before and after exercise using the prototype analyzer in a wide range of temperatures revealed good consistency and precision. In conclusion, the prototype POC had good concordance with the benchtop analyzer for canine but not equine samples, with good accuracy and precision for equine and canine [Hb] and SO2. Concordance results indicate the prototype's calibration settings may need to be adjusted for different species and sample type if using the comparative benchtop analyzer's reference values. Overall, the prototype POC analyzer had good accuracy and precision for the two analytes for both species and sample types, was simple and practical to use in the field, and may be a suitable substitute for a benchtop analyzer.