Selected blood chemistry and gas reference ranges for clinically healthy broiler breeder hens were established using CG8 cartridges in an i-STAT® handheld point-of-care clinical analyzer. Samples from 165 hens (25–36 wk of age), representing three broiler breeder strains reared by four integrators, were evaluated. A standardized sampling technique was developed to minimize instrument error readings. The following reference ranges and means, respectively, were determined: sodium (141.6–152.6, 147.1 [mmol/L]), potassium (4.1–5.7, 4.9 [mmol/L]), ionized calcium (1.20–1.73, 1.47 [mmol/L]), glucose (207.2–260.7, 234.0 [mg/dl]), hematocrit (21.3–30.8, 26.1 [% packed cell volume]), hemoglobin (7.3–10.5, 8.9 [g/dl]), pH (7.28–7.57, 7.42), carbon dioxide partial pressure (25.9–49.5, 37.7 [mm Hg]), oxygen partial pressure (32.0–60.5, 46.2 [mm Hg]), bicarbonate (18.9–30.3, 24.6 [mmol/L]), total carbon dioxide (19.9–31.5, 25.7 [mmol/L]), base excess (−6.8 to 7.2, 0.2), and oxygen saturation (70.6–93.3, 82.0 [%]). Wide ranges in blood gases and base excess occurred in all strains. Cobb strain hens had significantly lower glucose and higher partial and saturated oxygen values compared with two Ross strains. Significant differences in several blood parameters were found among different integrators and in older postpeak production birds. The i-STAT handheld point-of-care clinical analyzer provides rapid, relatively low cost, blood chemistry values that are useful for investigating broiler breeder flock diseases of unknown or uncertain etiology, especially those suspected of having a metabolic cause.