Background The end of pregnancy is the period with the highest risk of occurrences of ketosis and pregnancy toxemia due to fat mobilization and increasing non-esterified fatty acids in the liver which are converted in ketone bodies, mainly β-hydroxybutyrate acid (BHB). This ketone body may also become elevated in the bloodstream. The present study validates the use of a handheld meter for determining the blood concentration of BHB and ascertaining the predictive value and accuracy of BHB measurements in diagnosing hyperketonaemia in ewes. Methods A total of 19, non-pregnant, crossbred ewes were subjected to 2 h of intravenous infusion of a saturated BHB solution. Over 6 h of evaluation, 247 blood samples were obtained in 13 sampling moments. The BHB concentration was measured by an enzymatic colorimetric method in an automated biochemical analyzer (gold-standard) and by a handheld meter using an electrochemical enzyme technique. Results There was a high correlation between both methods (r = 0.98; P < 0.001). Considering the blood BHB concentrations range 0.8–1.6 mmol/L for moderate ketosis the handheld meter presented sensitivity and specificity of 0.98 and 0.81, respectively. For severe ketosis (BHB ≥ 1.6 mmol/L) sensitivity and specificity were 0.99 and 0.75, respectively. Thus, the handheld device can be useful for diagnoses of cases of mild or severe pregnancy toxemia at field conditions.