Ketosis is a metabolic disorder associated with high production and low energy intake in dairy cows. Fat reserves are mobilized, and gluconeogenesis occurs. Traditionally, rapid tests for blood, milk or urine are used to detect increased ketone body levels in case of ketosis. Breath analysis is relatively new and relevant for the development of automatic sensor systems for early warning. This study aims to determine whether and when a postpartum rise in ketone bodies occurs in breath with elevated blood β-hydroxybutyrate (BHB) in cows at risk of ketosis. Postpartum breath, blood, urine and milk samples were taken daily until day 7, with one prepartum sample excluding milk, and ketone body concentrations were determined. Concentrations of blood BHB were 0.4–2.6 mmol/L (lab) and 0.3–3.1 mmol/L (rapid test), breath acetone was 2.3–20.0 ppm, urine acetoacetate 0–8 mmol/L and milk BHB 0–500 µmol/L. A rise in blood BHB was related to that in urine acetoacetate and milk BHB and followed by a rise in breath acetone. However, breath acetone levels of ketotic and non-ketotic cows were similar. We conclude that rising ketone bodies can be detected in blood, urine, milk and breath, but to use breath analysis as an alternative to rapid tests, longitudinal sampling is needed.