The aim of this study was to gain knowledge of residue formation after the use of melengestrol acetate (MGA) as a growth‐promoting agent. Two Holstein‐Friesian heifers each received a daily dose through the feed of 0, 0.5 mg (2 heifers with and without withdrawal each), 1.5 mg or 5.0 mg MGA for 8 weeks. MGA residues in plasma were screened by enzyme immuno‐assay (EIA). Concentrations in kidney, liver, and muscle were quantified by liquid‐chromatography‐mass spectrometry (LC‐MS), and in fat by gas chromatography‐mass spectrometry (GC‐MS). MGA levels in plasma were 40, 128, and 280 ng/L, respectively. Residues accumulated in muscle and kidney (5‐fold), liver (20‐to‐40‐fold), and fat (200‐fold). After administration of 1.5 mg per day the mean MGA concentration in fat was 29 μg/kg and thus violated USA regulations which specify a limit of 25 ppb. Therefore the labelled use of MGA (0.5 mg per day) has to be officially controlled.