Case summary A 9-year-old male neutered European Shorthair cat was presented owing to vomiting and mild weight loss. Clinical examination was normal, but biochemistry results showed increased concentrations of total calcium (4.05 mmol/l; reference interval [RI] 2.20–2.90 mmol/l) and ionised calcium (iCa) (2.19 mmol/l; RI 1.12–1.40 mmol/l), as well as hypophosphataemia (2.5 mg/dl; RI 3.1–7.5 mg/dl). Parathyroid hormone (PTH) concentration (>1000 pg/ml) was markedly increased, while parathyroid hormone-related protein concentration (<0.8 pmol/l) was normal. Neck ultrasound showed a large left parathyroid mass (13 × 7 × 6 mm). Under general anaesthesia and with ultrasonographic guidance, a fine-needle aspiration of the mass followed by chemical ablation with 2 ml 96% ethanol was performed. The cat was re-evaluated and iCa concentration measured 24 h, 72 h, 5 days, 4 weeks and 4 months post-ablation. Normocalcaemia was reached within 24 h, remained stable throughout the whole evaluation period and the concentration of PTH normalised 4 months later. Vomiting stopped promptly after chemical ablation and a slight change in voice, as well as a mild prolapse of the nictitating membrane, were the only side effects after the treatment but resolved some weeks later. Relevance and novel information To our knowledge, this is the first report of successful chemical ablation of a parathyroid mass in a cat with primary hyperparathyroidism. Chemical ablation might therefore be a possible alternative to parathyroidectomy in cats.