Chloralose (alpha-chloralose) is a poisonous substance currently used as a rodenticide or avicide. It has primarily been used in Europe since 1893 as a human and veterinary hypnotic agent. Chloralose is a central nervous system depressant also acting as a stimulant on spinal reflexes. In the present case, a 24-year-old man was found dead in his bedroom near vomit residues. Several items were seized from the scene, including an empty bottle of Murex 50 g (α-chloralose), sold in Italy as rodenticide. Postmortem examination revealed no evidence of natural disease or trauma. Heart blood, urine, gastric contents, vitreous humour, brain, bile and liver were collected and submitted for toxicological analysis. Several extraction procedures and a specific liquid chromatography-tandem mass spectrometry protocol were purposely developed and validated. Chloralose was found in blood at a concentration of 65.1 mg/L and high levels were also detected in the gastric contents, confirming its ingestion shortly before the man's death. The distribution of chloralose in the body was evaluated by analyzing urine, vitreous humour, brain, bile and liver specimens. Quantitation of chloralose in several body fluids and tissues adds new data about the distribution of this chemical in the human body after massive ingestion.