Ambergris is a rare whale coprolith which has captivated human interest for centuries due to its unique olfactory properties and historical use in perfumery, as well in alternative medicine. The complex mixture of volatile organic compounds (VOCs) within ambergris comprises a wide range of chemical classes, including terpenes, aliphatic and aromatic hydrocarbons, and oxygenated derivatives. These compounds contribute to the distinct and alluring scent profile for which ambergris is renowned. The formation of volatile compounds in ambergris is a result of intricate biological processes within the digestive system of sperm whales, involving the interaction of ingested marine materials, enzymatic reactions, and subsequent aging and maturation of the material when deposited on beaches. The distribution and the ratios of these different compounds determine the value of the ambergris. Various methods and protocols have been engaged to assess and determine the composition. The present study focuses on the use of comprehensive two-dimensional gas chromatography coupled with time-of-flight mass spectrometry (GC×GC–TOFMS). This advanced analytical technique provides an increase in separation capacity to resolve the complexity of the ambergris volatile profile and uncover components that have not previously been reported. This novel technique being rapid, precise and convenient can bring about quick identification of authentic ambergris samples