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Equine urine was analysed by capillary gas chromatography. The volatile profiles from oestrous and dioestrous samples were compared to establish any qualitative or quantitative difference that may have potential value in olfactory communication. Forty-five different volatile compounds were detected. Of these, 17 major compounds were common to all chromatograms. The chemical profile of oestrous urine was distinguished by the presence of a unique peak that was not present in dioestrous samples. Numerous constituents exhibited endocrine dependence: while the concentrations of seix peaks increased at oestrus, the concentrations of another five peaks decreased at the same time. Since oestrous urine, but not dioestrous urine, has been shown to elicit sexual behaviour in the stallion, the unique peak, together with the peaks that were present in increased concentrations at oestrus, may represent important chemical signals that stallions use to detect urinary ‘oestrous odours’. Statistical analyses also indicated that the relative ratios (normalized peak areas) of many peaks changed significantly across the oestrous cycle: the ratios increased in nine peaks, decreased in six peaks, and remained constant in two peaks at oestrus.