p- and m-Cresols emitted from estrous urine are reliable volatile chemical markers of ovulation in mares

Urine samples from 14 mares, belonging to five breeds, were collected at estrus and diestrus to search for estrous specific volatile compounds which could be used to determine ovulation time. Around 150 volatiles were collected from urine head-space samples by solid phase micro extraction technique, and analyses were conducted by gas chromatography-mass spectrometry methods. Comparison of chromatographic profiles of volatile substances revealed that concentrations of m- and p-cresols were significantly greater during estrus while diethylphthalate was more abundant at diestrus. Monitoring of m- and p-cresols during the period of estrus and a few days before and after estrus revealed irregular changes in amounts of cresols until 3-4 days before ovulation when the concentration of the compounds began to increase with peaks 1 day before ovulation. On the day when ovulation occurred, amounts of the metabolites decreased sharply, almost to basal concentrations, and remained at these concentrations for 6 days - when sampling was finished. In four of the mares changes in the concentration of diethylphthalate were less pronounced and more temporally variable compared with those of cresols. Based on reproducible temporal changes in concentrations of m- and p-cresols, with respect to the time of ovulation, a noninvasive test to determine a precise insemination time could occur. This would save time, reduce costs and simplify the procedure.