The progesterone (P(4)) concentration (ng/5 mul) in the fat portion of cow's milk was measured on days 0, 12, 20, 24, and 30 after insemination in an effort to assess the ability of this analysis to judge the reproductive status of dairy cows. Pregnancy was determined by rectal palpation at approximately 40 days after insemination. The mean P(4) concentration (ng/5 mul) +/- SD in the pregnant cows (n=17) was 0.14+/-.07, 1.31+/-.36, 1.41+/-.52, 1.22+/-.21, and 1.30+/-.43 on post-insemination days 0, 12, 20, 24, and 30 respectively. At these same intervals the P(4) concentration in the non-pregnant (NP) cows (n=18) was 0.14+/-.06, 1.26+/-.37, 0.56+/-.52, 0.57+/-.42, and 1.08+/-.59. On days 20 and 24 after insemination, mean P(4) levels were significantly (P<.001) elevated in the pregnant cows. It was concluded that an accurate assessment of the reproductive status of a cow, milk samples from days 0, 20, and 24 post-insemination would have to be analyzed for milkfat P(4) concentrations. In order to determine the percentage of cows inseminated out of the periestrual period, milkfat P(4) concentrations were ascertained in milk collected from 165 cows at the time of insemination. Cows that conceived (n=38) had a mean milkfat P(4) concentration at the time of insemination of 0.16+/-.09. The upper limit for P(4) in the milkfat at the time of insemination in cows that conceived was calculated to be 0.43 ng/5 mul milkfat. Subsequently, it was found that of the 14 cows that had P(4) levels above this upper limit at time of insemination, nine were inseminated at a period other than in the periestrual period and five were inseminated while already pregnant.