The black-footed ferret (BFF) is an induced ovulating mustelid that nearly went extinct in the 1980s. The US Fish & Wildlife Service established a captive breeding program with only seven genetic founders resulting in potential genetic drift issues that have led to reduced fecundity. Female BFFs can experience a pseudopregnancy after ovulation, characterized by similar hormone concentrations and behavior between pregnant and non-pregnant females. Our goal was to determine a noninvasive technique to identify pregnant and pseudopregnant BFFs. Our objectives were to compare fecal progesterone metabolites of pregnant (whelped, “W”) and known pseudopregnant females (identified via ultrasound after natural breeding, “DNW”, or not paired with a male and had ovulation induced using luteinizing hormone, “LH”) using three enzyme immunoassays (EIA): an inhouse pregnane (antibody CL425), inhouse pregnanediol glucuronide (PdG; antibody P26), and commercial progesterone (Arbor Assays, Inc. #K068). For pregnane, all three groups showed significant increases post-ovulation, with LH having higher pre-and post-ovulation concentrations than W and DNW. For PdG, there was no significant change in pre- and post-ovulation in all three groups, while DNW had higher overall concentrations than W and LH. For the Arbor Assays progesterone, only the W females showed a significant increase after ovulation. In summary, the commercial progesterone EIA can distinguish between pregnant and pseudopregnant BFFs; thus, providing the first steps into determining the hormonal mechanisms of pseudopregnancy while creating a tool to assist with BFF management and conservation.