5α-Dihydrotestosterone (5α-DHT) is the most potent androgen in humans and its levels require regulation to maintain homeostasis. Dysregulation of 5α-DHT is associated with various pathologies, specifically excess 5α-DHT is associated with polycystic ovary syndrome (PCOS) in women and prostate cancer in men. Current treatments to manage excess 5α-DHT focus on increased expression or activity of enzymes that increase 5α-DHT biosynthesis with the goal to find inhibitors for these therapeutic targets. However, the inactivation of 5α-DHT is often ignored. Here, we find that inhibitors of 5α-DHT inactivation can contribute significantly to increase androgen receptor (AR) signaling. First, we observe in subcutaneous white adipocytes that the most prominent reaction that occurs is the conversion of 5α-DHT to its metabolite 3α-androstanediol (3α-diol), an inactivating step. To accomplish this, we developed a β-Radioactive monitoring-HPLC method to detect and quantify androgens to follow their metabolism in 5α-DHT biosynthesis and metabolism in two different white adipocyte cell lines, human Simpson-Golabi-Behmel-Syndrome (SGBS) adipocytes and primary cells isolated from a female patient. We found both SGBS and primary adipocytes favor inactivation of 5α-DHT rather than biosynthesis. Interestingly, we noted despite similar transcript levels to other inactivating enzymes, pre receptor regulator enzyme, aldo-keto reductase family 1C member 2 (AKR1C2) was the prominent pathway of inactivation. We then explore how a persistent environmental toxicant, perfluorooctanoic acid (PFOA), can disrupt AR signaling by inhibiting this very conversion. Here we show PFOA competitively and potently inhibits AKR1C2 preventing the inactivation of 5α-DHT at physiologically relevant concentrations leading to activation of AR. This provides compelling evidence for the impact of environmental contaminants on androgen signaling pathways. Furthermore, our work underscores the need for further studies into environmental influences on adipocyte steroid metabolism and their long-term effects on endocrine health.