Benign prostatic hyperplasia (BPH) is a prevalent condition among older men, characterized by prostate gland enlargement due to the proliferation of stromal and epithelial cells. It is the fourth most commonly diagnosed disorder in elderly men and the leading cause of lower urinary tract symptoms in this population. Histologically identifiable BPH affects over 50% of men by age 60 and nearly 90% by age 85. The condition is closely associated with dihydrotestosterone (DHT), a potent androgen produced from testosterone by the enzyme 5α-reductase, which promotes prostate growth. While the complete pathogenesis of BPH remains partially understood, androgens are known to play a critical role, and inhibiting DHT production or action has proven effective in reducing prostate growth. In this study, in vitro experiments were conducted to evaluate the 5α-reductase inhibitory potential of extracts from Carthamus tinctorius and Carthamus oxycantha. Enzyme solutions were prepared through prostate homogenization, with enzyme concentration determined using Bradford reagent. The inhibitory activity was assessed using a standard testosterone curve, with peak areas measured by HPLC at 254 nm, employing a methanol: water (70:30) mobile phase and a flow rate of 1 ml/min. Optimum enzyme concentration was established by varying enzyme levels while keeping substrate concentrations constant. The results revealed that the n- hexane fraction of the methanolic extract of Carthamus tinctorius exhibited significant 5α-reductase inhibitory activity, with an IC50 value of 0.301 mg. In contrast, chloroform, methanol, butanolic fractions, and various other extracts did not demonstrate significant inhibitory potential. These findings suggest that Carthamus tinctorius, particularly its hexane fraction, holds promise for therapeutic applications in managing prostatic hyperplasia, while highlighting the differential pharmacological properties of Carthamus oxycantha.