Homosexual women are, on average, more masculine in their appearance and behavior than heterosexual women. We hypothesized that their masculinity was influenced by exposure to elevated levels of prenatal androgen during early development. We recruited 199 women (including 67 homosexual women) and measured their masculinity via self-report and observer ratings. Our measure of prenatal androgen exposure was the ratio of the index to ring finger (2D:4D), which is hypothesized to be lower in women exposed to elevated levels of androgens during prenatal development. Homosexual women were substantially more masculine than heterosexual women in both self-report and observer ratings. However, homosexual women neither had more male-typical finger length ratios, nor did their finger length ratios relate to their masculinity in any predicted direction. Thus, we found no evidence that increased prenatal androgen exposure influenced masculinity in homosexual women.