The goal of this study is to measure and compare the hormonal serum levels (estrogen, progesterone, testosterone, prolactin, dihydrotestosterone [DHT]) in trigeminal neuroglia (TN) menopausal women and healthy women. This cross-sectional and case-control study was performed in 2018 and 2019. For this study, menopausal women with confirmed TN were enrolled. Twenty-two healthy women in the control group and 19in the case group participated. Blood samples weretaken from participants for assessment of hormonal serum levels (estrogen, progesterone, testosterone, prolactin, DHT). Data were analyzed by SPSS version 18. Mann-Whitney, T-test, kormography test, nonmetric, χ2 test, and odds ratios have been used. In patients with TN, the serum level of testosterone was significantly higher (p = .036), and the serum level of prolactin (p = .016) was significantly lower. Other evaluated hormones'serum level was identical in the two groups. Patients with abnormal estrogen levels were more in the TN group in comparison with the healthy group. The abnormality of progesterone in TN patients was more in comparison to the healthy control group. Estrogen and progesterone serum levels in TN patients are higher in comparison with the healthy group, while prolactin and testosterone serum levels are lower in the control group. Moreover, the DHE serum level is similar in both groups.