Background: Pregnancy is one of the important factors affecting the quality and quantity of marital intercourses. During pregnancy, women experience physiological, psychological, and social changes as well as significant changes in hormone levels. The cultural myths that pregnant women cannot lead a normal sexual life play an important role in this respect. Aim: Evaluation of female sexual dysfunction during pregnancy and assessment of serum testosterone and estradiol with its correlation to the degree of sexual dysfunction.Patients and Methods: This study was done as a prospective case-control study involving 60 pregnant women, divided into three groups based on their trimester (1st, 2nd, or 3rd). These women attended the outpatient clinic at Obstetrics and Gynecology Department, versus 40 non-pregnant married women as controls. An Arabic validated FSFI questionnaire (ArFSFI) assessing sexual function was given to all participants and scores were calculated. Blood samples were collected to measure testosterone and estradiol levels and calculate the testosterone/estradiol ratios. Results: Arousal correlated negatively with total testosterone and estradiol in the first trimester, similar but less pronounced patterns were observed in later trimesters. The testosterone/estradiol ratio showed significant negative correlations with FSFI domains, especially in the first and third trimesters. Hormonal changes, along with female age, husband age, and duration of marriage, significantly affect sexual function during pregnancy.  Conclusion: Sexual function is significantly impaired in pregnant women compared to non-pregnant women, with notable declines in FSFI scores for desire, arousal, orgasm, and overall sexual function. The second trimester has a lesser impact on sexual function compared to the first and third trimesters. Evaluating sexual function in pregnant women should take these hormonal and demographic factors into consideration.