Abstract Purpose: In this study, the Pre-implantation embryonic sex ratio in 125 couples who had three or more female children and underwent pre-implantation genetic diagnosis (PGD) for non-medical reason was included. Besides, we have aimed to find out whether these couples had more chances of getting a girl child once again. Methods: 125 couples who had three or more female offspring and those who underwent PGD for non-medical sex selection (XY) between 2015 and 2019 were included. Nuclear DNA was analyzed by Fluorescent in situ Hybridization (FISH). 2-chromosome (X, Y), 3-chromosome (21, X, Y), 5-chromosome (13, 18, 21, X, Y) probes were used for FISH. The standard protocol was followed for sperm processing and embryo culture for IVF and PGD. Result: Independent sample t-test showed that there is no significant difference between equal and unequal embryonic groups in patients’ age, husbands’ age, sperm count, sperm motility, total male embryos, total female embryos, normal male embryos, and normal female embryos. For patients with positive pregnancy outcome, 84.6% had unequal embryonic ratio while 15.4% had equal embryonic ratio. Similarly, patients those who were treated by short protocol had 85% of unequal embryonic ratio and 15% had equal ratio. Conclusion: Greater variability in the female to male embryos was found in these couples, confirms the fact that couples previously having girl offspring may predominantly not produce embryo of the same sex every time.