Palatal rugae are anatomical ridges on the anterior palatal region, noted for their uniqueness and stability, which makes them valuable in forensic identification. This study investigated gender-based differences in palatal rugae patterns among adults aged 20–40 years and evaluated their potential role in sex determination. A total of 20 participants (10 males and 10 females) were included. Maxillary impressions were taken, and dental casts were prepared using dental stone. The palatal rugae were measured bilaterally for their number and length using a Vernier caliper and classified into primary, secondary, and fragmentary types. Data were statistically analyzed using paired and independent Student's t-tests. No significant differences were found in the number or length of rugae between the right and left sides of the palate (p > 0.05). A gender-wise comparison revealed no significant difference in the total number of rugae (p = 0.89) or in the length of primary rugae (p = 0.90). However, females exhibited significantly longer secondary rugae (p = 0.02), whereas males had a significantly higher number of fragmentary rugae (p = 0.04). Palatal rugae display gender-specific differences in secondary and fragmentary patterns and may serve as supplementary indicators in forensic sex determination. Further studies with larger sample sizes are recommended to substantiate these findings.