Background: Premature rupture of membranes (PROM) is a significant cause of perinatal morbidity and mortality, necessitating early diagnosis and intervention. Current diagnostic methods for PROM have limitations, highlighting the need for a reliable biomarker. Creatinine, present in vaginal secretions, may indicate PROM due to its ability to pass from the maternal bloodstream to the amniotic fluid. This study aimed to investigate the correlation between creatinine levels in vaginal secretions and PROM, contributing to the development of a non-invasive diagnostic approach and improved management strategies for better outcomes. Methods: A prospective observational study was conducted over two years, enrolling 129 pregnant women with premature rupture of membranes (PROM) in the third trimester (Group A) and establishing a control group (Group B) of pregnant women without complications. Sample size was calculated to ensure statistical significance. Vaginal secretions were collected using sterile techniques, and creatinine levels were quantitatively analyzed using a biochemical assay. Statistical analysis, including descriptive statistics and appropriate tests, was performed using SPSS version 20.0. Ethical considerations were upheld, ensuring participant privacy and confidentiality. Results: The mean age of participants in present study was 27.38 ± 4.12 years in Group A (with PROM) and 28.24 ± 4.28 years in Group B (without PROM).In present study, there were 57 primiparous women, accounting for 44.2% of Group A (with PROM), and 52 primiparous women, making up 39.1% of Group B (without PROM).In present study, the mean gestational age in weeks for Group A (with PROM) was 32.20 ± 3.24 weeks, while for Group B (without PROM) it was 32.67 ± 3.48 weeks.In present study, the mean creatinine levels in mg/dL for Group A (with PROM) were 1.89 ± 0.63 mg/dL, while for Group B (without PROM) they were 0.57 ± 0.25 mg/dL.In our study, the area under the curve (AUC) was 0.906 (95% CI: 0.866-0.943), indicating excellent diagnostic accuracy of creatinine in vaginal wash to diagnose PROM. Conclusion: In conclusion, the present study provides valuable insights into the association between various variables, including age, parity, gestational age, foetal presentation, cervical dilation, and biomarkers such as creatinine and amniotic fluid index, with the occurrence of PROM.