OBJECTIVE Obesity is characterized by dysregulated adipokine production patterns, leading to low-grade inflammation. Irisin has been identified as a potential target for the treatment of diabetes and obesity. Nesfatin-1, another adipokine, plays a significant role in various metabolic processes, including glucose homeostasis. This study investigated the levels of the new generation adipokines, irisin and nesfatin-1, in the amniotic fluid during the second trimester of normal and obese pregnancies. METHODS Amniotic fluid samples were collected following established protocols. The first 2 mL portion of fluid obtained during amniocentesis was retained aAfter centrifugation at 1500 g for 10 minutes, the cell-free amniotic fluid was transferred to Eppendorf tubes and quantitative measurements of irisin and nesfatin-1 levels were performed using the Enzyme-Linked Immuno Sorbent Assay (ELISA) method. RESULTS Our results demonstrate, for the first time in the literature, the presence of irisin and nesfatin-1 in amniotic fluid. Additionally, we found that the levels of these adipokines were significantly lower in obese pregnant women compared to the control group (both p<0.05). CONCLUSION By establishing the presence of these hormones in amniotic fluid, our study provides a new perspective on the examination of molecules associated with obesity. Further investigation and confirmation of our results, involving a larger cohort of patients and additional parameters, are necessary to enhance our understanding of the impact of obesity on pregnancy.