Objective: To evaluate the association between consumed non-nutritive sweeteners (NNS) and gestational diabetes mellitus (GDM) in a cohort of pregnant women from Santiago, Chile. Methods: This secondary data analysis of a cohort.involved 1,472 pregnant women from the Chilean Maternal-Infant Cohort Study-II (CHiMINCs-II). These women received care at primary health care centers in Puente Alto county, South-Eastern Metropolitan Health Service of Santiago, Chile. NNS consumption was estimated using 24-h dietary recalls and linked to the packaged foods nutrition facts panel. Plasma glucose values were extracted from clinical records. GDM was defined according to national criteria: 1) fasting plasma glucose (FPG) >100 and <126 mg/dL at the first antenatal visit; 2) FPG >100 mg/dL or 2-hour plasma glucose >140 mg/dL in the 75 g oral glucose tolerance test at 24-28 weeks. Cases with a GDM diagnosis in their medical records were also considered regardless of test results. The association between each NNS and GDM was assessed using logistic regression models. Results: A total of 77.8% of the participants consumed NNS. The most consumed was sucralose (66%), followed by acesulfame-K (43.6%), and steviol glycosides (41.1%). Beverages (82%), dairy (12.4%) and candy products (4.4%) were the primary dietary sources of NNS. The GDM incidence was 18.9%, higher among consumers of any NNS compared to non-consumers (20.3% vs. 14.2%, p < 0.05). The adjusted model showed a significant association between the consumption of any NNS and sucralose and the risk of GDM (OR for any NNS = 1.58; 95% CI: 1.10-2.26; P = 0.014; OR sucralose = 1.44; 95% CI 1.06-1.95; P = 0.020). Conclusions: The consumption of NNS, particularly sucralose, is associated with an increased risk of GDM in pregnant women. Further studies are essential to validate these results in other contexts and to guide future recommendations for healthier dietary practices among pregnant populations. (c) 2024 Elsevier Inc. All rights are reserved, including those for text and data mining, AI training, and similar technologies.