Non-nutritive sweeteners (NNS) have been suggested as a viable solution to reduce sugar consumption. However, conflicting and uncertain evidence regarding their impact on lipid profile exists. This meta-analysis assesses the effects of different types of NNS on lipid profile markers. PubMed, Web of Science ISI, Scopus, and Google Scholar were searched for relevant articles up to September 2024. The Cochrane Collaboration tool was used for the quality assessment, and the evidence's trustworthiness was assessed using the Grading of Assessment, Development, and Evaluation of Recommendations (GRADE). A total of 28 articles revealed that NNS might not have a significant effect on serum triglycerides (Weighted mean difference (WMD): -2.66, 95% confidence interval (CI): -6.29, 0.98 mg/dL, n = 2209, level of confidence: low), total cholesterol (WMD: -0.44, 95% CI: -2.96, 2.08 mg/dl, n = 2047, level of confidence: low), high-density lipoprotein cholesterol (WMD: 0.03, 95% CI: -0.68, 0.74 mg/dl, n = 2097, level of confidence: low), low-density lipoprotein cholesterol (WMD: 0.51, 95% CI: -1.57, 2.59 mg/dl, n = 1777, level of confidence: low) and very low-density lipoprotein (WMD: 2.20, 95% CI: -2.66, 7.06 mg/dl, n = 154, level of confidence: very low). It is demonstrated that NNS might not affect serum lipid profile.