Schizophrenia (SCH) is a serious psychiatric disorder characterized by positive symptoms, negative symptoms, and cognitive impairments, leading to long-term disability and invalidity. Therefore, the need for reliable biomarkers for its early diagnosis and treatment is crucial. This study aimed to conduct untargeted lipidomics of serum samples from a Serbian cohort, consisting of 30 schizophrenia patients and 31 non-psychiatric control individuals, using liquid chromatography (LC) combined with high-resolution mass spectrometry (HRMS) and chemometric analyses. Principal component analysis (PCA) of all samples showed no distinct separation between SCH and control groups but revealed clear gender separation within the control group. Multivariate statistical analyses (PCA and orthogonal partial least squares discriminant analysis OPLS-DA) of gender-differentiated SCH and control groups identified forty- nine differential lipids distinguishing male SCH (SCH-M) patients from male controls (C-M), and sixty potential biomarkers differentiating female SCH patients (SCH-F) from female controls (C- F). The lipidomic analysis of gender-differentiated groups (SCH-M vs. C-M and SCH-F vs. C-F) confirmed altered lipid metabolism, with decreased levels of the most affected lipid classes: glycerophospholipids (GP), sphingolipids (SP), glycerolipids (GL), and fatty acids (FA), compared to controls. Among the differential lipid metabolites with higher content in both SCH-M and SCH- F patient groups compared to their non-psychiatric controls, four common lipid molecules were identified: ceramides Cer 34:2 and Cer 34:1, lysophosphatidylcholine LPC 16:0, and triacylglycerol TG 48:2. The notable changes in lipid metabolism highlight the crucial role of metabolic pathways in the development of schizophrenia.