Aim To analyze the changes of lipid metabolites in menopausal rats and identify potential biomarkers in metabolic changes. Methods 4-month-old female SD rats were randomly divided into sham operation (Sham) group and ovariectomized (OVX) group. Using UPLC/LTQ-Orbitrap-MS metabonomics analysis method, principal component analysis (PCA) and orthogonal-partial least squares analysis (OPLS-DA) were used for multivariate statistical analysis to screen potential biomarkers related to diseases. Results The OPLS-DA score plot could be used to distinguish the plasma lipid metabolites of rats in the sham group and ovariectomized group, and there was no significant overlap between the two groups. Compared with the sham group, the relative contents of ceramide (Cer), sphingomyelin (SM), platelet activating factor (PAF), lysophosphatidylcholine (LPC), triglyceride (TG) and omega-6 unsaturated fatty acid (ω-6PUFA) increased in the ovariectomized group, while the relative contents of diglyceride (DG), phosphatidylglycerin (PG) and phosphatidylserine (PS) decreased. A total of 35 endogenous metabolites were detected in both groups, including Cer, SM, PAF, LPC, TG, DG, PS, PG, ω-6PUFA as potential biomarkers. Conclusion All the 9 potential biomarkers were related to menopausal syndrome, which laid a foundation for the early detection of biomarkers of senile degenerative diseases.