Metabolism of lipids is essential for the regulation of a variety of key cellular functions. Recent advances in high performance liquid chromatography coupled with high resolution mass spectrometry have expanded our knowledge of lipid metabolism in diseases. Currently, sepsis is one of the most important public health problems all over the world, which is a serious systemic inflammatory syndrome leading to infection by various agents or trauma and subsequently to a multiple organ dysfunction response. However, little is known about the lipids affected by sepsis and their roles in kidney injuries. In this study, we present targeted and non-targeted lipidomics strategies to discover the lipid metabolism variation in serum in rats with sepsis-induced kidney injuries. Liquid chromatography (LC) coupled with mass spectrometry (MS) and multivariate data analysis were used to obtain the global lipid metabolic profiles. In addition, biochemical parameters and histopathological examination results for the kidney were analyzed to support the pathological changes during sepsis-induced kidney injury. The identification of ten proposed lipids and five relevant pathways will promote a better understanding of lipid profile alterations in kidney injury. The results suggested that lipid metabolism in sepsis-induced kidney injury had changed significantly and contribute by offering potential targets for clinical diagnosis and therapy in the future, which would be worth further studies to broaden the applications of high performance liquid chromatography coupled with high resolution mass spectrometry in the study of lipid metabolism.