Consumption of turmeric diets demonstrated preventive effects on high-fat diet (HFD) induced hyperlipidaemia. However, the preventive effects on serum lysophosphatidylcholines (Lyso PCs) have not yet been studied. For this study, an approach for characterization of Lyso PCs in mice serum using LC-QTOF-MS/MS was developed. The preventive effects of turmeric diets were evaluated on quantitative changes of individual Lyso PCs in mice serum by LC-QTRAP-MS/MS. As a result, 74 Lyso PCs were characterized, among which 59 Lyso PCs were quantified. The results indicated that significant changes occurred for the Lyso PCs with higher degree of unsaturation in the fatty acid chain. This is the first report regarding a targeted metabolomic approach for analyses of serum Lyso PCs and its application to interpretation of preventive effects of turmeric on mice hyperlipidaemia. These metabolic changes and potential biomarkers might serve as scientific evidence for future diagnosis and therapeutic intervention of hyperlipidaemia.