As one of the most common endocrine diseases, diabetic mellitus(DM) has a very close relationship with phospholipids metabolism. Tangshen Formula(TSF) is an often-used prescription with good efficacy for treating DM and diabetic nephropathy(DN). The aim of this study is to understand the effect of TSF on phospholipids metabolism of DM. In this paper, Otsuka Long-Evans Tokushima Fatty( OLETF) rats, a model of slowly progressive obese type II diabetes, were employed. Phospholipids profiling of rat plasma was established using normal phase liquid chromatography-time of flight mass spectrometry (NPLC-TOF/MS). Partial least-squares discriminant analysis(PLS-DA) and quantification of 14 compounds were applied to evaluate the effect of TSF on phospholipids metabolism of OLETF rats and to discover potential biomarkers. The results showed that plasma phospholipids metabolic disorders occurred in OLETF rats. The good effect of TSF on phospholipids metabolism was compared with the adverse impact of Monopril. TSF could significantly improve phospholipids metabolism of OLETF rats, whereas Monopril exacerbated disturbance of phospholipids metabolism. According to the quantitative results, a total of 8 phospholipids compounds which have significant concentration variation between control group and model group were considered as potential biomarkers of type II diabetes.