The metabolic effect of Fo-Shou-San on blood deficiency mice was studied by using metabolomic method.  UPLC-QTOF/MS was used to analyze the plasma metabolome in blood deficiency mice.  MS data were processed by MarkerLynx software.  With multivariate statistical analysis of plasma metabolite profiles, a clear separation among control, blood deficiency model, and Fo-Shou-San groups was achieved.  Potential biomarkers were selected according to the parameters of variable importance in the projection (VIP) and identified according to MS information and database retrieval.  The metabolic network of blood deficiency was predicted via MetPA database.  Twenty-two potential biomarkers were identified and used to explain the thiamine metabolism, arachidonic acid metabolism, sphingolipid metabolism, glyoxylate and dicarboxylate metabolism, histidine metabolism, nicotinate and nicotinamide metabolism, cysteine and methionine metabolism, tryptophan metabolism, starch and sucrose metabolism, tyrosine metabolism and citrate cycle (TCA cycle).  Those metabolic pathways were disturbed in blood deficiency mice, but which could be regulated nearly to normal state after Fo-Shou-San administration.  In this study, the metabolomics of blood deficiency mice and the action mechanism of nourishing blood effect of Fo-Shou-San were evaluated.  The physiological and metabolic state of the organism could be represented comprehensively by using metabolomics.  And metabolomics can be used to evaluate the pharmacodynamics and related mechanisms of Chinese medicine and formulae.