The Ultra Performance Liquid Chromatography-Triple-Time of Flight Mass Spectrometer (UPLC-Triple-TOF-MS/MS) based platform was developed to investigate the serum metabolic profile associated with the growth performance in Chinese softshell turtle fed with the diet added with Traditional Chinese Medicines (TCMs) feeding additive. 0.0%, 1.0%, 2.0%, 3.0%, 4.0% of TCMs were added to the base ration of the control group (CG), TCMs treatment group1 (TG1), TCMs treatment group2 (TG2), TCMs treatment group3 (TG3) and TCMs treatment group4 (TG4), respectively. Compared with control group (CG), the weight gain and special growth ratio firstly increased and lately decreased with increased dietary TCMs levels. The highest weight gain (5.824 ± 1.205) and specific growth rate (3.887 ± 0.367) were found in the TCMs treatment group 2 (TG2) (P < 0.05), and the lowest feed conversion ratio (1.785±0.015) also appeared in the TG2 group (P < 0.05). In the metabolomics assay, a total of 795 (VIP> 1) out of 3089 variable metabolites were significantly different between the TG2 and CG. 43 potential biomarkers between TG2 and CG groups were finally screened out on the basis of VIP > 1 and P < 0.05. The potential networks and pathways for the model were explored using MetaboAnalyst. A total of 43 potential biomarkers were confirmed involving thirteen different metabolic pathways. Following the diet added with 2% TCMs feeding additive, five metabolic pathways were affected, these included the following: phenylalanine, tyrosine and tryptophan biosynthesis, tyrosine metabolism, cysteine and methionine metabolism and arginine biosynthesis. This study indicated that the dietary supplementation TCMs as feed additives could influence the growth performance of the healthy juvenile turtles, and the optimal dose of supplementing TCMs to juvenile turtleswas 20 g/kg in diet.