Abstract Qinghai-Tibet Plateau is the highest plateau in the world, reputed as “the roof of the world” and “the world’s third pole”. Its ecological environment is characterized by low atmospheric oxygen pressure, cold, and limited feed supplies. This study aimed to investigate the serum immunoglobulin G (IgG) concentration and metabolic profiles of calves in high altitude area (3,650 m) fed different volumes of colostrum [8, 12, or 15% of birth body weight (BW)] using untargeted metabolomics strategies. Neonatal Holstein hybrid heifer calves (n = 44) were randomly divided into 1 of the 3 colostrum feeding amounts: 8% of birth BW (Low volume, L), 12% of birth BW (Middle volume, M), and 15% of birth BW (High volume, H) in colostrum. To safeguard the efficiency of colostrum intake, the colostrum was fed to calves within 0.5 h and 6 h after birth. The serum samples at 24 h after birth were collected for IgG concentration determination by indirect enzyme-linked immunosorbent assay (ELISA) and for LC-MS/MS metabonomic analysis. As the volumes of colostrum increased, there was a linearly increased in the serum IgG concentration (P < 0.01). Differential metabolites were selected according to the following 3 conditions: minimum fold change (FC) > 2, P-value < 0.01 in single-dimensional statistical analysis, and variable importance in projection (VIP) > 1 in the OPLS-DA model. Multivariate statistical analyses were performed to visualize differences among groups. Based on the screening criteria, a total of 3 differential metabolites (22 alpha-Hydroxy-campesterol, N-Myristoyl Histidine, and Calcitetrol) were detected between L and M, of which all differential metabolites were down-regulated. Six differential metabolites were detected between M and H, of which PC[P-18:1(11Z)/18:1(12Z)-2OH(9,10)] was up-regulated and 5 differential metabolites including 1-arachidonoyl-2-hydroxy-sn-glycero-3-phosphate, propanoyl phosphate, ascorbic acid 6-palmitate, 20-hydroxyecdysone, and 6’’’-deamino-6’’’-hydroxyparomomycin II were down-regulated. There were 28 differed metabolites (including leucyl-tryptophan, calcitetrol, 3’’-Oxoribostamycin) between L and H, of which 9 differential metabolites were up-regulated and 19 differential metabolites were down-regulated (Figure 1). Based on KEGG metabolic enrichment pathway analysis and pathway topology analysis, porphyrin metabolism was identified as the most impacted pathway between L and H (Figure 2). In conclusion, increasing colostrum volumes significantly increased serum IgG concentrations at 24 h after birth, favoring the success of the transfer of passive immunity. Alterations in serum metabolome of calves in the Qinghai-Tibet Plateau fed different volumes of colostrum were successfully revealed by untargeted metabolomics. The role of the identified differential metabolites and enriched pathway in the transfer of passive immunity in calves at high altitude deserves further investigation. Keywords: calf, colostrum volumes, high altitude area