Simultaneous determination of concentrations for four major components in human blood serum was investigated using a Fourier-transform mid-infrared spectroscopy. Infrared spectra of human blood serum were measured in 8.404 ∼ 10.25 ${mu}m$ range where the highest absorption peaks of glucose are located. A partial least square (PLS) algorithm was utilized to establish a calibration model for determining total protein, albumin, globulin and glucose levels which are commonly measured metabolites. The standard error of cross validation obtained from our multivariate calibration model was 0.24 g/dL for total protein, 0.15 g/dL for albumin, 0.17 g/dL for globulin, and 6.68 mg/dL for glucose, which are comparable with or meet the criteria for clinical use. The results indicate that the infrared absorption spectroscopy can be used to predict the concentrations of clinically important metabolites without going through a chemical process with a reagent.