By directly measuring human soluble blood samples by Fourier transform infrared spectroscopy (FTIR) and attenuated total reflection (ATR) technology, the rapid quantitative analysis method for human hemoglobin (HGB) was established. The optimization of Savitzky-Golay (SG) smoothing modes combined with partial least squares (PLS) factor was applied to optimize the model of FTIR/ATR spectroscopy analysis here. The optimal smoothing parameters were 1st order derivative smoothing, 2nd degree polynomial, 59 smoothing points, the optimal PLS factor, RMSEP, RP and RRMSEP were 8, 4.21g/L, 0.962 and 3.1% respectively, which was obviously superior to the direct PLS model without SG smoothing. This demonstrated that FTIR/ATR spectroscopy can be applied to the rapid determination of human hemoglobin concentration, large-scale simultaneous optimization selection of SG smoothing parameters and PLS factor can be effectively applied to the model optimization of FTIR/ATR analysis.