To discuss the feasibility of using the serum’s multi-optical path length spectroscopy information for measuring the concentration of the human blood components, the automatic micro-displacement measuring device was designed, which can obtain the near-infrared multi-optical path length from 0 to 4.0 mm (interval is 0.2 mm) spectra of 200 serum samples with multi-optical path length spectrum of serum participated in building the quantitative analysis model of four components of the human blood: glucose (GLU), total cholesterol (TC), total protein (TP) and albumin (ALB ), by mean of the significant non-linear spectral characteristic of blood. Partial least square (PLS) was used to set up the calibration models of the multi-optical path length near-infrared absorption spectrum of 160 experimental samples against the biochemical analysis results of them. The blood components of another 40 samples were predicted according to the model. The prediction effect of four blood components was favorable, and the correlation coefficient (r) of predictive value and biochemical analysis value were 0.932 0, 0.971 2, 0.946 2 and 0.948 3, respectively. All of the results proved the feasibility of the multi-optical path length spectroscopy technique for blood components analysis. And this technique established the foundation of detecting the components of blood and other liquid conveniently and rapidly.