This study was described for measuring clinically relevant levels of glucose in undiluted plasma and whole blood by near-infrared (NIR) spectroscopy. Result from an initial measurement of major blood components powder was over-lapped the absorption bands of glucose at 1500-1600 nm. However, the NIR data of blood components were clearly separated by principle component analysis (PCA) space. By the use of partial least squares (PLS) regression, glucose concentrations in undiluted plasma and whole blood could be determined with standard errors of prediction (SEP) of 15 mg/dl and 76 mg/dl, respectively. Although these blood components possessed strong absorption bands that overlapped with the absorption bands of glucose, successful calibration models could be carried out.