This study proposes a new preprocessing technique that combines Chebyshev filtering with baseline correction technique Asymmetric Least Squares (ALS) and Savitzky-Golay transformation (SGT) to improve the prediction of Glucose from near Infrared (NIR) spectra through linear regression models Partial Least Squares (PLS) and Principal Component Regression (PCR). To investigate the performance of the proposed technique, a calibration model was first developed and then validated through prediction of Glucose from NIR spectra of a mixture of glucose, urea, and triacetin in a phosphate buffer solution where the component concentrations are within their physiological range in blood. Results indicate that the proposed technique improves the performance of both PLS and PCR and achieves standard error of prediction (SEP) as low as 12.76 mg/dL which is in the clinically acceptable level and comparable to the existing literature.