In terms of the near-infrared spectroscopy, the concentration measurement for a substance in aqueous solution samples would be greatly influenced by the samples’ temperature since the water’s absorption in the solution will be obviously changed when the temperature varies. Especially for the measurement of glucose, which exhibits relatively weak absorption to the near-infrared light, the measurement accuracy would drop even with the slightest change of the solution’s temperature. It is found by experiment that the absorbance of glucose aqueous solutions would be changed in 0.344 7% at 1 160 nm when their temperature varies by 1 ℃ from around 30 ℃. Some recently developed methods based on chemometrics are presented, which can calibrate the solution’s spectra affected by temperature, to improve the glucose measurement accuracy under various temperatures. The methods include generalized least squares weighting (GLSW) and external parameter orthogonalisation (EPO). These methods are tested on the glucose aqueous solutions’ spectral data of 900~1 350 nm recorded at various temperatures, the prediction errors are evaluated using the spectra calibrated by the methods. The results show that the spectra of the same solution with various temperatures can be well calibrated to an equivalent result to the spectrum at the reference temperature. Moreover, the methods are compared with the multivariable regression of (partial least squares, PLS) regression, which includes temperature as a variable in the prediction model. The result shows that GLSW and EPO can well calibrate the solutions’ spectra from various temperature values because the coefficient of variation was reduced. And then the calibrated spectra show better performance on the prediction of glucose concentration than PLS because the complexity and the prediction errors of the models were well improved. The research in this paper could be referenced for the other substance measurements in the aqueous samples, and it can also be referredto the blood glucose measurement in human tissues.