For the near-infrared (NIR) spectral analysis of the concentration of blood glucose, the calibration accuracy can be affected because of the existing of outlier samples. In this research, a Monte-Carlo cross validation (MCCV) method is constructed for eliminating outlier samples. The human blood plasma experiment in vitro and the human body experiment in vivo were introduced to evaluate the MCCV method for its application effect in NIR spectral analysis of blood glucose. And the uninformative sample elimination method based on modified uninformative variable elimination (MUVE-USE) was employed in this study for the comparison with MCCV. The results indicated that, like the MUVE-USE method, the outlier samples elimination method based on MCCV could be used to eliminate the outlier samples which came from gross errors (such as bad sample) or system errors (such as baseline drift). In addition, the outlier samples from the random errors of uncertain causes which affect model accuracy can be eliminated simultaneously by MCCV. The elimination of multiple outlier samples is beneficial to the improvement of prediction accuracy of calibration model.