Spectral performance would be affected by many factors such as temperature, equipment parameters and so on. Humidity fluctuations may occur in practice because of varying weather conditions. The objective of this research was to find out whether the change of humidity would influence the near infrared spectrum of samples. In this trial, an airproof, humidity-controllable test-bed was established to change the humidity of the mini environment. At 40%, 50%, 60%, 70% and 80% degrees of humidity, each sample's final spectrum was attained by removing the background's spectrum from the sample's. For whether the influence of the sample's and the background's spectrum are equal was not known, This trial was divided into two groups: detecting background and sample at each degree of humidity (group 1) and background's detecting just happened at 40% degree of humidity (group 2). This research was based on the hardware of NEXUS intelligent FT-IR spectrometer, made by Nicolet instrument company U.S.A, with using fiber optic diffuse reflectance accessory. The final spectrum was analysed using single variance analysis and Mahalanobis Distance methods. The result shows that neither in group 1 nor 2, humidity had little influence on NIR.