In this study, we used Mid-Infrared Spectroscopy (MIRS) to analyze the biochemical fingerprints of nine termite colonies (soldiers, minor and major workers). We then examined whether these spectra could be used to differentiate termite colonies and if their differences could be explained by the geographical distances between colonies and/or their ages. We demonstrated that when only data from the heads of minor workers were considered, the specific fingerprint of each colony appeared to reflect its coordinate position in the study site. However, termite mound height, a proxy for colony age, was the main factor governing these signatures when data from the abdomen of major workers were used. Thus, this study shows the potential of MIRS for differentiating termite colonies in the field. It also highlights the close relationship between the physiological states of termite colonies and their environment.