The aim of this study was therefore to determine whether advanced multivariate analysis of bile spectral profiles obtained by FTIR spectroscopy may differentiate bile samples from patients with and without bile duct malignant strictures. In order to reduce the overlapping spectral signals due to complex molecular composition of bile, we performed, in addition to the whole bile analysis, a separate analysis of the aqueous and the organic phases of bile using a lipid extraction procedure. Sensitivity and specificity for the three bile samples were computed and the overall results compared and discussed.