Pancreatic surgery is a highly demanding and routinely applied procedure for the treatment of several pancreatic lesions. The outcome of patients with malignant entities crucially depends on the margin resection status of the tumor. In this study we describe the application of fiber-based attenuated total reflection infrared (ATR IR) spectroscopy for label-free discrimination of normal pancreatic, tumorous and pancreatitis tissue. The method was applied for the unprocessed freshly resected tissue samples of 40 patients, and a classification model for differentiating between the distinct tissue classes was established. The developed three-class classification model for tissue spectra allows the delineation of tumors from normal and pancreatitis tissues. The classification algorithm provides probability values for each sample to be assigned to normal, tumor or pancreatitis classes. The established probability values were transferred to a Red-Green-Blue (RGB) color plot. Subsequently, the method was translated into intraoperative application. Fiber optic ATR IR spectra were obtained from freshly resected pancreatic tissue directly in the operating room. The spectroscopic findings could subsequently be confirmed by the histology gold standard. This study shows the possibility of applying fiber-based ATR IR spectroscopy in combination with a supervised classification model for rapid pancreatic tissue identification with a high potential for transfer into intraoperative surgical diagnostics.