Porcine pancreatin extract (PPE), comprising porcine pancreatic lipase (PPL), α-amylase (PPA), and proteases, mediates the digestion of fats, carbohydrates, and proteins and is widely used to treat pancreatic insufficiency. N-glycosylation is a critical post-translational modification that contributes to enzyme stability and activity. This study profiles N-glycans in PPE, PPL, and PPA to assess their roles in enzymatic function. N-glycans were enzymatically released, labeled with procainamide, and analyzed by liquid chromatography–tandem mass spectrometry. Relative quantity was expressed as a percentage of the total N-glycan peak area, and absolute quantity was calculated using a calibration curve. We identified 32, 31, and 10 N-glycan structures in PPE, PPL, and PPA, respectively, including sialylated and fucosylated glycans. PPE (25.6%) and PPL (29.3%) showed higher sialylation than PPA (10.9%), whereas fucosylation accounted for 67.4%, 66.9%, and 54.3%. Total N-glycan quantities were 422, 602, and 131 pmol/mg in PPE, PPL, and PPA, respectively. Functional assays following deglycosylation, desialylation, and defucosylation showed that sialylation and fucosylation influence PPE and PPL activities, with minimal effects on PPA. These results show that sialylation and fucosylation are essential for PPE and PPL, but not PPA. This study provides the first N-glycan profiles of PPE, PPL, and PPA, highlighting their roles in structural stability and enzymatic regulation.