This study presents a method for effective pectin extraction from the laminae of three tobacco varieties as a means of biomass valorisation. Two pre-treatment methods (cold ethanol vs. accelerated solvent extraction [ASE] with ethanol) were compared for their capacities to produce a high pectin yield. Enzymatic extraction of pectin was also tested as a green extraction procedure and compared to the acid extraction approach. The optimisation experiments revealed that cold ethanol extraction followed by acid hydrolysis is the most convenient method for pectin extraction; the optimal set of conditions for hydrolysis were identified as 90°C, pH 1.5, and 4 h of extraction. Applying these optimised conditions to the three Nicotiana rustica tobacco varieties yielded pectin recoveries of 66.2%, 57.8%, and 56.7% from the NRT63, Bakoum Miena, and NRT61 samples, respectively. Tobacco pectins were found to have a medium molecular weight and low methoxy content. These results highlight the potential of tobacco residues as feedstock for to produce pectin with dietary applications.