Provided is a method for detecting 
nicotine optical isomers in buccal tobacco by using normal-phase liquid 
chromatography-
tandem mass spectrometry. The method is characterized in that firstly, 
nicotine in the buccal tobacco is extracted, a chiral 
chromatographic column is adopted to separate the 
nicotine optical isomers, a post-column high-pressure 
infusion pump introduces isopropanol and conducts splitting, 
assay determination is conducted in combination with the 
tandem mass spectrometry, and therefore the sensitive detection and the separation of S-(-)-nicotine and R-(+)-nicotine are achieved. The method has the advantages that by the adoption of the post-column high-pressure 
infusion pump for introducing the isopropanol and conducting the splitting, the volume of a non-polar 
solvent added into a 
mass spectrometer is reduced, the volume of the polar 
solvent isopropanol is compensated at the same time, the 
ionization efficiency is improved, so that requirements of liquid 
chromatography-
tandem mass spectrometry analysis are met, and the sensitivity of the analysis is improved; compared with current methods, the 
chromatographic separation degree of the two nicotine optical isomers is high, wherein 
retention time difference between the two nicotine optical isomers is one minute; the analysis time is short, wherein only seventeen minutes are needed.