Analysis of OPA-derivatized amino sugars in tobacco by high-performance liquid chromatography with fluorimetric...

An analytical method is developed to quantitatively determine glucosamine, galactosamine, and mannosamine in dried-and-ground burley and flue-cured tobaccos. Extraction is shown to be quantitative in the range of 0.01 to 2.0% (w/w). The extraction procedure consists of shaking one g of sample with 50 mL of deionized water adjusted to pH 7 for 30 min. This extract is filtered directly into an autosampler vial. An autosampler is programmed to withdraw two different aliquots: one with o-phthalaldehyde (OPA) derivatizing solution and the second one from the tobacco extract. The derivatization reaction occurs in the tubing connecting the autosampler and the chromatographic column. The OPA derivatives of these aminosugars are then detected with a fluorimetric detector, and their simultaneous analysis is performed with an external standard. This method is shown to be selective, accurate, and precise.