Determination of sugars in tobacco leaf by HPLC with evaporative light scattering detection

A reversed phase liquid chromatography-evaporative light scattering detection, combined with solid phase extraction method was developed for the separation and determination of sugars in tobacco leaf. Separation was performed with a Hypersil NH2 analytical column and the mobile phase 75%:25% acetonitrile-water. In the range of 0.1-4 mg/mL, the method showed good linearity, with the correlation coefficients exceeding 0.99. The limits of detection of xylose, fructose, glucose, sucrose, maltose, and raffinose were 2, 2, 3, 2, 4, and 4 mu g/mL, respectively. The relative standard deviations of peak areas for these sugars were in the range of 2-3%. Recoveries of fructose, glucose, and sucrose were between 96 and 117%. Finally, determination of sugars in tobacco leaf, before and after bio-catalysis, was performed with this method.