Determination of Sugars in Tobacco Leaf by HPLC with Evaporative Light Scattering Detection

Abstract 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 µg/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.