Identification and quantitation of glycosidically bound aroma compounds in three tobacco types by gas chromatography–mass spectrometry

• New method for qualitative and quantitative analysis of glycoside in tobacco samples.
• 21 Bound aglycones were identified by β-glucosidase hydrolysis.
• 15 Glucosides and 1 rutinoside were tentatively identified and quantified by GC–MS.
• The PCA showed different type of tobacco could be clearly differentiated by glycoside.

Glycosidically bound aroma compounds in three different types of tobacco were investigated. After isolation of extracts obtained by Amberlite XAD-2 adsorption and ethyl acetate elution, glycosides were analyzed after enzymatic hydrolysis by gas chromatography–mass spectrometry (GC–MS) or directly after trifluoroacetylated (TFA) derivatization by GC–MS in electron ionization (EI) and negative chemical ionization (NCI) mode. In total 21 bound aglycones were identified by β-glucosidase hydrolysis. These aglycones mainly consisted of C13-norisoprenoids, aromatic components and sesquiterpenoids. Additionally, with the aid of enzymatic hydrolysis, 15 β-d-glucopyranosides and 1 β-d-rutinoside were tentatively identified by TFA derivatization. TFA method was validated by repeatability and successfully employed to analyze different types of tobacco. Principal component analysis (PCA) was carried out on identified glycoside variables to visualize the difference between the tobacco types and the relationship between the glycoside variables and the tobacco types was established.