Proposed FDA regulations may require significant reduction of the nicotine content of tobacco products. Lowering the nicotine content may impact tobacco leaf surface chemistry that affects flavour. The objective was to compare the leaf surface chemistry profiles of high- and low-alkaloid Burley varieties/lines grown under standard and low-nicotine field practices. Six varieties/lines, one check and five low alkaloid lines, were sampled shortly after topping. Leaf surface exudate was collected via solvent extraction and analyzed by GCMS. Qualitatively, chemical profiles of trichome exudate of four Burley lines (LA TN 90LC, ITB 1501, J14 and J29) and the check (TN 90LC) were highly similar, consisting of α- and β-cembratriene-diols (CBT-diol isomers), apparent diterpenoid oxidation products, sugar esters (SEs) (minor) and free sugars. The low-alkaloid French line ITB 259 produced labdane-type diterpenes cis-abienol and labdene-diol, but not cembranoid diterpenes. Such diterpene composition is unusual for a Burley tobacco. TN90 LC and LA TN 90LC had similar surface chemistry profiles and abundance of total compounds under standard field practices. Sugar esters and oxidized products were reduced in the six varieties/lines under the low nicotine field practices. The mutations leading to low-alkaloid variant of TN 90LC did not impact the leaf surface chemistry. Low-alkaloid field practices likely lowered the quality of Burley tobaccos.