The analysis of tobacco-related alkaloids such as nicotine and its metabolites presents many challenges for the analyst. The highly polar nature of these compounds limits the retention on common reverse phases such as the C18 phase. One major challenge caused by this lack of retention is the chromatographic resolution of several of the isobaric compounds is insufficient for LC-MS analysis. One solution to this is through the use of a different column phase such as the Thermo Scientific™ Accucore™ HILIC analytical column.
Hydrophilic interaction chromatography (HILIC) is an increasingly popular technique for separating polar compounds, such as tobacco alkaloids, which do not retain well using reversed-phase chromatography. For HILIC, a more polar mobile phase is the stronger elution solvent with water being the strongest elution solvent. This allows better retention of polar analytes than possible with reversed-phase separation conditions.
This technical note describes the use of a Thermo Scientific™ Accucore™ HPLC column using Core Enhanced Technology to facilitate fast and high efficiency separations. The 2.6 μm diameter particle technology is not fully porous, but instead features a solid core and a porous outer layer. The tightly controlled 2.6 μm diameter of the Accucore particles provides much lower backpressure than seen with sub-2 μm materials and are therefore an alternative format to traditional UHPLC. Analyte properties that govern retention with Accucore HILIC are acidity/basicity, which determines hydrogen bonding, and polarizability, which determines dipole-dipole interactions.
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