Abstract
A simple and selective high performance liquid chromatographic (HPLC) method for the analysis of nicotine in a Hanks' balanced salts extract of smokeless tobacco components is presented here. An HPLC method was modified and applied for the analysis of nicotine in samples obtained from a biorelevant in vitro release system for permeation testing of nicotine from a commercially available smokeless tobacco product. The selectivity for the analysis of nicotine in vitro samples was investigated as a part of the method validation by the standard addition method, peak purity testing, and peak trapping followed by mass spectrometry (MS) identification of a co-eluant which was observed. The co-eluting compound was attributable to a leachable substance from the neoprene O-ring used in the apparatus. The mass spectrum (molecular ion with m/z 353.8 ≈ 354) for this substance was consistent with the molecular weight of the plasticizer, alkyl alkylether diester adipate (molecular weight 353), commonly used in manufacturing rubber. Selectivity was shown without chromatographic resolution of the co-eluting substance by selection of an appropriate wavelength range where the co-eluant did not absorb UV light.
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