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The analytes were resolved on an Acquity UPLC high strength silica (HSS) perfluorophenyl (PFP) (150 mm 2.1 mm, 1.8 m) column (Waters Inc., MA) maintained at 40C, using a binary gradient consisting of 0.2% formic acid (Solvent A) and methanol (Solvent B)

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The specific types and concentrations of pyrazines produced depend on factors such as: Temperature and humidity during curing Duration of the curing process Tobacco variety and growing conditions Processing methods used by manufacturers Storage conditions before cigarette production Many major tobacco brands incorporated pyrazines to enhance product appeal and increase sales

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