HPLC Determination of 3 Polyamines in Fresh Tobacco Leaves

The fresh tobacco leave sample was extracted ultrasonically by 0.4 mol·L^-1 HClO₄ solution, and then derived with using 4 g·L^-1 dansulfonyl chloride acetone solution as derivating reagent at 40℃ for 40 min. HPLC was applied to the determination of 3 polyamines, i.e. putrescine, spermidine and spermine in the sample solution. ZORBAX SB-C₁₈ chromatographic column (4.6 mm×150 mm, 5 μm) was used as separation column with mixtures of 0.01 mol·L^-1 ammonium acetate solution and acetonitrile with different mixing ratios as mobile phase in the gradient elution, and photodiode array detector was used for determination. Linear relationships between values of peak area and mass concentration of 3 polyamines were kept in the same range of 0.50-50.0 mg·L^-1, with detection limits (3S/N) in the range of 0.07-0.08 mg·L^-1 and lower limits of determination (10S/N) in the range of 0.23-0.28 mg·L^-1. The proposed method was applied to the analysis of samples of fresh tobacco leaves, giving values of recovery found by standard addition method and RSDs (n=6) of determined values in the ranges of 86.4%-100% and 0.70%-4.4% respectively.

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