Simultaneous Determination of Six Organic Acids in Tobacco with Reverse-phase High Performance Liquid Chromatography

A method for simultaneous and rapid determination of 6 organic acids (including citric, malic and oxalic acid) in tobacco with reverse-phase high performance liquid chromatography (HPLC) was developed.HPLC was performed on a Hypersil C18 column (250mm×4.6mm, 5μm) at 25℃, using 2% CH₃OH-98%NaH₂PO₄(0.01mol/L;pH2.98) as mobile phase at the flow rate of 1.0mL/min and the detection wavelength of 210nm.The recoveries of the organic acids ranged from 98.2%to 104.3%with relative standard deviations of 0.61%-3.38%, and the limits of detection were 0.04-0.53μg/mL (S/N=3).The runtime was less than 7min.The 6 organic acids in cured tobacco leaves of K326 covered with four sun light-conversion films and one ordinary film were also analyzed.