This study was carried out to improve the analytical methods for determination of organic acids in tobacco leaf by HPLC. The samples for HPLC analysis were pre-treated by means of SPE. The calibration curve for each acid was linear and $R^2$ values ranged from 0.9990 to 1.0000. The limit of detection were obtained from the calibration curves and their values were between 1.39 to $4.87{mu}g/mL$. Recovery rates of organic acids were between 88.6 % to 98.5 %. The concentrations of organic acids among the various tobacco leaves were compared to the concentration of organic acids, were in the order oriental, burley, flue-cured tobacco. In the case of flue-cured and oriental tobacco leaves, the order of concentration of organic acids was malic acid, oxalic acid, citric acid. But in the case of burley tobacco leaves, the order of concentration of organic acids was citric acid, oxalic acid, malic acid.