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The CHIRALPAK-IC was used, and the mixed solution of V (re-hexane) : V (isopropanol) = 95 : 5 was used as the mobile phase, and diethylamine was used as the mobile phase additive. In normal phase chromatography mode, a chiral column-high performance liquid chromatography-diode array detector (HPLC-DAD ) method was developed for the separation and analysis of nicotine optical isomers in tobacco and tobacco products. The characterization and test results of this method indicated:) The linear relationship between S-( -)-nicotine and R-( +)-nicotine was in the range of 15 ~750 µg/mL and 0. 75 ~30. 00 µg/mL, respectively. The detection limit and limit of quantification were 0.2 mg/g and 0. 05 mg/g, respectively. 2) The precision of intra- and inter-day measurements of nicotine was less than 0. 85% and 4. 00%, respectively; the recovery rates of S-( - ) -nicotine and R-( + ) -nicotine were 94. 93% ~ 103. 70% and 92. 50% ~ 117. 50%, respectively. This indicated that the method had good precision and high recovery rate. Using this method to determine the nicotine optical isomers of 26 different types of tobacco and tobacco products, it was found that tobacco was mainly S-( - )-nicotine, only 6 samples were detected by R-( + )-nicotine. The proportion of nicotine quality was 0. 18% ~0.70% ; among the 10 finished cigarettes and their mainstream smoke, the proportion of R-( + )-nicotine in total cigarette smoke was higher than that of cigarette tobacco. [ABSTRACT FROM AUTHOR]