Abstract: In order to clarify the differences of aroma components between the characteristic note tobacco mutant (Samson) and Zhongyan 100 (ZY100, the control) and to optimize the detection of volatile aroma components in tobacco, a headspace-solid phase microextraction arrow and gas chromatography-mass spectrometry (HS-SPME arrow-GC/MS) method was established by investigating the type of SPME arrow, equilibrium time, extraction time and extraction temperature. The aroma components in two tobacco materials were analyzed by this method, and the characteristic aroma components in Samson tobacco mutant were analyzed by principal component analysis (PCA) and partial least squares-discrimination analysis (PLS-DA). The results showed that:1) The optimized extraction conditions of SPME arrow were 120 μm×20 mm CAR/PDMS, equilibrium for 40 min and extraction for 40 min at 70℃. Under the conditions, the number of peaks with a relative standard deviation of less than 30% in the quality control (QC) samples accounted for 89.3% of the total peaks. 2) A total of 33 aroma components were detected in the two materials, among them the contents of 29 components in Samson tobacco mutant were higher than those in ZY100. This method has the advantages of easy operation, good reproducibility, stability and reliability, it is suitable for analyzing the characteristic aroma components in tobacco leaves.