Abstract: In order to illustrate the differences in volatile components of various types of tobacco, a newly developed two-step headspace sampling-gas chromatography-mass spectrometry method was applied to analyze flue-cured tobacco, cigar tobacco, burley, oriental and sun-cured tobacco. The two-step headspace sampling system involved a coupling of the headspace sampler and a programmed temperature vaporizer, which adopted a purposely made large-volume loop to increase the method sensitivity, and then a programmed temperature vaporizer to avoid the peak broadening caused by the large injection volume. The results showed that: 1)a total of 121 volatile components were detected from the five types of tobacco; 2)the principal component analysis and hierarchical cluster analysis were performed to verify the relationship between the volatile compositions and the tobacco type; 3)the volatile components of sun-cured and oriental tobacco was the most similar, while the cigar tobacco was different from all the other types of tobacco; 4)the composition of volatile components of the cigar tobacco was closely related to its leaf origins. In conclusion, the new method is simple, sensitive and reliable, and could be used to evaluate tobacco components and tobacco quality.