Five Pu-erh tea samples with stale flavor were selected from 36 samples by sensory tests, and the aroma constituents of these samples were determined by HS-SPME/GC-MS. Furthermore the mainly difference in aroma constituents between Pu-erh tea with stale flavor and other Pu-erh tea samples was discussed in the paper. Results showed that the aroma constituents of Pu-erh tea with stale flavor were mainly heterocyclic oxygen compounds and alcohols, and the sum of the two mentioned above occupied 40% of the total, the major aromatic compounds of Pu-erh tea with stale flavor were 1,2,3-Trimethoxybenzene, Benzene,4-ethyl-1,2-dimethoxy-,β-Terpineol, Epoxylinalol, Linalool oxideⅠ, 3,4-Dimethoxytoluene, Benzene, 1,2-dimethoxy-, Cedrol, Butylated Hydroxytoluene, Benzene, 1,2,3-trimethoxy-5-methyl-, β-Linalool, 1,2,4-Trimethoxybenzene, β-Ionone, α-Farnesene. The contents of β-Linalool, Decanal, Nonanal, Methyl Salicylate, 3,4-Dimethoxytoluene, Benzene, 4-ethyl-1,2-dimethoxy-, Butylated Hydroxytoluene were much higher than that of other Pu-erh tea samples. However, the contents of β-Terpineol, Cedrol, β-ionone, 1,2,4-Trimethoxybenzene, Benzene, 1,2-dimethoxy-, 1,2,3-Trimethoxybenzene and Longifolene were much lower than that of other Pu-erh tea samples.