In this paper, employed an inter-standard quantitative method for determination of benzo[a]pyrene and its structural isomers (B[b]F, B[k]F, B[j]F, B[j]A, B[e]A, B[e]P, PER) in mainstream cigarette smoke by GC-MS/MS. The results showed that the LODs and the LOQs of target PAHs could achieve 0.056 6~0.181 7 and 0.188 6~0.605 5 ng/cig, respectively, while the linear correlation efficiencies are larger than 0.998 for all PAHs. The Intra-assay and Inter-assay precisions measured by 15 repeat runs are 1.91%~3.97% and 2.54%~4.46%, respectively. And with three different added-levels, the recovery rates for the target PAHs lie between 92.60% and 107.60%. It was proved that the developed method is environmentally friendly, highly sensitive and selective.