Abstract: A method for determining seven alkaloids, including free-base nicotine, nornicotine, myosmine, nicotyrine, anatabine, 2, 3'-pyridine and cotinine in the saliva of cigarette smokers was developed by coupling headspace(HS)solid-phase microextraction(SPME) with gas chromatography tandem mass spectrometry(GC-MS/MS). Human saliva samples were collected with Salivette^@ collection kit and the quantification was achieved by using quinoline as the internal standard. Six samples of human saliva were analyzed. The results showed that:1) The experimental conditions were 30 μL saliva samples and white extraction head, using elastic quartz capillary column in the measurement of the selected ion monitoring mode for GC/MS analysis, extracting for 40 minutes at 80℃, and desorbing for 3 minutes. 2)The relative standard deviations(RSD) for the seven alkaloids were between 1.71% and 4.52%, the recovery ranged from 95.1% to 103.2% with the linear correlations above 0.991. 3)The main alkaloids in the saliva were free-base nicotine and nornicotine, the content of nicotine was 0.445 8-0.742 2 μg/mL, and that of nornicotine was 1.371 3-2.854 9 μg/mL. This method is simple, stable, accurate, and suitable for the simultaneous determination of seven alkaloids in the saliva of smokers.