Bidi cigarettes, or bidis, are a tobacco product that originated in India and have been gaining popularity in the USA during the past few years, particularly with adolescents. As with conventional cigarettes, tobacco and smoke from bidis contain chemical constituents including carcinogenic chemicals such as the tobacco-specific nitrosamines (TSNAs). To help better assess the potential public health risk associated with bidi cigarettes, we developed modern high throughput methods to accurately quantify TSNA levels in tobacco and mainstream cigarette smoke particulate. We determined the TSNA levels in the tobacco filler and mainstream smoke from 14 bidi cigarette brands. In the bidi tobacco filler, the 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) levels ranged from 0.09 to 0.85 microg/g, while N'-nitrosonornicotine (NNN) levels ranged from 0.15 to 1.44 microg/g. These amounts are comparable with those in typical American blended cigarettes. The levels of NNK in mainstream smoke from bidis ranged from 2.13 to 25.9 ng/cigarette, and NNN levels ranged from 8.56 to 62.3 ng/cigarette. The wide variation in the TSNA levels most probably reflects the hand-rolled nature of the bidi cigarettes, resulting in a product with less homogenous tobacco amount and a wider variation in overall cigarette construction quality. TSNA levels of bidis were comparable with those of conventional cigarettes, and bidis should not be considered a lower-risk alternative tobacco product. Our analytical findings concur with the previous biologic and biochemical evidence supporting epidemiologic studies linking bidi use with various cancers, especially oral cavity and lung cancers.