Filter ventilation in cigarettes may alter smoking behavior and impact exposure to harmful chemicals. This study examined the effect of filter ventilation on smoking topography and urinary levels of metabolites of known tobacco-associated toxicants. Twelve male daily Marlboro Red (MR) cigarette smokers (aged 26-34) participated in the study. In a controlled environment, participants smoked regular MR, Marlboro Prime (MP, a low tar, nicotine, carbon monoxide version of MR), or ventilation holes-blocked MP (MPT; where T = taped) ad libitum on separate days. Smoking topography was recorded by using the CReSSmicro device. Urine samples were collected throughout the day, and 27 metabolites of tobacco-associated toxicants (i.e., nicotine, aldehydes, xylene) were analyzed by ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS). Smoking different types of cigarettes throughout the day resulted in increased urinary metabolites, including those of nicotine, aldehydes, xylene, and others. No differences were observed in intraday increases in these metabolites between smoking MR and MP. However, compared with MR and MP, smoking MPT was associated with a smaller increase in urinary concentrations of cotinine and 3-hydroxycotinine. Switching from the MR to MP or to the MPT did not significantly alter the number of cigarettes smoked. However, puff count, puff duration, and total cigarette volume smoked were significantly lower when smoking MPT. Overall, our data supports existing literature that indicates that smoking high filter-ventilated cigarettes elicits equivalent toxin exposure as conventional cigarettes. Blocking ventilation holes led to lower urinary nicotine metabolite concentrations, which may be partly due to the rapidly altered smoking topography.