The work presented deals with the application of Single Photon Ionisation - Time-of-Flight Mass Spectrometry (SPI-TOFMS) for the investigation of tobacco smoke. SPI-TOFMS is a modern analytical technique, which enables the simultaneous analysis of a large number of organic species in complex gas mixtures in real time. The paper is a summary of a PhD thesis (1) and seven research articles, which were recently published in various scientific journals (2-8). Consequently, more detailed information on particular aspects can be found in there. The experimental part covers two different approaches, and therefore, it is divided into two sub-sections. In the first one, the SPI-TOFMS is coupled to a pyrolysis furnace. The objective is to examine the thermal behaviour of tobacco under various controlled conditions. In so doing, three tobacco types (Virginia, Oriental, and Burley) were pyrolysed in two reaction gas compositions (nitrogen and synthetic air) and seven different furnace temperatures (400 °C, 500 °C, 600 °C, 700 °C, 800 °C, 900 °C, and 1000 °C). Results can help to unravel the complex formation and decomposition reactions taking place when tobacco is heated. In the second part the SPI-TOFMS is connected to a cigarette smoking machine in order to investigate the behaviour of cigarette smoke constituents on a puff-by-puff basis. The work incorporates the comparison of whole smoke and gas phase of cigarette smoke, a puff-resolved quantification of several hazardous smoke constituents, and the overall chemical characterisation of the individual smoking puffs. In addition, a critical consideration of the prevailing smoking procedure is given when applied to single puff analysis. A further study examines the influence of five different cigarette lighting devices (gas lighter, electric lighter, candle, match, and burning zone of another cigarette) on the chemical composition of the first puff.