Although airborne pollutants in urban buses have been studied in many cities globally, long-distance buses running mainly on highways have not been addressed in this regard. This study investigates the levels of volatile organic compounds (VOCs), carbon monoxide (CO), carbon dioxide (CO2) and particulate matter (PM) in the long-distance buses in Taiwan. Analytical results indicate that pollutants levels in long-distance buses are generally lower than those in urban buses. This finding is attributable to the driving speed and patterns of long-distance buses, as well as the meteorological and geographical features of the highway surroundings. The levels of benzene, toluene, ethylbenzene and xylene (BTEX) found in bus cabins exceed the proposed indoor VOC guidelines for aromatic compounds, and are likely attributable to the interior trim in the cabins. The overall average CO level is 2.3 ppm, with higher average level on local streets (2.9 ppm) than on highways (2.2 ppm). The average CO2 level is 1493 ppm, which is higher than the guideline for non-industrial occupied settings. The average PM level in this study is lower than those in urban buses and IAQ guidelines set by Taiwan EPA. However, the average PM10 and PM2.5 is higher than the level set by WHO. Besides the probable causes mentioned above, fewer passenger movements and less particle re-suspension from bus floor might also cause the lower PM levels. Measurements of particle size distribution reveal that more than 75% of particles are in submicron and smaller sizes. These particles may come from the infiltration from the outdoor air. This study concludes that air exchange rates in long-distance buses should be increased in order to reduce CO2 levels. Future research on long-distance buses should focus on the emission of VOCs from brand new buses, and the sources of submicron particles in bus cabins.