Active transport contributes to increased daily physical activity (PA). Car ownership is associated with less frequent active transport and less PA. For business car ownership this relation is unknown. Therefore, we explored whether business car owners and their adult household members comply less with the Dutch moderate to vigorous physical activity (MVPA) guideline and are more sedentary than private car owners and persons without a car. From October 2011 to September 2012 questions about use and availability of cars in the household were included in the survey Injuries and Physical Activity in the Netherlands. Multiple linear regression was used to compare six mutual exclusive groups of ownership and availability of (business and/or private) cars in the household. Business car owners complied less (15.8 percent points) with the MVPA guideline than the other respondents. They also reported 1.5 h more sitting time during workdays than the other respondents, but after adjusting for covariates, this difference was no longer significant. We concluded that owners of a business car in the Netherlands are at higher risk of not complying with the MVPA guideline and tend to spend more hours sitting during workdays than other adults. Further research in this group, e.g. with objective instruments to measure physical activity and sedentary behavior, is recommended. Policy makers on transport and fiscal arrangements, employers, employees, occupational health professionals and car lease companies should be aware of this possible health risk.