A study design procedure was developed and demonstrated for the deployment of portable onboard tailpipe emissions measurement systems for selected highway vehicles fueled by gasoline and E85 (a blend of 85% ethanol and 15% gasoline). Data collection, screening, processing, and analysis protocols were developed to assure data quality and to provide insights regarding quantification of real-world intravehicle variability in hot-stabilized emissions. Onboard systems provide representative real-world emissions measurements; however, onboard field studies are challenged by the observable but uncontrollable nature of traffic flow and ambient conditions. By characterizing intravehicle variability based on repeated data collection runs with the same driver/vehicle/route combinations, this study establishes the ability to develop stable modal emissions rates for idle, acceleration, cruise, and deceleration even in the face of uncontrollable external factors. For example, a consistent finding is that average emissions during acceleration are typically 5 times greater than during idle for hydrocarbons and carbon dioxide and 10 times greater for nitric oxide and carbon monoxide. A statistical method for comparing on-road emissions of different drivers is presented. Onboard data demonstrate the importance of accounting for the episodic nature of real-world emissions to help develop appropriate traffic and air quality management strategies.