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Regional transportation systems face multiple challenges, including inadequate service quality and scarce and geographically dispersed demand. The Physical Internet (PI) concept offers potential solutions to these issues, yet its focus has primarily been on urban and freight-oriented contexts, overlooking the unique challenges of regional transportation. Moreover, the limited analysis of regional transportation has resulted in a lack of detailed understanding of demand dynamics and difficulties in aligning supply capabilities with demand expectations. To address these issues, this paper proposes a methodology for analyzing regional transport demand, that embraces the PI concept by treating both freight and passenger demand within a unified framework. The methodology defines user profiles, characterizes demand in terms of volume, time, and space, and identifies the factors influencing transportation use. An experimental study using the ECOTRAIN project validates the effectiveness of this approach. This case study not only shows the applicability of the methodology but also highlights its potential to guide the development of tailored transportation solutions suited to regional areas.