Several aspects of flow have been shown to be important determinants of biological community structure and function in streams, yet direct application of this approach to large rivers has been limited. Using a multivariate approach, we grouped flow gauges into hydrologically similar units in the Missouri and lower Yellowstone Rivers and developed a model based on flow variability parameters that could be used to test hypotheses about the role of flow in determining aquatic community structure. This model could also be used for future comparisons as the hydrological regime changes. A suite of hydrological parameters for the recent, post-impoundment period (1 October 1966–30 September 1996) for each of 15 gauges along the Missouri and lower Yellowstone Rivers were initially used. Preliminary graphical exploration identified five variables for use in further multivariate analyses. Six hydrologically distinct units composed of gauges exhibiting similar flow characteristics were then identified using cluster analysis. Discriminant analyses identified the three most influential variables as flow per unit drainage area, coefficient of variation of mean annual flow, and flow constancy. One surprising result was the relative similarity of flow regimes between the two uppermost and three lowermost gauges, despite large differences in magnitude of flow and separation by roughly 3000 km. Our results synthesize, simplify and interpret the complex changes in flow occurring along the Missouri and lower Yellowstone Rivers, and provide an objective grouping for future tests of how these changes may affect biological communities.