Acceptor segregation is investigated as a function of compositional difference, Δx, between adjacent layers in (AlxGa1−x)0.5In0.5P heterostructures. Magnesium, Zn, Be, and Mn acceptor species are all shown to segregate out of the high band gap Al-rich (AlxGa1−x)0.5In0.5P layers and into the low band gap Al-poor (AlxGa1−x)0.5In0.5P layers during high temperature epitaxial growth of such heterostructures. The observed acceptor segregation appears to be independent of growth method or dopant incorporation method (metalorganic chemical vapor deposition, gas source molecular-beam epitaxy, or ion implantation), and increases with increasing compositional difference between adjacent (AlxGa1−x)0.5In0.5P layers. A theoretical model is developed to describe acceptor segregation based on charge separation and the resulting electric field across the heterointerface, and the resulting acceptor segregation is shown to vary as (mh AL*/mh CL*)3/2 exp(ΔEV/kT) where mh AL* and mh CL* are the hole effective masses in the active layer and confining layer, and ΔEV is the valence band offset. Comparison between experimentally measured and theoretically predicted acceptor segregation ratios gives excellent agreement for (AlxGa1−x)0.5In0.5P heterostructures over the range of compositional differences from Δx=0.12 to Δx=0.93.