Layer-release techniques for producing freestanding III-V epitaxial layers have been actively developed for heterointegration of single-crystalline compound semiconductors with Si platforms. However, for the release of target epitaxial layers from III-V heterostructures, it is required to embed a mechanically or chemically weak sacrificial buffer beneath the target layers. This requirement severely limits the scope of processable materials and their epi-structures and makes the growth and layer-release process complicated. Here, we report that epitaxial layers in commonly used III-V heterostructures can be precisely released with an atomic-scale surface flatness via a buffer-free separation technique. This result shows that heteroepitaxial interfaces of a normal lattice-matched III-V heterostructure can be mechanically separated without a sacrificial buffer and the target interface for separation can be selectively determined by adjusting process conditions. This technique of selective release of epitaxial layers in III-V heterostructures will provide high fabrication flexibility in compound semiconductor technology.