Accurate measurement of interfacial properties is critical any time two materials are bonded-in composites, tooth crowns, or when biomaterials are attached to the human body. Yet, in spite of this importance, reliable methods to measure interfacial properties between dissimilar materials remain elusive. Here we present an experimental approach to quantify the interfacial fracture energy Γ i that also provides unique mechanistic insight into the interfacial debonding mechanism at the nanoscale. This approach involves deposition of an additional chromium layer (superlayer) onto a bonded system, where interface debonding is initiated by the residual tensile stress in the superlayer, and where the interface can be separated in a controlled manner and captured in situ. Contrary to earlier methods, our approach allows the entire bonded system to remain in an elastic range during the debonding process, such that Γ i can be measured accurately. We validate the method by showing that moisture has a degrading effect on the bonding between epoxy and silica, a technologically important interface. Combining in situ through scanning electron microscope images with molecular simulation, we find that the interfacial debonding mechanism is hierarchical in nature, which is initiated by the detachment of polymer chains, and that the three-dimensional covalent network of the epoxy-based polymer may directly influence water accumulation, leading to the reduction of Γ i under presence of moisture. The results may enable us to design more durable concrete composites that could be used to innovate transportation systems, create more durable buildings and bridges, and build resilient infrastructure. molecular mechanics | bimaterial systems | superlayer | energy release | biomedical I nterfaces exist whenever materials are bonded together and can be found frequently in both natural and synthetic bonded systems, for instance, the mineral-protein interfaces in animals, interfaces between different phases in composite materials, the enamel-polymer interfaces involved in dental treatment, or the cell-substrate interfaces in biomedical applications (1−7). The integrity of the interface under various environmental conditionsincluding different temperature and moisture levels-is critical to many applications (8−10). Meanwhile, because of the advancement and development in technology, a large number of bonded systems in various engineering applications are needed, possessing increasingly higher accuracy in design and manufacturing process in a very small length scale. However, currently, a robust and generally applicable methodology to quantify the fracture energy at these interfaces from a microscopic perspective is lacking.Several straightforward measurement methods for quantifying the interfacial fracture energy on large specimens, such as direct peel/shear specimen (11, 12), Brazilian disk specimen (13), and sandwiched beam specimen (14, 15), have been reported. However, it has been difficult to directly measure this parameter for microscale...