Despite interlayer binding energy is one of the most important material properties for graphite, there is still lacking report on its direct experimental determination. In this paper, we present a novel experimental method to directly measure the interlayer binding energy of highly oriented pyrolytic graphite (HOPG). The obtained values of the binding energy are 0.27($pm $0.02)J/m$^{2}$, which can serve as a benchmark for other theoretical and experimental works.