Aiming at confirming the chemical structure, herein, we report the crystal structure of Arbortristoside-A, isolated from the seeds of Nyctanthes arbor-tristis Linn., investigated by single crystal X-ray crystallographic analysis. The unambiguously established structure of Arbortristoside-A not only addresses previously reported structural flaws but also encourages its chemical, computational, and physiological studies as a lead drug candidate of pharmaceutical significance.