N,N'-Bis(5-hydroxysalicylidene)ethylenediamine is primarily used as a chelating agent in various chemical and industrial processes. It forms stable complexes with metal ions, making it valuable in metal ion detection and separation techniques. This compound is often employed in analytical chemistry for the spectrophotometric determination of trace metals, such as copper and iron, due to its strong binding affinity and distinct color changes upon complexation.

In material science, it is utilized in the synthesis of coordination polymers and metal-organic frameworks (MOFs), which have applications in gas storage, catalysis, and sensing. Additionally, its ability to bind metal ions makes it useful in wastewater treatment for the removal of heavy metals from industrial effluents.

In biochemistry, it serves as a model compound for studying enzyme-metal interactions and mimicking metalloenzyme activities. Its versatility in forming metal complexes also finds applications in the development of catalysts for organic transformations and in the preparation of advanced materials with specific electronic or magnetic properties.