A 2024 study in the American Economic Review found that this funding significantly improved the medical sector, improved physician quality and fostered medical innovation, in addition to reducing infant mortality and long-run mortality.

In the Indenture of Trust, Duke specified that he wanted the endowment to support Duke University, Davidson College, Furman University, Johnson C. Smith University; non-profit hospitals and children's homes in the two Carolinas; and rural United Methodist churches in North Carolina, retired pastors, and their surviving families.

[4] The Duke Endowment is a co-publisher, along with the North Carolina Institute of Medicine, of the North Carolina Medical Journal, a journal of health policy analysis and debate.

The remaining funds were to be invested (primarily in Duke Power Company stock).

Of the income generated annually by these funds, 20% were to be reinvested, each trustee was to receive 0.2%, and the rest was to be paid out in the following allocation: