Born into the Brandenburg-Ansbach line of the House of Hohenzollern, Dorothea Charlotte was the seventh child and elder daughter of the Albert II, Margrave of Brandenburg-Ansbach (1620–1667), by his second marriage to Countess Sophia Margaret of Oettingen-Oettingen (1634–1664), daughter of Joachim Ernest of Oettingen-Oettingen (1612-1659) and his first wife, Countess Anna Sibilla von Solms-Sonnenwalde (1615–1635).

[1] On 1 December 1687 she married Ernest Louis, Landgrave of Hesse-Darmstadt, as his first wife.

At the time of their marriage, he was still under the guardianship of his mother, Elisabeth Dorothea of Saxe-Gotha-Altenburg until 1688, when Ernst Louis came of age.

In cooperation with Philipp Jakob Spener, whose patron she became, she promoted pietism at the court and the local University.

Ernst Louis and Dorothea Charlotte had five surviving children, two sons and three daughters.