Anhalt-Bernburg was a principality of the Holy Roman Empire and a duchy of the German Confederation ruled by the House of Ascania with its residence at Bernburg in present-day Saxony-Anhalt.

It was created in 1252, when the Principality of Anhalt of the Holy Roman Empire was partitioned among the sons of Henry I into Anhalt-Aschersleben, Anhalt-Bernburg and Anhalt-Zerbst.

When the line of Anhalt-Aschersleben became extinct in 1315, Prince Bernhard II of Anhalt-Bernburg claimed their territory, he could however not prevail against his cousin Albert, Bishop of Halberstadt.

Re-united Anhalt was again divided in 1603 among Prince Joachim Ernest's sons into the lines of Anhalt-Dessau, Anhalt-Köthen, Anhalt-Plötzkau, Anhalt-Bernburg and Anhalt-Zerbst.

In 1803 Prince Alexius Frederick Christian of Anhalt-Bernburg was elevated to the rank of a duke by Emperor Francis II of Habsburg.