Anhalt-Plotzkau is created for a second time in 1603 with the partition of the unificated principality of Anhalt; this time, in order to created a bigger principality, parts of Anhalt-Bernburg are extracted.

This second incarnation lasts until 1665 at which point Prince Lebrecht succeeds as Prince of Anhalt-Köthen and Plötzkau returns to the principality of Anhalt-Bernburg.