Born in Albemarle County in the Virginia Colony, Hopkins was educated by private tutors.

Hopkins was appointed chief justice of the first court of criminal common law and chancery jurisdiction in 1799 and served until his resignation in 1801.

In 1812, Hopkins was appointed Commander in Chief of the western frontier (Illinois and Indiana Territory), with the rank of major general.

He participated in the Peoria War and was the commander at Spur's Defeat; after these series of losses, he resigned from active duty.

Samuel Hopkins retired to his country estate, Spring Garden, near Henderson, Kentucky, and died there September 16, 1819.