On January 5, 1810, the Virginia General Assembly recognized 20 acres of land owned by farmer and trader Thomas Buffington at the confluence of the Guyandotte and Ohio rivers as the new village of Guyandotte. A map showed six streets, three running east and west intersecting with three running north and south.

By the late 1830s, Guyandotte was a frequent port of call for steamboats traveling the Ohio River and a busy stagecoach stop on the James River & Kanawha Turnpike. The village boasted 40 homes, five stores, a non-denominational church, a primary school, and a gristmill said to be the largest between Cincinnati and Pittsburgh. In 1849, Guyandotte was officially incorporated and its boundaries enlarged.

The outbreak of the Civil War saw strong support for the South in Guyandotte, with many residents leaving to fight for the Confederacy. A Union post was established in the village, which on November 10, 1861, was attacked by a 700-man Confederate cavalry unit. The Confederates easily overcame the Union forces, most of whom were raw, untrained recruits, but they withdrew the next day when fresh Union troops arrived. Angered by the collaboration between the Confederates and the local residents, the Union troops burned the village.

With rail tycoon Collis P. Huntington's establishment of his new town on the Ohio just downstream from Guyandotte in 1871, the economic fortunes of Guyandotte began a steady decline. In 1911 the citizens voted (260 to 70) to become part of Huntington.