Worthington was born on March 4, 1806, in Colchester, Connecticut, to Daniel Worthington, Sr., and Elizabeth "Betsey" Hazard. His siblings were Robert Worthington I, Daniel Worthington, Jr., Joel Worthington, Samuel Worthington, Eliza Worthington Crawford, Dr. Robert Hazard Worthington II, Giles Worthington, and Ophelia Worthington.^[1]

After living for a time in Albany, New York, Worthington moved to Oconomowoc, Wisconsin.^[2] He married Martha Searing on Christmas Eve of 1829. He had three children: Robert Searing Worthington, Hattie Worthington, and William Henry Worthington I. Martha died in 1839. The following year, on June 3, 1840, he married Mary Ann LaGrange, with whom he had seven children: Denison LaGrange Worthington, William Henry Worthington II, Mary Frances Worthington Hennegan, Martha Worthington, James LaGrange Worthington, Gerrit Hazard Worthington, and Frank Tows Worthington. Mary Ann died in 1856. In 1860, he married Julia McNaughton Proudfit, the sister of Andrew Proudfit. Worthington died on April 23, 1880, in Denver, Colorado.

Worthington was first involved in the grocery business in 1835. He owned Worthington and Davis wholesale grocery stores until his move to the Wisconsin Territory in 1847. While in Albany, he served on the city council. After moving to Wisconsin, he was involved in farming. He was elected to the Wisconsin Assembly in 1852 and was re-elected in 1854. He was elected to three two-year terms as a member of the Senate, first from 1855 to 1856, and then again from 1858 through 1861. In 1859, he was elected president pro tempore of the Senate. He was secretary of Madison Mutual Insurance Company from 1861 to 1874, when resigned due to poor health. He then moved to Denver, Colorado, where he spent his final years.