Libya has not been serviced by any international ferries for 25 years.
Travel links with and within the country were severely disrupted by the 2011 revolt that overthrew longtime dictator Muammar Qaddafi, and years of unrest which followed.
During Qaddafi's rule, from 1969, the country was under sanctions and largely closed to tourists.
The revived crossing to Turkey is operated by Libyan company Kevalay. It should take 48 hours and the vessel is set to return on December 7.
"We have deals to launch other links, including with Egypt and Tunisia," said Taha Hadid, an official at the port of Misrata, Libya's third city.
Misrata, about 200 kilometres (125 miles) east of Tripoli, is home to numerous Turkish businesses.
Air travel has also begun to return.
Airports were heavily damaged in Libya's fighting and many air links were suspended, but some have resumed since an October 2020 ceasefire between the main rival camps.