The incident occurred between Tallarook and Seymour, and 10 passengers were on board, as well as one conductor.
After the two carriages separated, they came to a controlled stop and no injuries were sustained.
The driver of the Sprinter train was able to notify the operations centre, and the train was transported back to Seymour.
For about three hours, coaches replaced services on the line between Seymour and Southern Cross Station, transporting passengers to Craigieburn first to continue their journey towards Melbourne.
Train services resumed at about 9am.
State Member for Euroa Annabelle Cleeland issued a statement, urging the Minister for Active and Public Transport to investigate the incident.
“Today’s shocking incident where a V/Line train reportedly lost a carriage near Tallarook is a damning example of the Allan Labor Government’s failure to deliver safe and reliable public transport for regional Victorians,” Ms Cleeland said.
“It should not take a near-disaster to expose the government’s failure to invest in maintenance, safety and reliability.”
A V/Line spokesperson said the incident was under investigation.
“Safety is our top priority as we investigate the cause of a train fault on the Seymour Line this morning,” the spokesperson said.
“Trains have now resumed, and we thank passengers for their patience.”