The crash on Wednesday near the town of Urda, 130km south of Madrid, has left 69 people dead and almost 200 people injured.
No further victims were expected to be found, regional authorities said.
Consular assistance was being provided to the family of two Australians in connection with the crash, the Department of Foreign Affairs and Trade said in a statement on Friday.
"Our heartfelt condolences are with the families and loved ones of those killed, and with those injured and all affected by the train collision in Spain," a spokesperson said.
Government frontbencher Jason Clare on Thursday said consular officials were on standby to help any affected Australians or their families.
"The whole country is in shock at the moment, seeing these images on our screens. It's just horrific," he told ABC TV.
More than 500 people were aboard when a train operated by state-owned rail company Renfe crashed into the back of a stationary train operated by Iryo, a private company.
Spanish authorities are under pressure to ascertain the cause of the crash after audio recordings of interactions between train drivers and a control centre were published by the media.
Investigators are focusing on the failure of the automatic train protection system and potential errors made by the rail control centre in Madrid.
"We are working day and night not only to support the victims' families, but also to establish every last detail of this tragedy and strengthen safety measures to ensure it never happens again," Spanish Prime Minister Pedro Sanchez said.
Mr Clare on Thursday said the crash reminded him of Sydney's Granville train disaster, in which 83 people died and 213 were injured.
The January 1977 incident occurred when a peak-hour commuter train, travelling from the Blue Mountains, derailed.
A road bridge collapsed onto two of the carriages.
"Every year we get back together at the same site, and I meet people who were in that awful disaster, whose lives were changed and transformed," he said.
"This is something people never, ever recover from."
with Reuters