Police confirmed speed and alcohol are not believed to be factors in this crash, however investigations are continuing.
Last night, police initially said the man was being airlifted to hospital by rescue helicopter in a critical condition, but a short time later they advised he had died.
The highway was closed between Alexandra and Clyde and diversions were in place through Airport Rd, Dunstan Rd and Springvale Rd following the incident.
Alexandra Police are still keen to hear from anyone else who may have information about the crash, on (03) 440 2500.