A woman was airlifted to hospital and three others injured after they came off their motorbikes on the Western Freeway near Ballan.
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According to Victoria Police, it was reported the four riders, in convoy, "came off their respective motorcycles on Western Freeway about 1.50pm" on April 7.
"One of the riders was airlifted to hospital with life-threatening injuries," a police spokesperson said in a statement.
"The remaining three riders were taken to hospital with non-life-threatening injuries.
"The exact circumstances surrounding the incident are yet to be determined."
An Ambulance Victoria spokesperson confirmed a woman in her 60s was flown to the Royal Melbourne Hospital in a critical condition with an upper body injury, and three other women - two in their 50s and one in her 30s - were taken to the Ballarat Base Hospital in a stable condition with upper and lower body injuries.
Traffic was diverted through Ballan while the scene was cleaned up.
Anyone who witnessed the incident, has dashcam/CCTV footage or information is urged to contact Crime Stoppers on 1800 333 000 or make a report online at www.crimestoppersvic.com.au.
The incident follows a fatal motorcycle crash near Gordon on March 29.
A 59-year-old man from Glen Waverly died at the scene of the crash after losing control, police said.
