Emergency services were called to the A911 from Glenrothes to Windygates, near Markinch, where the crash happened shortly before 1:30pm yesterday.
The 45-year-old woman was a passenger in an Audi A5 Coupe. Firefighters had to cut her free from the vehicle and she was airlifted to hospital in Dundee. She died around 7:25pm.
The male driver of the car had only minor injuries and was taken to hospital in Dunfermline. The lorry driver was treated by ambulance staff at the scene.
Chief inspector Davie McCulloch, of Fife Police, said: “It’s essential we get as much relevant information as possible and I’m appealing to anyone who saw either the car or the lorry prior to the collision, or the collision itself, to contact us.”
Meanwhile, a man died after his car ploughed into a block of flats at Salamander Court, in Leith, around 7pm last night.