Barry Ferguson, Lee McCulloch and Christian Dailly, all of Rangers, have withdrawn from the Scotland squad for Wednesday's friendly against Croatia.
Ferguson saw a specialist in England last week with an ankle problem which has been troubling him for several weeks.
He was able to play in the Premier League win over Hibernian on Saturday after taking an injection but has been ruled out for the game at Hampden which will be the first in charge for new coach George Burley.
McCulloch also misses out after a thigh injury flared up at the weekend, while an X-ray has revealed Dailly is suffering from a broken toe.
All three will receive treatment during the week and are expected to be fit in time for the Old Firm derby against Celtic next weekend, according to the club`s official website.
Rangers goalkeeper Allan McGregor and striker Kris Boyd remain in the national team squad.