Sheffield Wednesday's Scottish international pair Steven Fletcher and Barry Bannan suffered a demoralising night as Gordon Strachan's men lost 3-0 to Slovakia in Trnava.
Bannan had kept his place in the team from Saturday's 1-1 draw with Lithuania at Hampden, while Fletcher stepped up from the bench having not played a part in that particular disappointing result.
However neither could prevent Scotland from falling to their first defeat in the World Cup qualifiers, a result which heaps pressure on boss Strachan following that draw at the weekend.
Fletcher had missed an early Scotland chance with a header but was involved in Slovakia's opener as the Owls striker looked to have been fouled in the build up to Zenit St Petersburg winger Robert Mak scoring the first of his two goals.
Adam Nemec registered the other for the home side, securing a victory and putting Slovakia level on four points with Scotland.
Fletcher was later replaced in the 76th minute by Hibernian's John McGinn.
The Scots next game, in November, is against England, who drew 0-0 in Slovenia, at Wembley on November 11.
Strachan said after the defeat: "I keep repeating, these guys don't get paid for this, they come along here because they want to be here.
"We try our best to send them back mentally sound but when you get body blows like that it can be very hard.
"I was pleased with a lot in the first-half, thinking 'I like this', and then the decision goes against us and at that point I thought it is not fair against these guys that are working so hard.
"They didn't let it affect them, still brave on the ball, still trying to make chances.
"At half-time the thing was just to keep going, use the (foul on Fletcher by defender Jan Durica being ignored by referee Martin Strombergsson) decision as a power, anger, whatever you want to call it, not to be emotional but to use it to your best.
"I didn't see the second goal but when it came, they went from being a bit laborious to being strong and it was uphill."