Notts County were beaten 2-0 by Newcastle United's Under 21s in the second match of their Northern Group G Checkatrade Trophy campaign.
Goals from Cal Roberts and Elias Sorensen won the game for the Tynesiders on an evening when Tyreece Kennedy-Williams and Kion Etete were given their senior debuts for Notts.
Doncaster's victory over Grimsby means Notts could still qualify for the knockouts if they beat Rovers in the final match of the group stage in November.
Harry Kewell made 10 changes to the side that beat Macclesfield on Saturday.
Shaun Brisley was the only man to keep his place, this time lining up in a more familiar role alongside Dan Jones in central defence.
Kennedy-Williams and the more experienced Andy Kellett were given the full-back slots, with Nathan Thomas and Sam Osborne ahead of them.
David Vaughan linked up with Noor Husin in central midfield and Will Patching supported Kristian Dennis up front.
The visitors, kicking towards the Air-IT Kop in the first half, were handed a golden opportunity to make the dream start when Kellett bundled into Tom Allan in the area, conceding a penalty.
Cal Roberts fired the ball straight down the middle and, while Branislav Pindroch's outstretched foot made contact, it hit the back of the net.
Notts' response came from down the right, Osborne's cross being met by Thomas who saw his header saved on the goalline by Nathan Harker.
Vaughan fired a long-range effort wide and Patching bent a free-kick over the bar as the hosts continued their pursuit of an equaliser.
And it nearly came on 30 minutes when Thomas' free-kick looked destined for the top corner, only for Harker to push the ball over.
Sorensen let a good headed opportunity pass him by before Dennis' effort from range whistled wide.
And it was left to Roberts to claim the final chance of the half, the winger sending a sweetly-struck effort over the bar before the referee called time on the first half.
There were no Notts changes at the break and nine minutes after the restart, Vaughan wrote his name into the book for a foul on Kelland Watts.
Patching saw an effort diverted behind for a corner moments later - but it was to be the visitors who struck next.
Roberts turned provider this time, playing a delightful through ball for Sorensen who poked his shot beyond Pindroch with an accomplished finish.
Kewell made his first change just after the hour, sending Remaye Campbell on for Patching shortly before Allan saw an effort diverted just wide of Pindroch's post by Jones.
Osborne was the next player to have his name taken, for a late tackle on Matty Longstaff, before Etete replaced Kennedy-Williams for his senior debut.
The youngster lined up alongside Dennis and Campbell as Kewell switched to a 4-3-3, with Osborne filling in at right back.
Brisley headed a Jones cross wide, Osborne fired an effort off-target and, at the other end, Roberts missed a gilt-edged chance at the back post as the clock ticked down towards full-time.
There was still time for young midfielder Alex Howes to replace Vaughan before six minutes of added time were indicated by the fourth official.
But Notts were unable to mount a recovery as the game ended 2-0.