The youth team came from behind to beat Burton Albion 2-1 at St George's Park on Tuesday afternoon.
The hosts took the lead against the run of play before the young Spireites equalised through Javanne Smith's deflected effort from 20 yards to make it 1-1 at the break.
Chesterfield had further chances to score before Callum Brook scored the winner with a calm finish following a corner.
Reflecting on the game, coach Neil Cluxton said: "Where previously we've been deflated after going behind, we just seemed to go again and scored a great goal.
"We had a positive discussion at half time and the lads just pushed on in the second half.
"We've had a difficult start to the season so we've had a couple of meetings over the last ten days where we've addressed a few issues and there have been a few tactical changes.
"Virtually everybody is back fit now and being able to pick from a full squad makes a difference."
Spireites: England, Tansley, Gibson, Geeves, Brook, Wilkinson, Emery, Fuertado, Papworth, Smith, Netherwood
Subs: Wharton, Clogg, Wild, Turner, Hargreaves