The Spireites and Swindon Town were separated by a second-half penalty to give the home side a 1-0 win at the Nigel Eady County Ground.
Four changes were made following last Saturday’s defeat to Walsall – Ryan Boot, Janoi Donacien, Jenson Metcalfe and Liam Mandeville were all recalled to the starting XI.
After a first half that was marred by a long stoppage after Tyrone Williams collided with Boot, Kabongo Tshimanga scored the only goal of the game to give Swindon the victory.
The first major incident came in the ninth minute. Harry Smith was booked after an incident off-the-ball with Jack Sparkes. Swindon manager Ian Holloway was then shown a red card for his protestations.
Five minutes later, Williams collided with Boot and had to be taken off the pitch on a stretcher. He was replaced by Tom Naylor.
After the restart, Smith took aim and saw his shot deflected and claimed by Boot.
Michael Olakigbe tried the spectacular with a 30-yard half-volley, but his shot was caught by Connor Ripley.
Chesterfield looked the more dangerous but struggled to create the golden opening. Dylan Duffy’s powerfully low-hit free-kick just before the break was kept out by the Swindon goalkeeper.
The home side, who struggled to make their way into the Chesterfield area in the first period, started the second period with a higher tempo.
Swindon threatened from three separate corner kicks – one was headed wide by Miguel Freckleton, Metcalfe blocked another, and Smith nodded over from the third.
Soon after, the hosts were awarded a penalty as Boot caught former Spireite Tshimanga in the penalty area. The striker dusted himself off and calmly tucked away the resulting spot kick.
Ollie Banks, Paddy Madden, Ryheem Sheckleford and Kane Drummond were all brought on in an attempt to find an equaliser.
With ten minutes of normal time to go, Sparkes’ corner bounced off the midriff of Bim Pepple and the ball went just wide of the post.
The Blues unfortunately couldn’t find the elusive equaliser and had to settle for a second defeat in a row.
Chesterfield: Boot, Donacien (Sheckleford 76), Williams (Naylor 25), Grimes ©️, Sparkes, Metcalfe. Olakigbe (Drummond 76), Mandeville (Banks 69), Dobra, Duffy (Madden 69), Pepple.
Unused substitutes: Thompson, Oldaker.