The Spireites fell to a 2-1 defeat to Yeovil Town at the Proact Stadium despite Curtis Weston's second-half wonder goal.
Manager John Sheridan made no changes to the squad after the defeat against Bromley at the weekend. The game got underway 15 minutes later than scheduled following a pitch inspection.
The away side took the lead after just two minutes when Myles Hippolyte rolled the ball past Luke Coddington with an effort from just inside the box.
Unfortunately, things went from bad to worse when Matt Tootle was taken off on a stretcher after eight minutes. The on-loan right-back was replaced by David Buchanan.
Yeovil were awarded a penalty in the 23rd minute after Anthony Gerrard brought down Rhys Murphy in the box. The striker took the spot-kick himself and placed the ball in the right-hand side of the goal.
Chesterfield almost clawed one back as the half drew to an end. Weston found Scott Boden with a cross but the striker’s side-footed volley whistled just wide of the post.
A change was made early in the second half as Scott Boden made way for Tom Denton.
Just after the hour-mark, Chesterfield halved the deficit when the ball fell to Weston 25 yards out and the midfielder unleashed an unstoppable volley into the back of the net past a helpless Stuart Nelson.
The Spireites were reduced to ten men not too long after the goal when Coddington was had to go off after suffering an injury. With all three substitutes having been used, Will Evans took up goalkeeping duties.
Evans proved to be a suitable replacement in goal and made a couple of saves during his stint in goal.
However, despite the nine minutes of added time, the ten men couldn’t find a way back into the game and slipped to defeat.
Chesterfield: Coddington, Tootle (Buchanan 8), Evans, Gerrard, Hollis, Sheridan, Maguire, Weston, Gevaro (McGlashan 59), Boden (Denton 49), Fondop.
Unused Substitutes: Jalal, Shaw.
Attendance: 3,161 (94 away)