The intriguing tie pits the National League leaders against the League One table-toppers.
The match, which will be broadcast on ITV, is generating plenty of excitement in the town.
The fact that Paul Cook, Gary Roberts, Tom Naylor and Michael Jacobs all have Pompey connections adds extra spice to the fixture.
The Spireites named a strong side in the last round, beating Kettering Town 5-0, so let’s take a look at how they could line-up for this one...
1. Ryan Boot - GK
He made his debut in the last round against Kettering Town, keeping a clean sheet. Harry Tyrer's back injury meant that he kept his place and played the last two league games, recording a shutout against York City. Tyrer is fit again so there is a tough decision to be made. My bet would be that Boot starts again, but Tyrer returns for the Barnet clash. Photo: Tina Jenner
2. Liam Mandeville - RB
Probably someone you might not want to risk with Barnet just around the corner, but he's so key to this team. Will he play right-back again? Let' see. Photo: Tina Jenner
3. Ash Palmer - CB
Tyrone Williams played out of his skin at Kidderminster so he certainly won't be 'dropped', but rather 'rested' for Barnet if that is the case. Bringing Palmer in doesn't weaken the defence - there isn't a lot to choose between them, in my opinion. Photo: Tina Jenner
4. Jamie Grimes - CB
The skipper was given a rare rest in the last round, but you'd think Chesterfield will need their captain for this one. Photo: Catherine Ivill