The exciting draw will see Paul Cook, Gary Roberts, Tom Naylor and Michael Jacobs all face their old club.
Pompey, managed by John Mousinho, are currently top of League One after 12 games and are unbeaten.
The Spireites beat Kettering Town 5-0 in the fourth qualifying round on Saturday, earning themselves £9,375 in prize money.
Winners of first round ties, to be played between November 3-6, get £40,000.
Chesterfield have reached the third round in the last two seasons.
Town are back in action on Saturday at home to Gateshead.
During the fixture against Oldham Athletic earlier in the season, a special guest made their first trip to the SMH Group Stadium. Pauline McAllister was joined by Gordon, a 22-month-old support dog.
Speaking about the experience, Pauline said: “We’re socialisers – not trainers. We’ve had lots of different experiences to get the dog used to it.
“We thought a football match would be really good to get used to the noise. We hadn’t got a clue on how to get in or how to get tickets but our experience was absolutely wonderful.
“We went to the wrong entrance and made a few phonecalls; somebody came down to meet us. They escorted us to our seats. Everybody was absolutely marvellous, it was really good.
“It was a really hot day and the dog was in the sun most of the time, so we had to leave early – we were a bit worried about the dog.
“I was quite nervous about going but we didn’t have any worries at all. The whole thing was lovely. I can’t thank everybody enough.”
Ash Palmer headed the Spireites into the lead midway through the second half and it’s safe to say that Gordon was celebrating along with the majority of 8,164 in attendance.
She said: “When Chesterfield scored, the dog jumped up and put his two paws on my shoulder.
“He was excited as we were. He is a Chesterfield fan definitely and he’s even wearing a blue bandana.”
Pauline was very complimentary about her experience at the SMH Group Stadium alongside Gordon.
She recommends it to others who are looking to help their service dog in different situations but says that it would depend on the needs of the dog.
“I think this was the first time they had a support dog there because not many people in Chesterfield actually have a support dog,” Pauline said.
“We were the first ones to go along to a football match. We said it was a really good experience and if anybody else wanted to do it, to go ahead.
“It depends on the needs of the dog, really. They have to be fairly settled otherwise all of the noise would upset the dog.”
In the near future, it is possible that another four-legged friend may come to a Chesterfield fixture: “We’ve got another little dog, Bertie. He’s only nine weeks old at the minute.
“It will be a while yet before he’ll be able to go but we will certainly repeat it again.”
