Aiden McGeady and Alex Baptiste were handed their PNE debuts after joining on season-long loans on deadline-day, with Marnick Vermijl making his second debut for the club after his £500,000 switch from Sheffield Wednesday.
Eoin Doyle also came into the starting XI, replacing the injured Jermaine Beckford up front.
Also going out of the side beaten by Ipswich a fortnight ago were Bailey Wright, Chris Humphrey and Daniel Johnson.
PNE: Lindegaard, Baptiste, Clarke, Cunningham, Vermijl, Welsh, Gallagher, Spurr, McGeady, Robinson, Doyle. Subs: Browne, Makienok, Johnson, Pringle, Huntington, Hugill, Maxwell.
Barnsley: Davies, Yiadom, Roberts, MacDonald, White, Kent, Hourihane, Scowem, Hammill, Watkins, Winnall. Subs: Lee, Jackson, Kpekawa, Bradshaw, Morsy, Armstrong, Townsend.
Referee: Geoff Eltringham (County Durham)