The video, seen by Al Jazeera and released over the past week, is the second installment of ISIL footage to feature the captured Kurdish fighters.
In the new video, the Kurds, dressed in orange jumpsuits, their heads bowed, are paraded through an enclosed area and then seen in individual steel cages.
It comes just days after a first video showed 17 Kurdish fighters been taken on a procession through the streets of Iraqi town of Kirkuk.
ISIL fighters spoke to the camera in Kurdish, reiterating "our war is not with the Kurdish Muslim people but with the infidels and their treacherous agents".
Halgord Hikmat, media adviser to the ministry of Peshmarga, confirmed to Al Jazeera the authencity of the video.
"Officially we don't have any negotiation with them because we know they do respect international human rights, or any law war or prisons," Hikmat said.
On Tuesday, Kurdish President Masoud Barzan visited Kirkuk for the first time since ISIL's assault on the town.
Barzan said the Kurdistan Regional Government will spare no effort to free Peshmerga hostages from ISIL.
"If ISIL decides to kill them, they will pay a heavy price," Barzani said.