The Seasiders will welcome Darlington to the Towbar Express Stadium at the Turnbull Ground on Saturday, 7th August, with kick-off set for 3pm.
It will be the first time since 2016 that the two sides have faced each other, with the Quakers running out 7-1 winners on that occasion as they secured the Northern Premier League Premier Division title.
The game against Alun Armstrong's men, who currently play in the National League North, is the second fixture announced by Whitby ahead of the 2021/22 campaign.
Lee Bullock and Nathan Haslam's side will face York City on Saturday, 3rd July, following on from the Whitby Town Community Cup the previous week.
Ticketing information for the game against Darlo, along with the Community Cup and the York friendly, will be confirmed on whitbytownfc.com in due course, along with details of any further pre-season matches.