'The Famous Grouse Pre-Season Challenge', in association with Perth & Kinross Council, will be held at Perthshire RFC's North Inch ground on Saturday 18 August (ko 2.00pm).
The game will take place two weeks before the new Guinness PRO14 season gets underway.
A 5,000-capacity arena will be constructed close to the city centre and the event is being sponsored by The Famous Grouse.
The game will be part of a huge weekend of activity in the city, with the rugby arena also being used for other events.
Aviva Premiership champions in 2012, Harlequins have a squad packed full of international players, such as England's Danny Care, British & Irish Lion Kyle Sinckler and former Scotland winger Tim Visser.
The London club last played the Warriors in 2016 when Harlequins edged a pre-season victory 22-17 at the Twickenham Stoop.
It has also been confirmed that seven days later (Saturday 24 August) the Warriors will travel to play Northampton Saints at Franklin's Gardens to complete their pre-season preparations.
Glasgow Warriors Head Coach, Dave Rennie, said: "It's great that we're taking a game to Perth and have the chance to engage with supporters from across our region.
"We represent the West and North of Scotland, so we're excited to get on the road again after the success of our game against Northampton in Stirling last year.
"Harlequins are a top side full of international players and will provide us with the type of challenge we need as we gear up for the new season."
Leader of Perth & Kinross Council, Councillor Murray Lyle, said: "It is a great show of faith in Perth that Glasgow Warriors have chosen our city to play host to this high profile pre-season match.
"The North Inch is a fantastic location and it will be an excellent opportunity to see a professional rugby match in the heart of the city."
John Laurie, The Famous Grouse Experience, General Manager, said: "We're proud to be backing Glasgow Warriors in their pre-season preparations as they welcome one of the biggest names in English professional rugby to Perth, our home since 1896.
"We are excited to continue our long-standing backing of rugby in Scotland. This partnership gives us a chance to bring a new and unique experience to fans of the game."
CEO of Perthshire Rugby, Allan Brown, added: "Perthshire Rugby are delighted that Glasgow Warriors have chosen the North Inch in Perth as the venue for this fixture and we welcome this high profile fixture to Perth as we embark on our 150th anniversary celebrations.
"We look forward to welcoming supporters from Glasgow and all over the Caledonia region to the North Inch, the home of Perthshire Rugby and we look forward to working with Glasgow Warriors and Perth & Kinross Council to make this a tremendous success."
Glasgow Warriors Season Ticket Members who renew for the 2018/19 season, when memberships go on sale soon, will get first access to tickets in a pre-sale window from 10am on Friday 8 June until 5pm on Friday 15 June. Tickets for this game will then go on public sale from glasgowwarriors.org from 10am on Monday 18 June.