The FA last year announced they will play a lead role in testing the idea - proposed by football's law-making-body the International Football Association Board (IFAB) - which will allow teams to make a fourth change in extra time.
The scheme will be trialled for the quarter finals, semi-finals and final of this season's FA Cup meaning Arsenal, Chelsea, Millwall and Tottenham will be able to benefit from it for their sixth-round ties.
The Gunners will host non-league Lincoln City at Emirates Stadium on 11 March in the late Saturday kick-off which follows on from Middlesbrough's quarter-final game with either Huddersfield or Manchester City.
Spurs boss Mauricio Pochettino could also opt to use a fourth sub in extra-time when Tottenham take on Millwall at White Hart Lane a day later.
Chelsea then host Jose Mourinho's Manchester United side in their sixth-round tie at Stamford Bridge on Monday night.
The idea was first trialled at last summer's Copa America, and comes after the FA decided to give the latter stages of the FA Cup, from the quarter-final round onward, a straight knockout format.
Speaking about the fourth substitution trial, FA chief executive Martin Glenn said: "The Emirates FA Cup is renowned for its great history and tradition but, as the game’s governing body, it is important The FA continues to look at how it can further add to the drama and spectacle of a competition loved by millions around the world.
"With The Cup now adopting a straight knockout format from the quarter finals onwards, the introduction of a fourth substitute in extra time will bring extra intrigue and interest.
"Also, from a technical point of view, it will be interesting to see how managers use the chance to make an additional substitution in such high-profile games and the impact it has on the final result.
"Player welfare and being mindful of the number of games people play at the elite level has also been a consideration."
