The run is five kilometres in length and members of the public are invited to enter.
Approximately 500 people are expected to join in the fun, taking advantage of the unique opportunity to run along a real runway at daybreak.
Registration on the day will commence at 5am with a warm up at 5.40am, the run will begin at 6am and will finish by 7am so that the runway can be cleared ready for commercial flights of the day to begin.
Runners will need to register beforehand and the entry cost is £20 with all proceeds going to the airport Charity of the Year, Hampshire and Isle of Wight Air Ambulance (HIOWAA).
Dave Lees, managing director of Southampton Airport, says: ‘We are very excited to open our runway up to the public once again.
"This year we are hoping to get even more people involved and even more money raised with the goal of hitting the £10,000 mark to celebrate the Hampshire and Isle of Wight Air Ambulance 10th Birthday.
"This is an incredibly worthwhile cause and we hope to see many people taking part, despite the early start!"
Ray Southam, corporate fundraiser at HIOWAA, says: "The Runway Run is a fantastic idea and a unique opportunity that will raise much needed funds to keep the Air Ambulance flying and saving lives.
"We are very grateful to Southampton Airport for all of the effort that they have put into making it happen and for allowing us to make use of their runway. I look forward to dusting off my trainers and joining in the fun!"
A range of prizes and trophies will be given out on the day for categories such as fastest runner and best dressed.
For more information and to register for the Southampton Airport Runway Run please visit www.southamptonairport.com/run