The astronaut from Westbourne, near Chichester, personally addressed the Queen in a video message live from the International Space Station (ISS) after she sent him her own well-wishes.
In the minute-long video, Major Peake, who was launched into space last month, thanked the Queen and said he hoped his journey would serve as a symbol of “international collaboration” and help make the world a better place.
The “humbled” astronaut concluded his message with what he believed was the first “God save the Queen” uttered from space.
Floating in front of a camera with a microphone, Maj Peake said: “I’m truly honoured to receive Your Majesty’s message, inspired by its wording, and humbled to be able to represent the UK and Europe on board the ISS. I can tell you it’s incredible to be able to look at the British Isles for the first time from space.
“Our islands and our nation are a special place to me and I hope the next six months will demonstrate to everyone across the UK and Europe how international collaboration and brave endeavour can change our world for the better.
“So thank you again for your well wishes, ma’am. I hope I can bring the country together to celebrate Britain in space and our great tradition of scientific exploration.
“And before I sign off, I’m not sure that this has ever been said in space before, so I’ll be the first: God save the Queen.”
A message sent by the Queen to Maj Peake shortly after his launch in December read: “Prince Philip and I are pleased to transmit our best wishes to Major Timothy Peake as he joins the International Space Station in orbit.
“We hope that Major Peake’s work on the Space Station will serve as an inspiration to a new generation of scientists and engineers.”