She smashed legendary sprinter Black Caviar's Australian win record last August when making it 26 in a row, and surpassed retired northern hemisphere jumper Hurricane Fly's then-record 22 Group One victories at the Chipping Norton Stakes last month.
Winx came in wide before breezing past her challengers with 200 metres to go.
Asked how he felt, jockey Hugh Bowman said: "It's over, isn't it?"
The world's top-rated turf horse sprinted home to claim her third consecutive Queen Elizabeth Stakes over 2,000m, in front of 50,000 boisterous fans at Royal Randwick in Sydney. "She's just an wonderful horse for everybody to be part of and I'm just a small cog in the wheel".
The star mare's win in the $4 million race was her 33rd consecutive victory and the 37th of her illustrious career.
Trainer Chris Waller in his usual lip wobbling emotion after the race said Winx has that rare ability to bring people together, and not just in Australia. "Everyone watching around the world", he said.
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Bowman settled Winx in sixth place in the nine-horse field and he made his run on her at the 600-metre mark. America, Europe, Africa, all over Australia. "(Winx) has captured the hearts of people all around the world". "She is the most incredible, supreme athlete I think I've ever seen".
Adding to the moment, Winx became the 100th Group One victor for Waller.
Winx has also claimed her 20th victory at Randwick, where she has picked up the vast majority of her 33 race wins.
Winx, unlike so many human athletes, has been able to end the journey on her terms.
Such is her legend that Winx has become more than just a horse in Australia.
Legendary Australian sports broadcaster Bruce McAvaney said ahead of the race that her farewell sat comfortably alongside any of the great Australian sporting moments he had called or witnessed in his long career. "And it is such a special moment to share it with not only my attractive wife and my handsome children, but the lovely people who have been right behind Winx", he said.