While all eyes were on the fabulous fashion on display from the likes of Princess Beatrice and Zara Tindall, there were also some very animated reactions in the royal box.
There were cries of frustration from Queen Camilla as she and the King missed out on a winner in the King George V Stakes.
MORE FROM ROYAL ASCOT
But Princess Eugenie appeared to be victorious on Wednesday as she jumped for joy.
See all of the best reactions from race day in the gallery below…
The King and Queen's hopes of a second Royal Ascot winner were dashed when their racehorse was well beaten on Ladies' Day.
Charles and Camilla's thoroughbred Gilded Water ran in the King George V Stakes, the same race that gave them their first victory at the famous Berkshire meet last year.
The Queen kept a keen eye on the race through a pair of binoculars as the King chatted to the couple's racing adviser John Warren, and she bobbed up and down in anticipation when the horse was among the leaders.
But the couple watched as it was left behind and finished well down in the pack as Going the Distance crossed the line first.
Princess Eugenie appeared to be quids in as she jumped up and down and threw her arms in the air with joy as she was joined by her husband, Jack Brooksbank, and William and Kate's closest friends on the second day - including Lady Laura Meade.
One race had Carole Middleton cheering from her box as she watched in anticipation from the royal box with husband, Michael, and son-in-law, Prince William.
Cameras captured a private joke between Prince William and his cousins, Princess Beatrice and Princess Eugenie, as Eugenie wiggled the tassel on her hat.
The Duke and Duchess of Edinburgh shared a hearty laugh as they presented the trophy to the winner of the Ribblesdale Stakes.
LISTEN: A Right Royal Ascot
Sign up to HELLO! Daily for all the latest and best royal coverage
By entering your details, you are agreeing to HELLO! Magazine User Data Protection Policy. You can unsubscribe at any time. For more information please click here.