The 23-time grand slam victor had played herself into form through three rounds in her grand slam comeback, but she took to the main interview room Monday to announce her withdrawal just minutes before the match was about to begin on the Philippe-Chatrier court in Paris.
"I unfortunately been having some issues with my pec muscle, "she said, via the Roland Garros Twitter account".
She was also in just her third tournament of the year after giving birth to daughter Olympia in September.
"I gave up so much - time with my daughter, time with my family".
Williams was set to take on Sharapova on Court Philippe Chatrier just minutes later, but said the tapings she used during Sunday's doubles match against Andreja Klepac and Maria Jose Martinez Sanchez had not been effective in alleviating her injury.
The match was going to be among the most anticipated in the tournament, a clash between two of the world's biggest tennis stars. The first time I felt it was against Goerges in my last match.
Williams added: "The first time I felt it was against [Julia] Goerges in my last match".
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It was the first time the 36-year-old has withdrawn midstream from a Grand Slam singles event.
"I thought [doubles] was a flawless opportunity to see how I was going".
Williams then held a hastily-assembled news conference to confirm her withdrawal. Though she was playing excellent, it took a lot to get the point she's at now.
Williams admitted it was painful to walk away from a first match against Sharapova since the 2016 Australia Open, after which the Russian was given a 15-month doping ban. I'm not serving that often; I'm serving once every four games...
"I'm going to get an MRI tomorrow".
"It is hard because, like I keep saying, I have given up so much to be here".
Sharapova progresses straight through to the quarter-finals as a result of Williams' withdrawal. Still, Williams said, "It's very hard because I love playing Maria...."
"I don't have any negative feelings towards her, which again, was a little disappointing to see in that hearsay book... as a fan, I wanted to read the book and I was really excited for it to come out and I was really happy for her".