Soren Kragh Andersen, who showed signs of concussion after he fell down a hillside in a mass crash on Friday, quit the Tour de France before Saturday's stage.
Frenchman Warren Barguil of Arkea-Samsic, who was caught in the same mass crash, dropped out earlier on Saturday morning before the start of stage 14 in Carcassonne.
Kragh Andersen, who won two stages on last year's Tour, was caught in a pile up involving more than 20 other riders on a gravel stretch on stage 13 some 65km from the finish in Carcassonne.
"We closely monitored Soren during the evening and very quickly came to the decision that he was no longer fit to race after seeing some signs of concussion," said DSM team doctor Camiel Aldershof in a statement.
As riders piled up on the outside of a corner, some toppled over the edge and into a deep, wooded gully.
Kragh Andersen was the last one to climb out.
"I am really disappointed and don't want to leave the Tour de France," he said in the DSM statement.
"In my heart I know that the decision of the team's medical staff can only be the right one."
The crash had already ended the Tour for several riders.
Germany's Roger Kluge (Lotto), Briton Simon Yates and Australian Lucas Hamilton (BikeExchange) all retired on Friday.
The two withdrawals on Saturday morning leave the Tour peloton at 149 out of 184 riders who started in Brest a fortinght ago.
The Tour has been punctuated by crashes. The professional riders association staged a protest before stage four "to show their dissatisfaction with safety measures".
"I hope this is the last of the bad luck for the guys here at the race," Kragh Andersen said in his statement.