Holliday pulled up just before reaching first base while running out a grounder, which was the second-to-last out of the Rockies' 9-2 loss to the Mets at Coors Field on Saturday.
Trainers immediately iced the injured area, and Holliday said after the game that his hamstring was too numb for him to have any idea how badly he was hurt. The answer came Sunday morning. Holliday will not be with the club as it heads to Philadelphia to start a 10-game road trip.
"We're going to leave Matt back for a few days because we have all the pertinent training rehab stuff here," manager Clint Hurdle said. "We'll revisit when he might catch up with us."
The Rockies placed shortstop Clint Barmes (right knee sprain), who was leading the team with a .343 batting average, and right fielder Brad Hawpe (strained right hamstring) on the DL on Saturday morning.
Holliday's injury is yet another blow to a club that won the National League title last year but is underperforming, with a 19-30 record going into Sunday.
Holliday, a two-time All-Star who finished second in National League Most Valuable Player voting last season, was hitting .321 with eight home runs and 26 RBIs. Holliday homered in the first inning on Saturday to extend his hit streak to 10 games.
Original starting shortstop Troy Tulowitzki (torn left quadriceps tendon) was injured April 29, long reliever Kip Wells (blood clots in the right hand), right-handed setup man Luis Vizcaino (shoulder) and bullpen left-hander Micah Bowie (strained forearm) have been out for some time as well. In addition, right-handed starter Jason Hirsh (strained shoulder) hasn't pitched all season.
And the Rockies went into Sunday's game against the Mets without third baseman Garrett Atkins (stiff neck), who was hitting .338, led the team with 30 RBIs and was tied with Holliday for the club lead in homers. The Rockies also were hoping that center fielder Willy Taveras (bruised right knee) would be available for late-game duty after having to sit out Saturday's game.
The Rockies called up Cory Sullivan from Triple-A Colorado Springs. Ryan Spilborghs replaced Holliday in left while Sullivan played a reserve role.
Sunday's starting lineup included four players who began the season in Colorado Springs -- second baseman Jonathan Herrera, right fielder Seth Smith, third baseman Ian Stewart and shortstop Omar Quintanilla.
Thomas Harding is a reporter for MLB.com. This story was not subject to the approval of Major League Baseball or its clubs.