An hour before game time, the Royals announced that Guillen was out with tightness in the left hip flexor, an ailment that has bothered him for several days.
Mark Teahen moved from left field to replace Guillen in right field and designated hitter Billy Butler was elevated from sixth to Guillen's fourth spot in the batting order.
Esteban German was added to the lineup, playing left field.
Center fielder David DeJesus, who had been iffy because of a stiff back, was back in the starting lineup. He did not start Wednesday's game at Oakland but entered the game on defense in the last inning.
Dick Kaegel is a reporter for MLB.com. This story was not subject to the approval of Major League Baseball or its clubs.