His plea in Florida on Tuesday (Wednesday AEDT) came hours after a sheriff's report said he had pain pills and showed signs of impairment at the scene of last week's crash.
The online court docket for Martin County, Florida, showed Woods entered a written plea of not guilty and planned to waive his appearance during an arraignment hearing next month.
Earlier Woods told authorities he was looking at his phone and did not realise the truck in front of him had slowed before his rollover crash in Florida last week.
According to a probable-cause affidavit obtained on Tuesday, Woods had two hydrocodone pills in his pocket and officers observed him to be lethargic, slow, "sweating profusely" with eyes that were bloodshot, glassy and pupils that were "extremely dilated".
The report said deputies found two white pills, later identified as the opioid hydrocodone used to treat pain, in his pocket.
When asked during the criminal DUI investigation if he took any prescription medication, the report said the 50-year-old golfer replied "I take a few", adding he had done so earlier in the morning.
Woods' manager did not immediately respond when asked to comment on details of the probable-cause affidavit.
A Martin County sheriff's deputy wrote in the report that Woods, when asked about the collision, said he was looking at his phone and changing the radio station, which caused him not to see a truck slowing before the crash.
The officer said in the report he observed Woods "limping and stumbling" and added that the golfer told him he has had seven back surgeries and more than 20 operations on his leg.
Woods, a 15-time major champion and the greatest golfer of his generation, was arrested last Friday afternoon on a charge of driving under the influence after his Land Rover rolled over on a two-lane road near his Jupiter Island home, having travelled at "high speeds" and clipped the truck.
He was released on bail that night. No one was injured in the crash. According to the sheriff's report, the truck sustained $US5000 ($A7250) damage.
The officer also noted in the report that Woods was "extremely alert and talkative" and had "hiccups during the entire investigation".
Woods, who said last Tuesday he was hoping to be ready to compete in the Masters from April 9, told the officer he has a limp and that his ankle seizes while walking.
The deputy who walked Woods through a series of field sobriety tests said in the report that, based on his training: "I believed that Woods' normal faculties were impaired, and he was unable to safely operate the motor vehicle."
- with AP