The determination was made using Holloway's dental records, according to a statement issued late this morning by the Aruba Public Prosecutor's Office.
Aruban Solicitor General Taco Stein said shortly after the announcement that they could determine quickly that the bone wasn't Holloway's -- the jawbone found on the beach had a wisdom tooth still attached and Holloway's dental records showed the teen had had her wisdom teeth removed.
Aruban authorities, Stein said, are checking with neighboring islands to see if the bone could be a match for any other missing persons.
An attorney for Holloway's mother, Beth Twitty, issued a stinging statement after the announcement, critizing the Aruban government.
"Beth accepts the forensic conclusions, is emotionally exhausted from the inexplicably long wait, and deeply disappointed in the time and manner in which she learned of the results," John Q. Kelly said in the statement today. "Apparently Aruban prosecutors were more sensitive to media concerns than the painful vigil of a mother."
A match was considered a long shot. In the five years since Holloway disappeared while on a school trip, there has been no physical evidence to confirm that she had been killed.
The Aruban prosecutors office said a Dutch forensics team could extract only a small amount of DNA evidence from the one tooth that was still attached to the mandible. It was, they said, "of low quality."
"The bone appeared to have been exposed to various (weather) influences and proved to be porous," the statement read.
For the time being, the forensics team has stopped working to test the DNA.
"There is still a possibility that the molar from the jawbone could be used for further efforts to generate a DNA profile," the statement read. "However, this follow-up study of said molar would inhibit future research into identifying the person from whom the bone originates."
It is believed a recent hurricane may have stirred up the beach, leading tourists to find the bone outside a hotel.
Holloway, an 18-year-old high school senior from Alabama, vanished in the island country in May 2005. The suspect in her disappearance has long been Dutch playboy Joran van der Sloot, who has never been formally charged.
Van der Sloot, who was 17 years old at the time of Holloway's disappearance, is now being held in a Peruvian prison after confessing to the murder of Stephany Flores, 21, earlier this year.
There have been many false hopes regarding Natalee's disappearance in the five years since she went missing.
Beth Twitty , Holloway's mother, has made several attempts to get information out of van der Sloot, who has promised to talk then reneged or spoken only in vague terms.
Theories, Rumors Abound in Natalee Holloway Case
Hidden-camera video footage surfaced last week of Twitty meeting with van der Sloot in the rat infested Castro Castro prison in Peru, imploring him to talk to her, even offering to help him.
"I want to know what happened and I want to move on, Joran," Twitty is seen telling van der Sloot, who was 17 years old at the time Holloway disappeared. "I want to move on in my life and I can't close the book."
Van der Sloot, who let Twitty do most of the talking during the portions of the tape that have been released, said in response that he has "never listened to anyone who's meant well for me."
"It's very hard for me to talk to you. It's really not easy," he says in the video, released as part of a Dutch documentary. "I've made so many bad decisions for all the wrong reasons. ... I'm really very addicted to perks, especially gambling. That's why I've told so many lies."
Twitty was left in tears at the end of the meeting with no sign that she had come any closer to finding her daughter.
There have been numerous theories about what happened to the pretty blonde teenager. Some have speculated that van der Sloot threw her body into the ocean, while others have claimed she was sold into the sex trade.
Aruban officials sent a dive team into the ocean in March after a Pennsylvania couple captured on camera what they said was an underwater shot of Holloway's remains. Forensic experts were skeptical and the search was called off after nothing was found.
ABC News' Jeremy Hubbard and Jenna Mucha contributed to this story.