Van der Sloot, who denies any wrongdoing, appeared mostly relaxed throughout the TV appearance, despite remarks from crime reporter Peter R. De Vries challenging his honesty.
At the end of the Pauw & Witteman program, Van der Sloot and De Vries had a testy exchange in which Van der Sloot questioned whether De Vries would "be man enough" to apologize if evidence exonerating him came to light.
The live show ended with guests shaking each others' hands. Van der Sloot stood up, then abruptly grabbed a glass of wine and leaned across the table to toss the contents directly in De Vries' eyes at a moment when the reporter was looking down. That segment was broadcast Saturday night by a different news program, Nova.
Van der Sloot initially told investigators he last saw Holloway when he dropped her off at her hotel, but later he said he left her on the beach near her hotel. He has acknowledged the initial lie, and said he regrets it.
Holloway disappeared on May 30, 2005, hours before she was to return home to Mountain Brook, Ala. She was on the final night of her school graduation trip to the island.
Van der Sloot and two other former suspects, brothers Deepak and Satish Kalpoe, were seen leaving a bar with her that night. They were questioned early in the investigation and again late last year before being released. All three deny involvement in Holloway's disappearance.
Van der Sloot's Dutch lawyer and the show's producers could not immediately be reached for comment.
"He did it out of a kind of frustration, annoyance," De Vries said on Nova Saturday. "I spoke with him later, and he apologized. And then he said that he actually wanted to curse me out during the broadcast, but he was able to refrain. But at the end, before he knew it, he had thrown the glass of wine in my face."
Show host Paul Witteman said Van der Sloot had told him afterward he had planned the act during the show.
"The further along the show went, the more he was annoyed because he had to explain again and again why he had lied" at the start of the investigation into Holloway's disappearance, Witteman said Van der Sloot told him.
Van der Sloot thought, 'Now I'll grab my chance, now I can do it' (throw the wine), and then he did," Witteman said.
Witteman said he thought the move would damage Van der Sloot's credibility, the opposite of what his family had hoped the effect of appearing on the program would be.
Prosecutors say they cannot prove a crime was committed without a body, but they believe Holloway is dead.