World Central Kitchen (WCK) founder José Andrés has accused Israeli forces in Gaza of targeting his aid workers "systematically, car by car".
Monday's strike which killed seven members of his staff was not a mistake, he said, repeating that Israeli forces had been told of their movements.
WCK workers from Australia, Canada, Poland, the UK and the US were killed as well as their Palestinian colleague.
Israel says the strike was a "grave mistake" and has apologised.
It has also promised an independent investigation.
According to the charity, the aid convoy was hit while leaving the Deir al-Balah warehouse, "where the team had unloaded more than 100 tonnes of humanitarian food aid brought to Gaza on the maritime route".
The convoy was made up of three vehicles, including two that were armoured, which clearly displayed the charity's logo. All three were hit during the strike.
Speaking to Reuters news agency on Wednesday, the Spanish-American celebrity chef said this was not a "bad luck situation where, 'oops,' we dropped the bomb in the wrong place".
In a separate interview with Israel's Channel 12 news, Mr Andrés said "it was really a direct attack on clearly marked vehicles whose movements were known by everybody at the IDF [Israel Defense Forces]".
WCK released pictures of the victims...Source: World Central Kitchen
The bodies of six of the dead WCK workers have since been taken from Gaza into Egypt to be repatriated.
Their Palestinian colleague was buried in his hometown in Rafah, southern Gaza, on Tuesday.
Humanitarian aid to the Gaza Strip is in doubt after WCK - a key provider of aid to the territory - suspended operations.
The UN announced it was pausing movements at night for at least 48 hours to evaluate the security situation.
Israeli Prime Minister Benjamin Netanyahu has described the strike as unintentional.
"It happens in war, we check it to the end, we are in contact with the governments, and we will do everything so that this thing does not happen again," Mr Netanyahu said on Tuesday.
IDF Chief of General Staff Herzi Halevi called the incident a "grave mistake" and said "it shouldn't have happened," blaming the strike on misidentification.
US President Joe Biden has condemned the strike, accusing Israel of not doing enough to protect aid workers.
"The United States has repeatedly urged Israel to deconflict their military operations against Hamas with humanitarian operations, in order to avoid civilian casualties," Mr Biden said.
Three of the killed aid workers were British nationals. A Polish national, an Australian, a Palestinian and a dual US-Canadian citizen were also killed.
UK Prime Minister Rishi Sunak spoke to Mr Netanyahu on Tuesday. On the call, he described the situation in Gaza as "increasingly intolerable" and "demanded a thorough and transparent independent investigation" into the killing of the aid workers.
Mr Sunak added that Israel needed to end restrictions on humanitarian aid and protect civilians, according to a Downing Street statement.
In other reaction:
Australian Prime Minister Anthony Albanese said he had "expressed Australia's anger and concern" in a long phone call with Mr Netanyahu, and that he expected a "full and proper explanation for how this has occurred"
Polish Foreign Minister Radoslaw Sikorski said he had demanded an independent investigation from Israel Katz, his Israeli counterpart
Canadian Prime Minister Justin Trudeau said "full accountability" was needed, adding that it was "absolutely unacceptable for aid workers to be killed" by the IDF
Four days ago, WCK said that it had distributed 42 million meals in the Gaza Strip - dispatching more than 1,700 food trucks and also sending close to 435,000 meals by sea.
According to Cogat, the Israeli defence ministry body in charge of civilian policy in the Occupied Palestinian Territories, WCK is responsible for 60% of the non-governmental aid getting into the territory.
A second charity, the American Near East Refugee Aid (Anera), which was working closely with WCK, told the BBC it was also freezing its operations in Gaza.
More than 196 aid workers have been killed in Gaza since October, according to the US-funded Aid Worker Security Database, which records major incidents of violence against aid personnel. Not all have been killed in the line of duty.
Much of the Gaza Strip has been devastated during the Israeli military operations that began after Hamas gunmen attacked southern Israel on 7 October, killing about 1,200 people and seizing 253 hostages.
About 130 of the hostages remain in captivity, at least 34 of whom are presumed dead.
More than 32,916 people have been killed in Gaza since then, the Hamas-run health ministry says.
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