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In the video, Ayman al-Zawahiri referenced events that took place after the rumours of his alleged death started to circulate in December, giving some pretty strong evidence that he's still alive.
The 70-year-old Egyptian previously operated under Osama Bin Laden, but rose higher up the group after Bin Laden was killed.
His whereabouts are currently unknown, but he has been thought to be hidden somewhere within tribal Pakistan.
Al-Zawahiri is one of the most wanted men in the world, with the US Department of State's Rewards for Justice Program offering a $25 million reward for information as to his whereabouts.
The 60 minute video was reference in a tweet by a US-based intelligence group called SITE, which tracks the online presence of white supremacist and jihadist groups.
SITE director Rita Katz wrote: "Amid rumours of his death, Al-Qaeda leader Ayman al-Zawahiri shown in a new 60-minute video, this time offering some evidence that he is not dead - particularly, reference to events after December when rumours of death surfaced."
This particular video was called 'Jerusalem will not be Judaized', and was seemingly put out by the propaganda wing of Al Qaeda, As Sahab Media.
In the video, the high-ranking member talked about a raid on a Russian military base in Syria performed by the Al Qaeda aligned Hurras al-Deen group, which they said took place on January 1.
Of course, if the rumours of his death - which to swirl in November - were true, then he couldn't possibly have known about that.
Zawahiri is believed to have taken over Al Qaeda after the death of Osama Bin Laden at the hands of US Navy Seals.
The US military special forces unit raided a compound in Abbottabad, Pakistan, and killed Bin Laden on May 2 2011.
However, Katz when on to say that whilst al-Zawahiri did discuss the US withdrawal from Afghanistan, he did not mention the Taliban's return to power.
She continued: "However, Zawahiri doesn't mention Taliban's Afghanistan victory, and his talk of US 'making its exit from Afghanistan' could have been said early as Feb 2020 upon Doha Agreement,
"Thus, he could still be dead, though if so, would have been at some point in or after Jan 2021."
Still, she added that it's not possible to say definitively whether he is alive or dead, because 'intelligence agencies have, as of yet, offered no proof or solid assessments that Zawahiri is dead, leaving the question of his current status in the air.'
"Tragic as it is to say, this 9/11 anniversary is a uniquely positive one for al-Qaeda. As one AQ supporter wrote, the US leaving Afghanistan is validation of Bin Laden's vision and the 'blessed' 9/11 attacks, and that 'Afghanistan is the beginning'," she said.
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