Reacting for the first time after the last week's attack, Rohey Hydara said: "I am saddened and shocked by what Khalid has done. I totally condemn his actions."
Hydara released the statement through Scotland Yard, a day after Masood's mother had similarly condemned the attack which claimed four lives and injured 50 others.
The 39-year-old expressed condolences to the families of the victims that have died, and wished a speedy recovery to all the injured.
"I would like to request privacy for our family, especially the children, at this difficult time," she said.
"I wish to make it absolutely clear, so there can be no doubt, I do not condone his actions nor support the beliefs he held that led to him committing this atrocity," she had said, in reference to her son born Adrian Russell Ajao before he converted to Islam.
The Metropolitan Police has said Masood had had a clear interest in jihad and his methods echoed the rhetoric ofthe Islamic State but there was no evidence he acted as part of the terrorist group.
"There is no evidence that Masood was radicalised in prison in 2003, as has been suggested; this is pure speculation at this time. While I have found no evidence of an association with IS or AQ (Al Qaeda), there is clearly an interest in Jihad."
"I know when, where and how Masood committed his atrocities, but now I need to know why. Most importantly, so do the victims and families," he said.
The evidence into the deaths of the victims and the attacker has been presented by the Met at Westminster Coroner's Court yesterday.
"At this stage the Met will not confirm provisional cause of death," the Met Police said.
Masood's inquest will open and adjourn at the same location on Thursday.
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