Tuesday's announcement falls under Presidential Drawdown authority and marks the 40th such commitment by the United States since August of 2021.
Military includes weapons, ammunition, vehicles and other equipment required by Ukraine in its efforts to repel an invasion by Russia.
The latest aid package also includes surface-to-air missile systems and anti-aircraft systems.
"It includes key capabilities to aid Ukraine's efforts to retake its sovereign territory and support Ukraine's air defenders as they bravely protect Ukraine's soldiers, civilians, and critical infrastructure, as well as artillery, anti-armor systems, and ammunition valued at up to $325 million," the Defense Department said in a statement.
"The United States will continue to work with its allies and partners to provide Ukraine with capabilities to meet its immediate battlefield needs and longer-term security assistance requirements."
In November, U.S. officials gave an additional $400 million of military aid to Ukraine under presidential drawdown authority.
Last week, the Pentagon pledged an additional $2.1 billion worth of military aid for Ukraine.
That assistance fell under the Defense Department's Ukraine Security Assistance Initiative, and does not deplete American weapons stockpiles in the same way as a presidential drawdown of equipment.
The United States has pledged more than $40.4 billion under that initiative since the beginning of President Joe Biden's administration.
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