Obama announced the additional funding Tuesday in a video aimed at the Syrian people.
"The relief we send doesn't say 'Made in America,' but make no mistake—our aid reflects the commitment of the American people," Obama said in the video, which was posted on the White House website.
The fresh funding brings the total U.S. humanitarian aid to Syria over two years to $365 million, according to the White House. Officials said the money was being used to immunize one million Syrian children, purchase winter supplies for a half million people, and to help alleviate food shortages.
The United Nations says more than 60,000 people have been killed since March 2011, when the clashes between the Syrian government and rebel forces started. The violence has also forced millions of people from their homes, destroyed the country's cities and created food and fuel shortages.
The U.S. has long called for Syrian President Bashar Assad to leave power and said the fall of his regime is inevitable. In addition to the humanitarian aid, the White House has also ratcheted up economic sanctions on Assad's regime and recognized the rebel-led Syrian Opposition Council as the legitimate representative of the Syrian people.
But mindful of the nation's war weariness, Obama is reluctant to directly intervene in the Syrian civil war.
"There are transitions and transformations taking place all around the world. We are not going to be able to control every aspect of every transition and transformation," Obama said Sunday in an interview with CBS' "60 Minutes." The president said his job is to protect the United States and engage where the U.S. can make a difference.
In his video released Tuesday, Obama said he was "under no illusions" about the situation in Syria, but continues to believe that Assad's regime will come to an end.
"The Syrian people will have their chance to forge their own future," Obama said. "And they will continue to find a partner in the United States of America."