Biden spoked to the Ukrainian leader over telephone and "made clear that the US is prepared to sanction those officials responsible for the violence", the White House said in a statement.
He called upon President Yanukovych to immediately pull back all security forces police, snipers, military and paramilitary units, and irregular forces, it said.
"The Vice President urged President Yanukovych to take immediate and tangible steps to work with the opposition on a path forward that addresses the legitimate aspirations of the Ukrainian people."
Biden underscored that the US supports an independent, democratic Ukraine that pursues the future its people choose. The US leader also called the Polish Prime Minister to consult on the situation in Ukraine.
They discussed steps the United States, Poland, and the European Union are taking to support an end to the violence and a political solution that is in the best interests of the Ukrainian people, the statement said.
In a late night statement, Secretary of State John Kerry asked the Ukrainian Government to stop violence.
"The violence must stop. We unequivocally condemn the use of force against civilians by security forces, and urge that those forces be withdrawn immediately," Kerry said as he expressed his anger and anguish having watched renewed violence on the streets of Kiev.
"The people of Ukraine deserve far better than senseless death and suffering on the streets of one of Europe's great cities," he said adding that the people of Ukraine and the international community will hold to account those who are responsible for what has occurred.
The US has already begun implementing sanctions through travel bans on Ukrainians responsible for the violence, he said.
Noting that protesters should exercise their rights peacefully, he urged the Ukrainian military to remain true to its non-political heritage and its professional traditions.
"There is no time for brinksmanship or gamesmanship. President Yanukovich must undertake serious negotiations with opposition leaders immediately to establish a new interim government that will have broad support," he said.