The measures, asset freezes and travel bans, are part of a package of sanctions, including an arms embargo which went into effect on Tuesday, as part of EU efforts to try to force Syria to end violence against anti-government protesters.
EU governments decided not to target the president for now, in what diplomats said was a bid to introduce punitive measures gradually.
But Bashar al-Assad, grappling with the most serious challenge to his 11-year rule, could face EU sanctions soon, they said.
Failure to put Assad on the list underlines splits in the EU over the effectiveness of an embargo in restraining his government's actions.
Live Blog Syria
Sources said Germany and Spain opposed adding the president, over-riding strong backing for such a move from France and others.
Included was Rami Makhlouf, a cousin of Assad, who owns Syria's largest mobile phone company, Syriatel, and several large firms in the construction and oil sectors.
The EU said in its official journal that he "bankrolls (Assad's) regime, allowing violence against demonstrators".
In 2008, the United States imposed sanctions against him because of corruption allegations.
Also affected is Ali Mamlouk, head of the General Intelligence Service, and Adulfattah Qudsiyeh, who runs military intelligence.
Call for more protests
Syrian activists called for countrywide protests on Tuesday in solidarity with thousands of anti-regime activists rounded up by the security forces, setting the scene for another round of bloody clashes.
"Demonstrations will continue every day," said the Syrian Revolution 2011 Facebook page, which has been a motor of the protests.
It called for "a Tuesday of solidarity with prisoners of conscience held in the jails of the criminal Syrian regime".
Street demonstrations are persistently dispersed with violence by the security forces, who also make mass arrests, according to rights activists, who say more than 600 people have been killed and 8,000 jailed or gone missing in the eight-week crackdown.
In the latest security force bid to crush the anti-regime protest movement, troops went house to house in the coastal city of Baniyas on Monday, rounding up thousands of men, activists said.
Rami Abdul Rahman, head of the London-based Syrian Observatory for Human Rights, said water, electricity and telephone lines were cut off in Baniyas, on Syria's northwest Mediterranean coast.
"Thousands of men, including youths, have been rounded up by the army and security forces... to be interrogated and they are being beaten. More than 400 are still being held," he said.
A Syrian senior government official said in an interview with the New York Times, meanwhile, that she believes Assad's embattled government had ridden out the worst of the uprising.
"I hope we are witnessing the end of the story," Bouthaina Shaaban, an adviser to President Assad who often serves as a spokeswoman, told the US paper in an hour long interview.
"I think now we've passed the most dangerous moment. I hope so, I think so," Shaaban said, giving a glimpse at the mindset of a 40-year-old regime that has barred most foreign journalists from Syria since the beginning of the uprising.
"We want to use what happened to Syria as an opportunity," Shaaban added. "We see it as an opportunity to try to move forward on many levels, especially the political level."
The Times reporter was allowed in the country for a few hours, the report added.
Syria's upheaval began on March 18 when protesters, inspired by revolts across the Arab world, marched in the southern city of Deraa.
Assad initially responded with vague promises of reform, and last month lifted a 48-year-old state of emergency. But when the demonstrations persisted he sent the army to crush dissent.
Meanwhile, concerns remain for the welfare of Dorothy Parvaz, an Al Jazeera journalist, who has not been heard from since she arrived in the capital, Damascus, on April 29.
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