The new list - published on the Official Journal of the European Communities (OJEC) at 8.30 a.m. GMT - places "restrictive measures" on 28 people or organizations, including Russia's First Deputy Minister of Defense, Arkady Bakhin.
Also on the list was Deputy Minister of Defense Anatoly Antonov and Andrei Kartapolov, the deputy chief of the general staff of the Russian armed forces. The sanctions are due to come into effect immediately.
The new penalties are part of an ongoing program by the European Union, but come just days after a cease-fire was announced in Ukraine. Military conflict with Russian separatists has been one of the biggest factors weighing on markets in recent months, but despite the cease-fire some of the rebels have not observed the truce , according to reports.
Ukraine was thrown into turmoil at the start of last year, after protests between anti-government and pro-EU demonstrators led to a change of leadership.
Tensions on the streets of Kiev turned into military conflicts on the eastern border, with Moscow accused of aiding pro-Kremlin rebels in the region. Moscow continues to deny the involvement of Russian troops in the conflict.
Despite these denials, the tensions have hit Russia's economy hard. It is expected to fall into a recession in the coming year on the back of international economic sanctions from both the U.S. and Europe, combined with a dramatic fall in oil prices.
The Russian ruble fell sharply against the dollar after the news of more sanctions Monday, despite appreciating much of the morning session.
The economic sanctions now mean Western asset freezes and travel bans for yet more Ukrainians and Russians.
As well as commanders of armed separatist group in the region, the list also includes Iosif Kobzon, a Russian singer, who the EU has accused of making statements supporting separatists and voting in favor of the annexation of Crimea.
The new sanctions list was drawn up on February 9, according to the EU, before the cease-fire announced Friday.
Timothy Ash, head of emerging markets at Standard Bank, said in a note that the sanctions process was often slow and doesn't usually "dovetail" with the political situation.
While Kit Juckes, global head of foreign exchange strategy at Societe Generale, told CNBC Monday that it wasn't surprising pressure on Russia was being maintained in economic terms.
"(It has to) make sure that everybody understands that we have to get beyond just a cease-fire, we have to get through to an end to the fighting and find some lasting solution," he said.
"We should expect the sanction structure to remain in place until that's the case. If you back down from that now...the belief would be that everything would fall apart."