Foreign Minister Jose Manuel Garcia-Margallo said his country had racked up "big losses" from the sanctions, after a meeting with his Russian counterpart Sergei Lavrov in Moscow.
"No one is benefiting from these sanctions which are causing significant prejudice to the Spanish economy," he said during a joint news conference.
Last year the EU and United States slapped economic sanctions on Moscow which they accuse of arming and backing the rebels - a charge denied by the Kremlin.
Moscow hit back with a retaliatory ban on food products from countries which imposed sanctions such as Spain which was a major supplier of fresh fruit and vegetables to Russia.
Garcia-Margallo in February said the EU as a whole had lost €21 billion ($22.6 billion) in lost exports as a result of the sanctions.
Spain, a prime destination for Russian tourists before the collapse of the ruble to nearly half its value against the dollar, has also been "strongly affected by a fall" in visitors from Russia.
Nevertheless the Spanish economy expanded by 1.4 percent in 2014, after five years of recession or no growth.
Garcia-Margallo said he did not expect to see an increase in sanctions against Moscow due to the "good news" coming from Ukraine, where both rebels and government troops have reportedly withdrawn most heavy weapons following a ceasefire agreed in Minsk in February.
Lavrov praised Spain for its "measured position in this difficult period for Russia and the EU" whose relations have plunged to their lowest levels since the Cold War over the Ukraine crisis.
"The normalisation of our cooperation is in Russia's interests," said Lavrov, adding that he was convinced "good sense would prevail" in the EU.