The newly hardened view of Moscow comes as the president has tried to shake suspicions about the motives behind his campaign calls for warmer relations.
Tillerson met President Vladimir Putin and Foreign Minister Sergei Lavrov for discussions that were meant to tackle, primarily, the fight against Daesh and safe zones, but that were consumed by the chemical attack in Syria.
US President Donald Trump described the relationship between the two countries as at an "all-time low".
World leaders had wanted an investigation into the chemical attack in Idlib last week, which prompted a retaliatory USA missile strike.
The agenda for the meeting wasn't announced.
Tillerson again said the USA has "conclusive" evidence Syrian President Bashar al-Assad's regime planned and carried out a chemical attack on its civilians last week.
Russian Federation - refusing to concede that Assad - was behind the attacks.
The meeting was steeped in geopolitical intrigue, with neither country saying ahead of time if it would take place or not.
Earlier Wednesday, Russia's top diplomat accused the United States of carrying out an unlawful attack against Syrian President Bashar Assad's forces. Russian Federation says rebels were responsible for whatever chemical agent was used, which the Trump administration calls a disinformation campaign. "How do you know that?"
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"We have information that a similar provocation is being prepared... in other parts of Syria including in the southern Damascus suburbs where they are planning to again plant some substance and accuse the Syrian authorities of using chemical weapons", he said on Tuesday. Tillerson conceded the two world powers had "sharp differences" that have obstructed cooperation but voiced optimism that their talks could narrow those differences. "Also, we had held talks in the Kremlin before that", Lavrov said.
Until the chemical attack, the Trump administration had sought to step back from the U.S. position that Assad should leave power. "Even if we have them, we wouldn't use them", he said.
PRESIDENT DONALD TRUMP: Right now, we're not getting along with Russian Federation at all.
REX TILLERSON: We do think it's important that Assad's departure is done in an orderly way, an orderly way, so that certain interests and constituencies that he represents feel that they have been represented at the negotiating table for a political solution.
But Lavrov, on Thursday, offered a speck of optimism for the future of the relationship between the two Cold War foes.
"It is encouraging to some extent that Rex Tillerson confirmed yesterday that [the United States still holds] the opinion that there is no alternative to the political process [of the resolution of the Syrian crisis] despite all the recent negative developments", he said. It lasted for about two hours.
Trump said it was "certainly possible" that Putin knew about the attack, blamed on Assad, indicating Russian officials were present at the source airbase, which Trump later bombed.
The United States is "not getting along with Russian Federation at all", Trump said at a press conference at the White House. Interestingly enough, in the same press conference where Trump said relations were poor, NATO Secretary General Jens Stoltenberg addressed the comparison.
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