Glencore in July this year had contacted Rio Tinto regarding a potential merger of both the mining giants, which was rejected in August.
"The board of Rio Tinto notes the recent press speculation regarding a possible combination of Rio Tinto and Glencore. The Rio Tinto board confirms that no discussions are taking place with Glencore," Rio Tinto said in a statement.
Media reports had speculated a possible merger of both the companies to create a USD 160 billion mining and trading giant.
The board's rejection was communicated to Glencore in early August and there has been no further contact between the companies on this matter, it said.
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"Rio Tinto remains focused on the successful execution of its strategy, which the board of Rio Tinto is confident will continue to deliver significant and sustainable value for shareholders," it added.
Rio Tinto chairman Jan du Plessis said under the leadership of Sam Walsh and Chris Lynch, Rio Tinto has made significant progress in refocusing and strengthening its business.