The January 18 crash in the southern region of Andalusia was already one of Europe's deadliest such disasters this century, casting doubt about the safety of train travel in Spain, which boasts the world's second-largest high-speed network after China.
A spokeswoman for the Andalusia regional government's health department told AFP that "one of the victims who was hospitalised has died" in the city of Cordoba's Reina Sofia hospital.
The department was not authorised to give any further details, she added.
"Andalusia remains heartbroken... what a tragedy, and what sorrow!," the region's leader Juanma Moreno posted on X.
The disaster struck when a train run by private firm Iryo derailed and smashed into an oncoming service operated by public company Renfe.
A preliminary report released last week suggested the track may have been cracked before the catastrophe.
On Thursday, King Felipe VI and Queen Letizia joined hundreds of mourners at a memorial Mass for the victims in the southern city of Huelva, the final destination of the train that suffered the most deaths.
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