The 61-year-old coach is disappointed that the defender will be unable to take part in the European Championship and noted that the Barca striker is a serious doubt as well
The 34-year-old has been ruled out of this summer's showpiece as he will have to undergo surgery on a knee injury sustained in Barca's 4-0 victory over Espanyol on Saturday.
Puyol, who has 98 international caps to his name, was a key player in the Euro 2008 and World Cup 2010 winning squads, and Del Bosque admitted that the news is extremely disappointing.
"This is a big setback," he told AS.
The 61-year-old coach also cast doubt over David Villa's chances of making Spain's squad for the European Championship, as the Blaugrana striker is still recovering from a fractured tibia sustained at the Club World Cup in December.
"Villa is a serious doubt," Del Bosque noted. "We are worried, it is very hard to call up someone if they cannot play a single match."
Spain have been drawn with Italy, Republic of Ireland and Croatia at the tournament in Poland and Ukraine.