The 66-year-old suggested the center back will probably not be in the right shape for the tournament in Brazil, but talked up Thiago Alcantara's chances
Vicente del Bosque's side romped to victory four years ago with the Blaugrana veteran a rock in the back four, but he will be 36 when this summer's tournament in Brazil gets under way.
With question marks over his fitness and only one appearance in La Liga in 2014, Grande thinks the time may have come for La Roja to part ways with the defender who has 100 international caps to his name.
"Puyol has always been a very important player for us, not only on the pitch but also in pre-match meetings," Grande told La Xarxa. "He has experience, style, guts, behavior and enthusiasm.
"I haven't ruled him out completely yet, but obviously he needs to play a lot more and be more involved in Barca's key matches.
"He hasn't been completely ruled out, as there's still three months until the World Cup, but it is looking very difficult for him."
Del Bosque's right-hand man, who denied that La Roja's team is an aging one, also revealed that Thiago Alcantara's performances for Bayern Munich this season are likely to see him secure a place in Spain's squad for the World Cup.
"We're not a veteran team," Grande added. "With the exception of a couple of players, the rest are at a great age.
"In principle, the squad that went to two European Championships and the World Cup will go to Brazil. That is the plan as long as there are no major injuries. There might be a couple of changes, but it will be 80% to 90% the same team that has given us so much joy in the past.
"Out of the youngsters, Thiago is the one with the most experience. He plays frequently at Bayern, he's a regular in all their games.
"Out of all the young players we have on our radar, he has the best chance of going to the World Cup."