Arbeloa was expected to start after a fine season with Madrid but had to pull out of the world and European champions' first training session at their base near Gdansk with a back problem.
Should the former Liverpool man not recover in time to face the Italians on Sunday, Atletico Madrid's Juanfran could make only his second international appearance on the right side of the defence after performing strongly on his debut in the friendly against Serbia last week.
On a brighter note for Del Bosque, Barcelona midfielder Cesc Fabregas has recovered from the hamstring injury which kept him out of Sunday's 1-0 win over China and will be fit for selection on Sunday barring any further setbacks.
Italy's build-up has been blighted by more allegations of match-fixing but Spain's Javi Martinez does not believe the distraction will have a huge bearing once the tournament begins.
He told Radio Marca: "It will be a difficult match. It is true that right now they are going through delicate moments with the match-fixing topic but it is a nation that always plays with pride."
The Athletic Bilbao midfielder was part of the squad which won the World Cup in 2010 and the Under-21 European Championship last summer and he is looking forward to the challenge of winning three major tournaments back to back.
"I feel fortunate to be able to play with this team. I debuted two years ago and now we have the opportunity to make history in this European Championship. This motivates us a lot. I am prepared to play whenever I am called."