Barcelona's defeat to Valencia raised early doubts about Quique Setien's appointment and gave Real Madrid the chance to move three points clear at the top.
Just a second slip up from Barcelona this La Liga season allowed Real Madrid and Atletico Madrid to close to within eight points of the leaders.
Jordi Alba will now have to rest and although he should be fit when Barcelona start their La Liga campaign he will miss the final of the European Supercup next Tuesday, which sees them take on their fellow Spaniards, Sevilla, who won last season's Europa League in Tiflis.
Jordi Alba has become an integral part of the Barca side since joining the club from Valencia in 2012.
Jordi Alba picked up a thigh injury against Ukraine and had to be replaced by Bayern Munich's Juan Bernat. The Barcelona player will miss Spain's friendly in Amsterdam against the Netherlands on Tuesday.
The Catalans were beaten for the first time in 21 games since Gerardo Martino took over in July when Ajax stunned them with a 2-1 win in the Champions League.
The injury will rule Jordi Alba out of Barcelona's Champions League opener at home against Ajax on Wednesday and its second group phase match at Celtic on Oct. 1.
The full-back limped off during Barca's victory over Sevilla with a thigh muscle tear.
The Spanish club says the players will be sidelined for 10 days while they recover after having procedures to treat the recurring problems of tonsillitis they both had had during recent weeks.
Barcelona have confirmed that Spain left-back Jordi Alba will undergo the same procedure in Spain, also Friday.
The operations had been scheduled to take place after the Confederations Cup final in Rio de Janeiro on Sunday, when Brazil crushed Spain 3-0 with the help of a sizzling left-footed goal by Neymar.
Nigeria were ultimately left to rue last Thursday's damaging 2-1 defeat by Uruguay, having begun their Confederations Cup campaign with a breezy 6-1 thrashing of Tahiti. It was the teams' first meeting since Nigeria beat Spain 3-2 in the group phase of the 1998 World Cup, and if the world champions were out for revenge, they were quick to draw ...
The nation-wide mass demonstrations, which have sometimes turned violent, are taking place as Brazil is hosting teams from around the world for the Confederations Cup, a dry run for next year's World Cup being held in six host cities.
The Spaniards dominated throughout but after Pedro Rodriguez's opener deflected had cause to be grateful to Pedro Soldado's 32nd-minute clincher as Luis Suarez netted a late freekick for the Uruguayans.
Alonso has struggled with injuries in the second-half of the season and has not been given the all-clear by Real Madrid's doctors to take part in the competition which starts on June 15.
Busquets has also been ruled out to a pelvic injury, whilst Iniesta and Alba's absences are purely down to the decision of the management staff.
Xavi Hernandez and Jordi Alba will travel with Barcelona for their Champions League quarter-final first-leg at Paris Saint-Germain on Tuesday after returning to full training on Sunday.
Vicente Del Bosque's side have not lost a competitive match since the 2010 World Cup in South Africa, but two consecutive home draws have seen them fall behind France in this qualifying campaign.