Southampton suffered defeat in their Audi quattro Cup semi-final, as Valencia progressed courtesy of Pablo Piatti’s solitary goal.
Saints twice went close through Sadio Mané and José Fonte as they both hit the crossbar, but Piatti’s goal moments after was enough to seal the Spanish side’s place in the final.
Cuco Martina was handed his first start in a Southampton shirt as he lined up at left-back, with Mané also starting on his return to the Red Bull Arena.
Harrison Reed, Jack Stephens, Lloyd Isgrove and Juanmi were the only four players to have featured in the opening pre-season game against RB Leipzig on Wednesday.
Fonte twice did well to stop Valencia from working a scoring opportunity in the opening stages, firstly cutting out Alvaro Negredo’s cut-back and then clearing away Antonio Barragan’s cross as Negredo lurked.
Mané, who was receiving a spicy reception on his return to Austria, was the first to test the Valencia defence. The 23 year-old drove to the byline and looked to find Graziano Pellè, but his cross was just out of the Italian’s reach.
Soon after, Saints were within a whisker of going ahead on two occasions. After Mané had headed against the crossbar, Fonté followed suit from the rebound.
The two close shaves seemed to spark the Spanish opposition into life, and they went ahead with 23 minutes on the clock. Barragan’s cross caused havoc in the box and despite an excellent save from Paulo Gazzaniga, Piatti fired home the rebound.
Gazzaniga had more fortune just after the half-hour mark when he denied Dani Parejo and Piatti in quick succession as the Spaniards looked to double their lead.
At the other end, Isgrove tested Jaume from distance, but saw his effort held well by the Valencia stopper at his near post in what proved to be Saints' last chance of the game.
The defeat meant Saints would face Red Bull Salzburg, the tournament hosts, in the third versus fourth play-off, while Valencia would take on Werder Bremen in the final after their 4-0 semi-final victory.
Philipp Bargfrede handed the German side the lead in the other semi-final when he coolly slotted home from just inside the penalty area, before Anthony Ujah doubled their lead with a clever flick.
Ulisses Garcia and Franco Di Santo added further goals for Bremen as they progressed to the final, consigning Red Bull to the third versus fourth play-off with Saints.
Southampton team: Paulo Gazzaniga, Jason McCarthy (Sam Gallagher 42), José Fonte (c), Jack Stephens, Cuco Martina, Harrison Reed, Victor Wanyama, Lloyd Isgrove, Juanmi, Sadio Mané, Graziano Pellè.
Southampton substitutes: Kelvin Davis (GK), Armani Little, Jordan Turnbull, Sam McQueen, Jake Hesketh, Will Wood.
Valencia team: Jaume, Antonio Barragán, Rúben Vezo, Lucas Orban, José Luis Gayà, Dani Parejo (c), Javi Fuego, Enzo Pérez, Pablo Piatti, Rodrigo, Alvaro Negredo.
Valencia substitutes: Ferran Ferri (GK), Sergio Ayala, Ibrahim Diallo, Zakaria Bakkali, Rodrigo de Paul, Alex de Groot, Santiago Mina
Goals: Pablo Piatti (23)