Luis de la Fuente's side will enter the contest off the back of a 1-0 success over Denmark, while Serbia recorded an eye-catching 2-0 victory over Switzerland last time out to move onto four points in the section.
Match preview
Spain were far from their best against Denmark on Saturday evening, and it did appear that La Roja might have to settle for a goalless draw, but Martin Zubimendi came up with the game's only goal in the 79th minute.
The Euro 2024 winners therefore moved to the top of the section ahead of Denmark, boasting seven points from their opening three matches, and they will now be bidding to move onto 10 points ahead of their final two games in the group stage, which will come against Denmark and Switzerland next month.
Spain are the reigning Nations League champions, having triumphed in the 2022-23 competition, beating Croatia in the final to secure the trophy for the first time.
La Roja are unbeaten since a 1-0 loss to Colombia in March, but they were held to a goalless draw by Serbia in the reverse match last month, and the visitors will enter this clash off the back of an excellent result.
Spain are also missing some vital players through injury, meanwhile, and are sweating over the fitness of Lamine Yamal for this contest due to the issue that the Barcelona wonderkid picked up against Denmark.
Serbia failed to make it out of the group stage of Euro 2024, picking up just two points from their three matches, but they have shown some positive signs during their 2024-25 Nations League campaign.
Dragan Stojkovic's side opened their Group 4 challenge with a goalless draw against Spain, but they were unable to build on that result, suffering a 2-0 defeat to Denmark in their second game in September.
The Eagles bounced back in impressive fashion on Saturday, though, beating Switzerland 2-0, with Aleksandar Mitrovic adding to an own goal from Nico Elvedi to secure all three points.
Serbia are third in their group, two points behind second-placed Denmark and three from leaders Spain, so a victory on Tuesday night would put them in an interesting position ahead of their final two group games in November.
Stojkovic's side have only lost one of their last five games in all competitions, but their first-ever meeting with Spain in May 2012 ended in a 2-0 defeat, with Adrian Lopez and Santi Cazorla on the scoresheet.
Spain Nations League form:
D
W
W
Spain form (all competitions):
W
W
W
D
W
W
Serbia Nations League form:
D
L
W
Serbia form (all competitions):
L
D
D
D
L
W
Team News
Spain are currently missing a number of important players through injury, with Unai Simon, Dani Carvajal, Robin Le Normand, Rodri, Dani Olmo, Gavi and Nico Williams among those currently unavailable for selection.
La Roja, as mentioned, will also need to assess Yamal, and it would not be a surprise to see the Barcelona teenager withdraw from the squad due to the minor issue that he suffered against Denmark.
Alex Baena would be the favourite to come in for Yamal, but it might otherwise be the same side that took to the field against Denmark, with Alvaro Morata, who has 36 goals for his country, leading the line.
As for Serbia, head coach Stojkovic could decide to select the same XI that started the impressive win over Switzerland, with Mitrovic continuing in the final third of the field.
Mitrovic has scored 59 times in 95 caps for his country, while the 30-year-old has been in excellent form for Al Hilal this season, managing 14 goals in 11 appearances in all competitions.
Nikola Milenkovic and Sasa Lukic should also keep their spots in the side, while Porto's Marko Grujic is expected to feature in the XI despite being withdrawn at the interval of the clash with Switzerland.
Spain possible starting lineup:
Raya; Porro, Vivian, Laporte, Cucurella; Zubimendi, Ruiz; Baena, Pedri, Oyarzabal; Morata
Serbia possible starting lineup:
Rajkovic; Erakovic, Milenkovic, Pavlovic; Nedeljkovic, Maksimovic, Grujic, Biramcevic; Samardzic, Mitrovic, Lukic
We say: Spain 2-1 Serbia
This has the potential to be a very tricky match for Spain, as Serbia can be a tough nut to crack, and the hosts will be missing a number of important players. It would not be a surprise to see a draw here, but we just have a feeling that De la Fuente's side will be able to navigate their way to a huge three points.