Scotland was held to a draw by Ukraine on Friday.
The game will be played at the Estadio de La Cartuja in Seville.
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Scotland will need their highest level to compete with Spain in Tuesday's Women's World Cup qualification, says head coach Pedro Losa.
Spain scored 35 goals and didn't concede a single one in Group B.
Scotland was two points behind the top seeds after Friday's disappointing draw at home to Ukraine.
"We will have to play our best match so far and be at our highest level in all aspects of our game," said Losa.
We will find opportunities to be competitive if we join with the spirit and character of the team and some little tactical details in our preparation. Taking pride in representing your country is the first responsibility.
Spain are ranked 10th in the world, 12 places above Scotland, and have been ruthless in the campaign, beating Hungary by seven and Ukraine by six, both on their travels.
The hosts have won 15 matches in a row with clean sheets.
There was extra motivation for him against his homeland, while he stressed that Spain and Scotland are in different moments of development.
He said that they're on a run of wins and performances at a good level. We don't feel there's a gap in that sense because we're going to try to compete in the one-off game. Spain needs to prove that their style can be used to achieve things.
Scots are ready for a daunting mission.
After the draw with Ukraine, veteran duos, including both of them from Manchester United, dropped out of the squad.
Last week, the team called up several players, including the one who scored the late goal in Glasgow, as well as Lana and Amy, while Tegan, Kelly Clark andLeah Eddie have also been drafted in.
Lee Alexander accepts that she may be busy in Seville but is urging her team-mates to get right in the faces of Spain.
She said that they didn't perform in the way they would have liked in the last few games.
We need to start putting that into proper performances because we have a good group of girls who have been together for a while. We've been getting points on the board but it's going to be difficult against Spain. We know how good they are.
"They can hurt us, but we can hurt them, so we need to go out and show what we can do, get right in their faces, and make sure we're competitive."
Scotland won the last time they met, at Euro 2017, but a two-goal margin was needed to progress from the group phase.
Scotland's last visit to Spain was a gut-wrenching Euro 2013 play-off defeat, with the decisive goal coming in the dying throes of extra-time.
