The Argentina forward, who was crowned FIFA World Player of the Year last month, missed Saturday's 1-1 La Liga draw at home to Villarreal after being granted an extra few days holiday over the winter break.
The La Liga leaders take on the 2007 Cup winners for the first of three clashes between the sides in 12 days. Sevilla, who have slipped to fifth in the standings, return to the Nou Camp after next week's second leg.
The Andalucians have lost their last two games and have been hit by a string of injuries to key players such as Brazil striker Luis Fabiano.
Like Barca, who have lost midfielders Seydou Keita and Yaya Toure to the African Nations Cup, Sevilla are also deprived of Mali striker Frederic Kanoute and Ivory Coast's Didier Zokora.
"They are a dangerous team whatever shape they are in," Barca reserve goalkeeper Jose Manuel Pinto told reporters.
"Toure and Keita are fundamental players for us, and I imagine the players they are losing (to the African Nations Cup) are important for them as well. It will be an evenly-balanced tie."
Giantkillers Alcorcon of the Segunda B (third tier), who dumped Real Madrid out in the last round 4-1 on aggregate, warmed up for Wednesday's visit of Racing Santander with a 2-1 league win over Real's youth team Castilla on Sunday.
Second division sides Recreativo Huelva and Celta Vigo host Atletico Madrid and Villarreal respectively on Wednesday, while La Liga heavyweights Valencia, who lifted the trophy in 2008, and Deportivo Coruna meet at the Mestalla.
Valencia climbed to third in the league after scrambling a last-minute winner for a 1-0 home win against Espanyol on Saturday, and could have midfielder David Silva back in the side after a month on the sidelines with a knee injury.
On Thursday, second division leaders Hercules are at home to Osasuna, second division Rayo Vallecano receive in-form Real Mallorca and struggling Malaga take on Getafe.
