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Arsenal's Champions League fate will be decided in their final Group C match after group winners Barcelona earned a comfortable win at Emirates Stadium.
But the Gunners, in second place, three points above Hoffenheim, still have their future in their own hands before next week's meeting with the Germans.
Aitana Bonmati, Jennifer Hermoso and Fridolina Rolfo put defending champions Barcelona in control at half-time.
Hermoso then added her second to make it 4-0 with 15 minutes remaining.
Arsenal needed to avoid defeat to secure their progress to the last eight with a game to spare in Group C - or hope Hoffenheim failed to beat HB Koge.
Hoffenheim won 2-1 to keep their hopes alive, but Jonas Eidevall's Arsenal side will be confident of sealing progress against opponents they beat 4-0 at home on matchday two.
Barcelona dominated possession and territory but Jordan Nobbs shot narrowly over after regaining possession in midfield with Arsenal's first attempt with 10 minutes played.
Manuela Zinsberger produced an excellent save to deny Bonmati, but the Spaniard rolled in the visitors' first after 22 minutes following a mistake by Jennifer Beattie.
Ballon d'Or runner-up Hermoso added a second from Rolfo's low cross seven minutes later and, while some resolute Arsenal defending prevented further damage, Rolfo's stunning finish from outside the penalty area in first-half added time effectively sealed victory.
Unbeaten Super League leaders Arsenal suffered only their second loss of the season in Sunday's delayed 2020-21 FA Cup final against Chelsea - their other arriving at the hands of a Barcelona side considered "the world's best" by Arsenal boss Eidevall.
Eidevall said he did not recognise his own team in the 3-0 defeat at the weekend, which ended an eight-match unbeaten run, although he will have been pleased with the second-half response from his players on Thursday.
Marta Torrejon hit the crossbar and Zinsberger again came to the rescue as a relentless Barcelona were initially frustrated in their search for a fourth goal - before Hermoso's scrappy second crept over the line.
Barcelona had already secured their quarter-final spot but confirmed their place as group winners with a dominant display, in which they had 20 shots to Arsenal's six.
The Gunners now host winless Leicester City in the Super League on Sunday before their crucial final group match at Hoffenheim on Wednesday.
Line-ups
Arsenal Women
Formation 4-3-3
- 1Zinsberger
- 16Maritz
- 5BeattieSubstituted forBoye Sørensenat 80'minutes
- 7Catley
- 15McCabe
- 10Little
- 12MaanumSubstituted forWältiat 60'minutesBooked at 60mins
- 8NobbsSubstituted forIwabuchiat 60'minutes
- 14Parris
- 19FoordBooked at 26minsSubstituted forMiedemaat 60'minutes
- 9MeadBooked at 90mins
Substitutes
- 3Wubben-Moy
- 4Patten
- 11Miedema
- 13Wälti
- 18Williams
- 20Boye Sørensen
- 22Schnaderbeck
- 23Iwabuchi
- 29Goldie
Barcelona Femenino
Formation 4-3-3
- 1Paños
- 8TorrejónBooked at 20minsSubstituted forFernándezat 64'minutes
- 2Paredes
- 17Pereira
- 16RolföSubstituted forSerranoat 85'minutes
- 14Bonmatí
- 11PutellasBooked at 30mins
- 9Caldentey OliverSubstituted forOuahabiat 80'minutes
- 18CrnogorcevicSubstituted forEngenat 65'minutes
- 10Hermoso Fuentes
- 22MartensSubstituted forHansenat 65'minutes
Substitutes
- 3Fernández
- 5Serrano
- 6Pina
- 7Hansen
- 13Coll
- 15Ouahabi
- 23Engen
- 24Font
- 29Pérez
- 32Vignola
Referee:Anastasia Pustovoytova
Live Text
Match ends, Arsenal Women 0, Barcelona Femenino 4.
Second Half ends, Arsenal Women 0, Barcelona Femenino 4.
Offside, Arsenal Women. Stephanie Catley tries a through ball, but Nikita Parris is caught offside.
Corner, Arsenal Women. Conceded by Irene Paredes.
Attempt blocked. Katie McCabe (Arsenal Women) left footed shot from the left side of the box is blocked. Assisted by Beth Mead.
Beth Mead (Arsenal Women) is shown the yellow card.
Foul by Irene Paredes (Barcelona Femenino).
Beth Mead (Arsenal Women) wins a free kick in the defensive half.
Corner, Barcelona Femenino. Conceded by Stephanie Catley.
Corner, Barcelona Femenino. Conceded by Stephanie Catley.
Attempt missed. Aitana Bonmatí (Barcelona Femenino) right footed shot from a difficult angle on the right is close, but misses to the right. Assisted by Alexia Putellas.
Substitution, Barcelona Femenino. Melanie Serrano replaces Fridolina Rolfö.
Attempt missed. Mana Iwabuchi (Arsenal Women) left footed shot from the left side of the box misses to the left following a fast break.
Attempt saved. Beth Mead (Arsenal Women) right footed shot from the centre of the box is saved in the bottom left corner. Assisted by Lia Wälti.
Irene Paredes (Barcelona Femenino) wins a free kick in the defensive half.
Foul by Katie McCabe (Arsenal Women).
Substitution, Arsenal Women. Simone Boye Sørensen replaces Jennifer Beattie.
Substitution, Barcelona Femenino. Leila Ouahabi replaces Mariona.
Foul by Jana Fernández (Barcelona Femenino).
Beth Mead (Arsenal Women) wins a free kick in the defensive half.