Quarter-finalists last year, the home side finished second to Real Madrid during the autumn group stage, having been beaten by the Spanish giants both home and away.
Three wins from their other four matches saw them through, though, and Napoli now face opponents who are back in the knockout phase for the first time in three seasons - Barcelona finished first in Group H, just ahead of Porto.
Here, Sports Mole takes an in-depth look at the head-to-head record and previous meetings between the two sides.
Head-to-head record
Previous meetings: 6
Napoli wins: 0
Draws: 2
Barcelona wins: 4
Regularly pitched together in European competition over the past few years, Napoli and Barcelona have both found the net on each occasion, with the Italian side yet to get the better of their Spanish counterparts.
After holding the Catalan club to a 1-1 draw on their first competitive visit to Camp Nou in February 2022, the Partenopei were well beaten back in Campania one week later, when hopes of progressing to the Europa League's latter stages quickly crumbled.
The visitors wasted no time in taking the lead through Jordi Alba, before Frenkie de Jong curled home Barca's second from distance. Though Napoli halved their deficit soon after - following Marc-Andre ter Stegen's foul on Victor Osimhen, and Lorenzo Insigne's penalty conversion - Xavi's side struck again before half-time: Gerard Pique restored Barcelona's two-goal advantage.
Within 15 minutes of the re-start, Pierre-Emerick Aubameyang found a fourth, rendering Matteo Politano's late strike a mere consolation for the disappointed fans at Stadio Maradona.
Two years earlier, the clubs had met in the Champions League's last 16, in a tie which was played either side of a six-month hiatus due to COVID-19 travel restrictions.
In February 2020, Dries Mertens scored for Napoli, and Antoine Griezmann replied for Barcelona in Naples, but a pandemic intervened before the two-legged contest could reach its conclusion.
At an almost empty Camp Nou, nearly half a year later, Barca set up a quarter-final clash against Bayern Munich thanks to a frantic first half that saw Clement Lenglet, Lionel Messi and Luis Suarez all find the net.
Messi was denied a second for handball, having already produced a superb run and sublime finish to put his team 2-0 up on the night; the Argentinian wizard also won the penalty that Suarez coolly converted.
There was still time for Insigne to respond from the spot during first-half stoppage time, but the second half was far less eventful and Barcelona ultimately ran out 4-2 winners on aggregate.
Previous meetings
Feb 24, 2022: Napoli 2-4 Barcelona (Europa League Qualifying Playoffs)
Feb 17, 2022: Barcelona 1-1 Napoli (Europa League Qualifying Playoffs)
Aug 08, 2020: Barcelona 3-1 Napoli (Champions League Round of 16)
Feb 25, 2020: Napoli 1-1 Barcelona (Champions League Round of 16)
Aug 10, 2019: Barcelona 4-0 Napoli (friendly)
Aug 07, 2019: Napoli 1-2 Barcelona (friendly)
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