The 22-year-old Belgian ace will miss the Blues' second European final in as many years with a hamstring problem, leaving Rafa Benitez without one of his key players.
"Hazard will not be available," Benitez told reporters.
The 15-goal attacker will be a key loss for the Blues as the London outfit looks to win back-to-back European trophies, following last year's Champions League victory.
Hazard has enjoyed a scintillating debut season for Chelsea following his 32 million pound move from Lille last summer and missed out on the PFA Player of the Year and Young Player of the Year awards to Tottenham's Gareth Bale.
There had been hopes that Hazard would make the squad after traveling with the squad to Amsterdam, but the youngster has been unable to recover in time from an injury he suffered in Chelsea's 2-1 win over Aston Villa on Saturday.
The Belgian ace is not the only selection issue for Benitez however as club skipper John Terry remains doubtful.
"John [Terry], we will see how he feels. We will decide tomorrow," Benitez said.