The 22-year-old Belgian ace will miss the Blues' second European final in as many years with a hamstring problem, leaving 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 they look 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 move from Lille last summer and was shortlisted for the PFA Player of the Year and Young Player of the Year awards, both won by Tottenham's Gareth Bale.
There had been hopes that Hazard would make the squad after travelling with the squad to Amsterdam but the youngster has been unable to recover in time from an injury he sustained 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 also remains doubtful.
Benitez added:" "John [Terry], we will see how he feels. We will decide tomorrow."