Hamburg have been fighting to return to the top-flight football after they were relegated for the first time in 2018 and will now spend a third successive season in the second tier after suffering a 5-1 home defeat to Sandhausen on Sunday night.
The dramatic nature of the loss stems from the fact that Hamburg needed just a point to qualify for the promotion play-off spot which was taken by Haddenheim, who finished in third spot despite suffering a 3-0 defeat to Arminia Belefeld.
The ex-German champions were the last team to have played every season of the Bundesliga since the league was founded in 1963, until "der Dino" as it's affectionately known were finally relegated after a number of close calls.
Haddenheim will now play Hamburg's bitter rivals Werder Bremen in a two-leg promotion/relegation play-off.
Hamburg found themselves 2-0 down after 21 minutes through an own goal from Rick van Drongelen and Kevin Behrens's goal before an Aaron Hunt penalty gave them hope.
However, a Behrens added the third from the penalty spot before Mario Engels and Dennis Diekmier rounded the off the scoring to condemn Hamburg to another season in the lower league.
Second division champions Bielefeld and VfB Stuttgart have won automatic promotion, securing first and second place respectively before Sunday's matches.
Paderborn and Fortuna Duesseldorf were relegated from the Bundesliga.