There were unhappy scenes at Volksparkstadion as the only club that had enjoyed the distinction of being the only club to have played in every season of the Bundesliga since its establishment in 1963 was relegated.
Hamburg were relegated from the Bundesliga for the first time in their history on Saturday, as a 2-1 win over Borussia Moenchengladbach proved too little too late.
A proud run of 55 season in Germany's top flight came to an ugly end as Hamburg fans set off a flurry of fireworks, releasing a cloud of black smoke across the Volksparkstadion, forcing play to be suspended just minutes from time.
"This is an enormously painful moment for everyone at the club, which has lost one of its defining characteristics," said Hamburg president Bernd Hoffmann.
The six-time German champions, who won the European Cup in 1983, are the only team to have played in every Bundesliga season since the league's inauguration in 1963, earning them the nickname "dinosaur".
Although Hamburg battled to victory over Moenchengladbach, they were doomed to relegation as Wolfsburg thrashed Cologne to finish third from bottom. Wolfsburg will go into a relegation playoff with Holstein Kiel, who finished third in Germany's second tier.
The day started disastrously for Hamburg, as Joshua Guilavogui fired Wolfsburg ahead in the opening minute against Cologne.
In Hamburg, Aaron Hunt put the home side in front before Josip Drmic equalised for 'Gladbach.
Jonas Hector brought Cologne level just after the half-hour mark with an exquisite chip, but Divock Origi, on loan from Liverpool, restored the lead for Wolfsburg and edged Hamburg closer to disaster.
Lewis Holtby's curling strike kept Hamburg's hopes alive, but when Bobby Wood was sent off 20 minutes from time, their fate seemed sealed.
Minutes later, Robin Knoche headed in Wolfsburg's third, and when Josip Brekalo added a fourth in stoppage time, Hamburg's fate was sealed.
After a 15-minute delay due to crowd trouble, the game briefly resumed. The famous digital clock in the Volksparkstadion, which displays how long the club have been in the top flight, must now finally be switched off.
"I am so disappointed," said Hamburg coach Christian Titz. "I was convinced that we could win the game today, and that it would be enough to stay up. The team and most of the fans gave us a worthy send-off from the top flight."
At the other end of the table, Hoffenheim pipped Bayer Leverkusen and RB Leipzig to a Champions League place next season as they beat Borussia Dortmund 3-1 to third place.
Dortmund goalkeeper Roman Buerki gifted Hoffenheim possession with a dreadful clearance on 26 minutes, allowing Andrej Kramaric to fire the home side in front.
Marco Reus equalised for Dortmund after half-time, but goals from Adam Szalai and Pavel Kaderabek fired Hoffenheim into Europe's elite.
Lucas Alario grabbed a brace as Leverkusen eased to a 3-2 win over Hanover, as both they and Leipzig secured a Europa League place.
Two strikes from Jean-Kevin Augustin and goals from Timo Werner, Dayot Upamecano and Ademola Lookman helped Leipzig to a frantic 6-2 win over Hertha Berlin.
Shock defeat for Bayern
For Bayern Munich, who wrapped up the title last month, Saturday was supposed to be a day of celebration, as they were presented with the Bundesliga trophy for the sixth season in a row.
Bayern, though, were unable to give coach Jupp Heynckes an appropriate send-off in his final home game, as they stumbled to a shock 4-1 defeat to Stuttgart.
Ruthless on the break, the visitors tore through Bayern, as Daniel Ginczek, Anastasios Donis and Chadrac Akolo all found the net on the counter-attack.
"The defeat was frustrating, but we still have every reason to celebrate," said Bayern goalkeeper Sven Ulreich.
Bayern celebrated with the customary on-field showers of confetti and beer, as injured players Arturo Vidal, Manuel Neuer and Jerome Boateng joined their team-mates on the pitch.
Heynckes was spared the traditional beer shower for the coach, as he disappeared down the tunnel. The 72-year-old will return to retirement after next weekend's German Cup Final against Eintracht Frankfurt.
"We wanted to send Heynckes off with a win, but we have another chance to do so next week," said defender Niklas Suele.