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The 1999–2000 KNVB Cup (at the time known as the Amstel Cup for sponsorship reasons) was the 82nd edition of the Dutch national football annual knockout tournament for the KNVB Cup. 86 teams contested, beginning on 31 July 1999 and at the final on 21 May 2000.
All 18 participants of the Eredivisie 1999–2000: six teams entered in the round of 16 of the knock-out stage; one team entered in the first round of the knock-out stage and the rest of the teams entered in the group stage.
The matches of the first round were played on September 22 and 23, 1999. sc Heerenveen entered the tournament this round, during the group stage they were still active in the Intertoto Cup.
The matches of the round of 16 were played on January 27, 28 and 30, 2000. The Eredivisie teams that had been playing in European competitions after qualification last season, entered the tournament this round.