Matthijs de Ligt will watch from the sidelines as both the Netherlands and Ukraine aim to end miserable runs in their Euro 2020 opener on Sunday.
De Ligt missed the Oranje's final warm-up friendly due to a groin issue and will not recover in time to face Ukraine in Amsterdam.
Coach Frank de Boer said: "The game comes a little too early. We don't want to take any chances. We've got two more group games.
"If it had been very important, he could have played. He's very close to it."
It is a blow to a Netherlands side who have not won at the European Championship since the 2008 group stage.
The Dutch were quickly beaten in the knockout phase of that competition, lost all three matches at Euro 2012 and then failed to qualify four years later.
Centre-back is suddenly a potential area of weakness, with Virgil van Dijk missing the tournament, while De Boer only confirmed the identity of his starting goalkeeper – 38-year-old Maarten Stekelenburg – on Saturday.
Regular number one Jasper Cillessen, cut from the squad following a positive coronavirus test, was the only ever-present in qualifying besides De Ligt.
Ukraine might make for accommodating opponents for Stekelenburg, though, having themselves lost five in a row at the Euros and failed to score in each defeat.
Andrey Shevchenko, now their coach, is the only player to have ever scored for Ukraine at the finals, as they have failed with 67 shots since his 2012 brace against Sweden.
PLAYERS TO WATCH
Netherlands – Memphis Depay
Even with De Boer switching to an unpopular 5-3-2 formation, Depay will be the chief threat in attack. Although Georginio Wijnaldum (eight) outscored his team-mate in qualifying, Depay was involved in a goal every 38 minutes (six goals and seven assists in 495 minutes) – the best rate of any player involved in five or more.
Ukraine – Andriy Pyatov
If Depay is as involved as the Netherlands hope, that will mean another busy outing for Pyatov. The Ukraine goalkeeper saved 88.6 per cent of the shots he faced in qualifying (31 of 35), which was the best rate of any keeper to appear in six or more games.
KEY OPTA FACTS
– This is the first meeting between the Netherlands and Ukraine at a major tournament (World Cup or European Championship). The Dutch are unbeaten in their two friendly encounters – winning 3-0 in Rotterdam in May 2008 and drawing 1-1 in Donetsk in August 2010.
– The Netherlands are playing their first major tournament game at the Johan Cruyff Arena since the Euro 2000 semi-final against Italy, a game in which the Dutch failed to score despite having 31 shots and 71 per cent possession (0-0 after extra time, 1-3 on penalties).
– Since and including 1988, only Germany (six) have reached the semi-finals of the European Championship more often than the Netherlands (four). However, the Netherlands have not made the final four since 2004 and they have not won a game in the tournament since 2008.
– After winning their first game at the European Championship (2-1 v Sweden, 11 June 2012), Ukraine have subsequently lost their past five. In fact, only Yugoslavia have had a longer losing run in the history of the tournament (six in a row, from Euro 1968 to Euro 1984).