Jordan Pickford’s England were edged out 1-0 by Belgium as Roberto Martinez’s side secured top spot in World Cup Group G.
The result means that the Three Lions will face Group H winners Colombia in the last 16 on Tuesday night, with Belgium setting up a clash with Japan on Monday.
With qualification to the knockout stages already secured, Gareth Southgate made eight changes to the team that hammered Panama 6-1 on Sunday.
Only Pickford, former Blues defender John Stones and midfielder Ruben Loftus-Cheek retained their places from the thumping win in Nizhny Novgorod.
After the sides went in at half-time on level terms, Belgium broke the deadlock on 51 minutes. Adnan Januzaj showed some nifty footwork to work some space just inside the penalty box before rifling a left-footed shot into the top corner.
England responded well to the setback and were close to finding an equaliser on 65 minutes. After a neat link-up with Jamie Vardy, Rashford went through one-on-one with Courtois but the Chelsea ‘keeper made a fingertip stop to preserve Belgium’s advantage.
England continued to threaten but failed to carve out any clear-cut opportunities as Belgium saw out the victory.
Next up for Pickford – who is now the only England player to play every minute in the tournament – is a round of 16 showdown with Colombia on Tuesday night in Moscow (kick-off 7pm).