Anel Ahmedhodzic became the first player to not wear the rainbow armband in seven years during Sheffield United's loss to Liverpool on Wednesday night.
The Blades suffered their first defeat in Chris Wilder's second spell as manager, with Virgil van Dijk and Dominik Szoboszlai getting on the scoresheet for Jurgen Klopp's side.
Ahmedhodzic was given captain duties in the absence of John Egan and Chris Basham but did not wear the rainbow armband, opting for the standard one instead.
All 20 Premier League captains are required to wear an armband featuring traditional Pride colours in allocated matches as part of the Rainbow Laces campaign.
The Premier League's official website stated: 'Our clubs will come together between 2-7 December to celebrate Stonewall's 2023/24 Rainbow Laces campaign and show support for all LGBT people in football and beyond.'
Anel Ahmedhodzic became the first player to not wear the rainbow armband in seven years
The Rainbow Laces campaign requires Premier League captains wear an armband featuring traditional Pride colours
Liverpool captain Van Dijk wore the armband as did Arsenal's Martin Odegaard and Manchester City's Kyle Walker during the midweek fixtures.
When told Ahmedhodzic didn't wear a rainbow armband, Wilder stated that he was unaware of the situation.
'I didn't know,' he said in a press conference.
'It's not a club decision or a personal decision. I just didn't know he did that.'
Meanwhile, Sheffield United picked up just their second win of the season on Saturday after beating Brentford 1-0 thanks to James McAtee's strike.
The Blades will seek to make it back-to-back wins for the first time this term when they take on Chelsea next weekend.
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