Former England midfielder Gareth Barry played for West Brom's Under-21s at the age of 37 in the Checkatrade Trophy but failed to save them from elimination.
Barry, who holds the record for most Premier League appearances, played 45 minutes on his return from injury as Albion lost 2-1 at Macclesfield, the EFL's bottom side.
Meanwhile, Swansea's Under-21s beat Charlton to move into the second round.
Portsmouth beat Tottenham's youngsters 3-2, with both sides progressing.
Spurs and Swansea are two of seven clubs with Category One academies to confirm their place in the final 32, with Southampton capable of joining them on Wednesday if they beat Southend by three or more goals.
For Barry, a Premier League and FA Cup winner with Manchester City, the group game at Moss Rose was his EFL Trophy debut.
He started on his comeback from a calf injury that kept him out for five weeks and lined up alongside former Wales goalkeeper Boaz Myhill against the managerless Silkmen, who finished second in the Northern Section's Group C to go through.
While traditionally a knockout tournament for League One and League Two clubs, the competition changed format in 2016 to accommodate 16 academy teams from the Championship and Premier League.
Stoke Under-21s beat Carlisle United on 5-4 on penalties after drawing 1-1 at Brunton Park, and go into Friday's draw as Northern Group A runners-up behind Sunderland, who had a following of almost 1,600 in the crowd of 2,377 for their 1-0 win at Morecambe.
Shrewsbury, who sacked manager John Askey on Monday, won 2-1 at Crewe to reach the second round.
Victory won Northern Group D for the Shrews as they moved above Manchester City's Under-21s, who had already qualified.
Newport overcame Plymouth 2-0 to reach the knockout stages, edging out Swindon on goal difference in Southern Group C.
Coventry City, the 2017 Trophy winners, were knocked out after being beaten 2-0 at Cheltenham.
The draw for round two will take place from 13:00 GMT on Friday 16 November.