This study examined the association between implementation of the revised stoppage-time regulation following the 2022 FIFA World Cup and the temporal and locomotor activity profile of professional football using multilevel hierarchical modeling. A total of 312 matches (624 team-level observations) from the 2022–2023 Turkish Super League season were analyzed, comparing prechange (Matchweeks 1–14) and postchange (Matchweeks 15–38) periods using linear mixed-effects models (LMM), with match location as a fixed covariate and team clustering as a random effect. Total match duration (100.10 ± 3.11 vs. 103.02 ± 4.07 min;
F
(1, 622)
= 102.33,
p
< 0.001,
d
= 0.79), ball-in-play time (
F
(1, 622)
= 44.97,
p
< 0.001,
d
= 0.54), and offensive/defensive phase durations (
F
(1, 622)
= 6.64,
p
= 0.010,
d
= 0.20) increased significantly post-change. Total distance (
F
(1, 622)
= 105.80,
p
< 0.001,
d
= 0.79), moderate-intensity (
F
(1, 622)
= 46.80,
p
< 0.001,
d
= 0.51), high-intensity (
F
(1, 622)
= 12.39,
p
< 0.001,
d
= 0.28), and sprinting distances (
F
(1, 622)
= 21.90,
p
< 0.001,
d
= 0.35) also increased. Time-normalized total distance (
F
(1, 622)
= 6.21,
p
= 0.013,
d
= 0.21), moderate-intensity (
F
(
1, 622)
= 11.45,
p
< 0.001,
d
= 0.28), and sprinting distances (
F
(1, 622)
= 7.85,
p
= 0.005,
d
= 0.22) remained significantly higher, whereas relative high-intensity running was unchanged (
F
(1, 622)
= 1.04,
p
= 0.308,
d
= 0.08). The revised regulation was associated with greater match volume and physical demands, including higher sprinting density. In conclusion, while the updated stoppage-time regulation effectively increases effective playing time, it imposes a heightened physical load—particularly elevated maximal-velocity density—requiring practitioners to recalibrate weekly periodization and late-match substitution strategies.