This study examined physiological and race pace characteristics of medium- (finish time < 240 min) and low-level (finish time > 240 min) recreational runners who participated in a challenging marathon route with rolling hills, the Athens Authentic Marathon. Fifteen athletes (age: 42 ± 7 years) performed an incremental test, three to nine days before the 2018 Athens Marathon, to determine maximal oxygen uptake (VO
2 max), maximal aerobic velocity (MAV), energy cost of running (ECr) and lactate threshold velocity (vLTh), and were analyzed for their pacing during the race. Moderate- (
n = 8) compared with low-level (
n = 7) runners had higher (
p < 0.05) VO
2 max (55.6 ± 3.6 vs. 48.9 ± 4.8 mL·kg
−1·min
−1), MAV (16.5 ± 0.7 vs. 14.4 ± 1.2 km·h
−1) and vLTh (11.6 ± 0.8 vs. 9.2 ± 0.7 km·h
−1) and lower ECr at 10 km/h (1.137 ± 0.096 vs. 1.232 ± 0.068 kcal·kg
−1·km
−1). Medium-level runners ran the marathon at a higher percentage of vLTh (105.1 ± 4.7 vs. 93.8 ± 6.2%) and VO
2 max (79.7 ± 7.7 vs. 68.8 ± 5.7%). Low-level runners ran at a lower percentage (
p < 0.05) of their vLTh in the 21.1–30 km (total ascent/decent: 122 m/5 m) and the 30–42.195 km (total ascent/decent: 32 m/155 m) splits. Moderate-level runners are less affected in their pacing than low-level runners during a marathon route with rolling hills. This could be due to superior physiological characteristics such as VO
2 max, ECr, vLTh and fractional utilization of VO
2 max. A marathon race pace strategy should be selected individually according to each athlete’s level.
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