The purpose of this study was to assess the reliability of critical power (CP) and the total amount of work accomplished above CP (W´) across repeated tests using ecologically valid maximal effort time-trials (TT) under laboratory conditions. After an initial incremental exercise test, ten well-trained male triathletes (age: 28.5 ± 4.7 years; body mass: 73.3 ± 7.9 kg; height: 1.80 ± 0.07 m; maximal aerobic power [MAP]: 329 ± 41 W) performed three testing sessions (Familiarization, Test I and Test II) each comprising three TT (12, 7, and 3 min with a passive recovery of 60 min between trials). CP and W´ were determined using a linear regression of power vs. the inverse of time (1/t) (P = W´ ∙ 1/t + CP). A repeated-measures ANOVA was used to detect differences in CP and W´ and reliability was assessed using the intra-class correlation coefficient (ICC) and the coefficient of variation (CoV). CP and W´ values were not significantly different between repeated tests (P = 0.171 and P = 0.078 for CP and W´, respectively). The ICC between Familiarization and Test I was r = 0.86 (CP) and r = 0.58 (W´) and between Tests I and II it was r = 0.94 (CP) and r = 0.95 (W´). The CoV notably decreased from 4.1% to 2.6% and from 25.3% to 8.2% for CP and W´, respectively. Despite the non-significant differences for both parameter estimates between Familiarization, Test I, and Test II, ICC and CoV values improved notably after the familiarization trial. Our novel findings indicate that for both, CP and W´ a familiarization trial increased reliability. It is therefore advisable to familiarize well-trained athletes when determining the power-duration relationship using TT under laboratory conditions.