The aim of the study was to analyze the rowing ultra-endurance race where the 100 km Mixed Tandem World Record was broken. The team consisted of one female athlete (age: 32 years; body mass: 61.5 kg; height: 174 cm) who won the 2016 World Championship and one male athlete (age: 40 years; body mass: 74 kg; height: 179 cm), third classified of the same competition. Heart Rate (HR) was monitored with two heart rate sensors Polar H7 Bluetooth Smart and lactic acid was measured with Lactate Pro TM2 analyzer. The z Test was calculated for comparison of proportions based on the quotient that results from dividing effects errors. To analyze HR response, six different HR zones were determined as a function of each player's maximal HR (HR max); Z1: <60%; Z2: 61-70%; Z3: 71-80%; Z4: 81-90%; Z5: 91-95%, and Z6: >95%. Three different exercise intensities were defined as percentages of HR max : low intensity: <70%, moderate intensity: 70-90% and high intensity: >90%. Athletes complete the race in 6:20:52.9 (hh:mm:ss.d) performing alternating relays every two minutes of execution and rest. Results showed that subject 1 spent more time in Z4 (35.56%, p<0.01) and in Z2 (21.26%, p<0.05) and the highest percentage of time was at moderate intensity (51.85%, p<0.05). Predominant work zones of subject 2 were Z1 (19.64%, p<0.05), Z4 (21.38%, p<0.05) and Z5 (22.15%, p<0.05) and the highest percentage of time was at high intensity (42.27%, p<0.001). Lactate concentration of subject 1 and 2 reached a mean of 3.9 ± 5.1 and 5.1 ± 1.3 mmol/l mmol/l, respectively. These findings can help in the preparation of future ultra-endurance races and in the planning of strategies when this type of tests are for relays.