Background: Research into key performance factors in trail running, particularly in vertical kilometer (VK) races, is crucial for effective training and periodization. However, recent studies on metabolic and cardiorespiratory responses during VK races, especially using field tests, are limited. Objectives: Therefore, the aim of this study is to evaluate the metabolic and cardiorespiratory responses during a VK field test, identifying differences based on sex and performance level, as well as key performance factors and their deterioration due to fatigue. Fifteen trained trail runners (ten males and five females, 19 to 38 years old) perform a VK race. Methods: The global physiological response is evaluated using the portable gas analyzer Cosmed K5 and the local response using near-infrared spectroscopy technology. Results: In gender comparisons, the ANCOVA test shows significant differences (p < 0.05) in the ventilation, tidal volume, expiratory time-to-inspiratory time ratio, inspiratory flow rate, end-tidal CO2 partial pressure, heart rate, oxygen pulse, and total hemoglobin. Additionally, the performance comparison reveals significant differences in the variables’ velocity, oxygen consumption, carbon dioxide production, ventilation, dead space-to-tidal volume ratio, total time of the breathing cycle, expiratory time-to-inspiratory time ratio, inspiratory duty cycle, expiratory fractions of CO2, quadriceps saturation index, and VE/VCO2 ratio. Finally, the correlation analysis shows oxygen consumption (r = −0.80 mean; r = −0.72 peak), carbon dioxide production (r = −0.91 mean; r = −0.75 peak), expiratory time-to-inspiratory time ratio (r = 0.68 peak), ventilation (r = −0.58 mean), and quadriceps saturation index (r = 0.54 mean; r = −0.76 coefficient of variation) as the key performance factors in the VK race. Conclusions: Overall, the physiological analysis indicates the importance of local muscular adaptations and respiratory system capacity in this type of short-duration race.
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