Endurance athletes rely on an efficient balance of oxygen delivery and uptake to sustain high–intensity exercise and enhance their athletic performance1,2. The maximal oxygen uptake test (VO2-max) is a vital physiological assessment that determines sports performance by analyzing exhaled–gases and cardiorespiratory variables during incremental exercise intensity1. This assessment, known as ergospirometry or Cardiopulmonary Exercise Testing (CPET), reflects the exercise response of the cardiovascular, respiratory, and muscular systems3. Along these lines, the increased energy cost associated with breathing, referred to as the cost of breathing (COB), heightens the demand for nutrients and oxygen in the surrounding tissues. This phenomenon has been documented to potentially reduce blood flow to the muscles involved in active movements, resulting in a decreased tolerance to physical exertion and an early cessation of exercise progression due to the metabolic reflex4.
During a VO2–max test, it is also possible to identify the ventilatory thresholds (VTs), which correspond to specific exercise intensities marking the transition from aerobic to anaerobic metabolism (aerobic threshold or ventilatory threshold 1 [VT1], and anaerobic threshold or respiratory compensation point [RCP] or ventilatory threshold 2 [VT2])5. The VTs reflect the ventilatory responses that compensate for metabolic changes during incremental exercise6. By identifying these thresholds, CPET offers a comprehensive evaluation by integrating the responses of multiple biological systems critically engaged during high–intensity exercise.
However, while ergospirometry is widely considered the gold standard for evaluating CPET, it does not capture metabolic changes occurring at the muscle level. These changes are crucial for understanding the physiological limiting factors associated with the lack of progression during high–intensity exercise in endurance athletes. In this context, NIRS technology has emerged as a valuable tool in exercise science, aiding in analyzing hemodynamic variables at the microvascular muscle level7.
In recent years, sports professionals and researchers have used a wide range of commercial wearables equipped with NIRS technology to explore non–invasive muscular changes during exercise, providing the ability to determine VT1 and VT2 with this technology8. Thus, the integrative analysis of data from NIRS and CPET offers a comprehensive understanding of physiological responses to exercise.
The NIRS technology utilizes the modified Beer-Lambert law to quantify changes (D) in the concentrations of oxyhemoglobin (O2-Hb) and deoxyhemoglobin (H-Hb) during exercise7. At the local tissue level, a decrease in O2-Hb reflects an increase in local metabolic demand, while an increase in H-Hb reflects an increase in oxygen extraction. Total hemoglobin (tHb), the sum of O2-Hb and H-Hb, is used as an index of local tissue blood flow. Conversely, the difference between O2-Hb and H-Hb (Hbdiff) provides an index of tissue oxygen extraction9. The tissular saturation index (TSI), calculated as the ratio of O2-Hb to tHb, reflects the tissue oxygen saturation level and indicates the balance between local oxygen delivery and uptake10,11. Thus, NIRS data give critical insights into the physiological status at the microvascular level, providing a detailed understanding of tissue oxygenation and hemodynamics that complements the information obtained from CPET.
This detailed understanding provided by NIRS technology extends to many practical applications. Recent research highlights the versatility of NIRS and demonstrates its practical application in monitoring respiratory12,13 and locomotor muscles7, as well as brain regions involved in motor act ideation, such as the prefrontal cortex (PFC)14,15. This broad applicability underscores the ability of NIRS to provide comprehensive insight into physiological responses to various types of muscle contractions (concentric or, eccentric, or isometric contractions) and exercise.
By analyzing exercise-induced DTSI at both the muscular and cerebral levels, NIRS provides a valuable potential for identifying associations between peripheral and central limiting factors that affect the progression of exercise16,17. For example, among central limiting factors, decreased blood flow resulting from cerebral vasoconstriction caused by compensatory hyperventilation due to elevated hydrogen levels from anaerobic metabolism and increased blood lactate during high-intensity exercise is a significant contributor to the reduction in TSI in the prefrontal cortex17,18. In contrast, peripheral limiting factors are characterized by an imbalance between oxygen supply and demand in the exercising musculature19. Reduced local oxygen delivery and increased oxygen consumption can lead to tissue deoxygenation, as evidenced by decreased TSI20. This distinction highlights the multifaceted nature of performance limitations during high–intensity exercise, where both central and peripheral mechanisms are critical. This understanding suggests that delaying the onset of these limiting factors during exercise may contribute to improved athletic performance.
To fully leverage the potential of NIRS technology in identifying these limitations, standardized procedures are essential to ensure high-quality data collection and analysis. This document outlines methods for conducting maximal endurance exercise testing using NIRS technology to collect physiological data and elucidate the relationship between central and peripheral limiting factors during high-intensity exercise in endurance athletes. The proposed protocol provides a standardized approach to ensure consistency and accuracy in assessing the physiological phenomena underlying these limiting factors.