PURPOSE: A number of physiological diagnostics were developed. However, the timeline-related diagnostics of maximal anaerobic glycolytic capacity remain unclear. The objective of this study was to evaluate the reliability and validity of a sprint running test to assess the anaerobic capacity.METHODS: The study was divided into three parts. Sixty-one male (24±4 years, 181.0±4.3 cm; 78.5±5.9 kg) and twelve female (25±3 years, 167.0±0.6 cm, 60.4±5.7 kg) sports students participated in this study. Twenty-five subjects (13 males, 24±2 years, 181.0±0.5 cm, 78.5±5.9 kg; 12 females, 25±3 years, 167.0±0.6 cm, 60.4±5.7 kg) performed incremental step tests at running track and several linear sprints on a running track (LSRT) with different time durations (8, 10, 12, and 14 seconds)(part I) on different days. Twenty-five male subjects (24±3 years, 180.7±6.7 cm, 84.6±8.8 kg) conducted a 10 or 12 second sprint running on a non-motorized treadmill (NMT)(part II). In part III, twenty-three male subjects (24±2 years, 181.4±5.8 cm, 74.5±7.4 kg) ran a 10 second LSRT and NMT on consecutive days. Capillary blood samplings were taken before (Lac<sub>r</sub>) and after the sprint running for ten minutes at one minute intervals to find out maximal lactate concentration after exercise and to calculate the maximum lactate production rate (LPR<sub>max</sub>).RESULTS: For all parts reliability for LPR<sub>max</sub> was proven (Part I: 8 seconds: ICC: <i>r</i>=.89; 10 seconds: ICC: <i>r</i>=.82; 12 seconds: ICC: <i>r</i>=.92; 14 seconds: <i>r</i>=.84, respectively; Part II: 10 seconds: ICC: <i>r</i>=.76; 12 seconds: ICC: <i>r</i>=.79). To analyze validity for LPR<sub>max</sub>, Part III was conducted and proven valid (ICC: <i>r</i>=.96, p=.074).CONCLUSIONS: We demonstrate that LSRT and NMT reliably determine anaerobic capacity and can be used as a valid tool for physiological performance diagnostics.