The aim of the study was to test the hypothesis if supplemented NADH is able to enhance the available cellular energy prior and during physical exercise as well as enhance after exertion on highly conditioned athletes.

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Enadalert Enhance Intracellular Energy Levels in Trained Athletes

Objective:
The aim of the study was to test the hypothesis if supplemented NADH is able to enhance the available cellular energy prior and during physical exercise as well as enhance after exertion on highly conditioned athletes.

Methods:
Test subjects

14 highly conditioned endurance athletes, who had a significant higher aerobic performance compared to normal healthy individuals (VO2max > 55mL/kg/min), were included in a double blind cross over placebo (Nicotinamide) controlled trail. The test intervals were 4 weeks ingestion of 30mg NADH (ENADAlert) or Nicotinamide daily, interrupted by a 6 weeks wash out period. On the test days no NADH/placebo intake was allowed.

Extracellular NADH metabolization assay (ENMA)

500 ml of EDTA-blood (used between one and four hours after collection) are diluted with 1xPBS (8 g/l NaCl; 0.2 g/l KH2PO4; 1.15 g/l Na2HPO4; adjusted to pH 8.0 with 6M KOH) in a Centrisart 1 filter tube with a cut off range of 20,000 MW (Sartorius), 50 ml of NADH solution (8 mg NADH/ml 1xPBS) are added to start the reaction. This mixture is incubated for two hours at 37°C. The reaction is topped by centrifuging the tube for 10 minutes. The filtrate which contains the not consumed NADH is collected and analyzed by HPLC. Two Standards with 2 ml 1xPBS including a) 25 ml or b) 50 ml of NADH solution (described above) incubated under the same conditions are used to quantify the metabolized NADH amount.

Determination of NADH with HPL
The determination is in principle according to the method of Formato et al. [1] and described in detail in Nadlinger et al. [2] in brief: A Shimadtzu LC 10A System was used with a Lichrospher RP18, 5 mm 250×4 mm (Merck) column. NADH was measured at 340 nm with a diodearray detector (Shimadzu SPD-M10A).

The result of the assay was calculated as mg of NADH not metabolized by 10 ml blood compared to the 50 ml NADH standard (8mg NADH, 0mg NADH metabolized).

Results:
Both groups show a lower NADH metabolization after administration of 30mg ENADAlert compared to the administrated placebo. This effect is found for both groups at all analyzed times (before exercise, after warm up, after maximum aerobic performance and on the next day).

Conclusions:

Although there was no NADH (ENADAlert) administrated on the day of testing, NADH metabolization was less pronounced after the ENADAlert intervals (higher residual NADH values in the ENMA) than after the placebo intervals. Considering that the ENMA correlates with the intracellular ATP/ADP ratio, we conclude that the intake of NADH leads to a higher amount of available cellular energy.

REFERENCES:

[1] M. Formato, B. Masala, G. DeLuca, The levels of Adenine Nucleotides and Pyridine Coenzymes in red blood cells from the newborn, determined simultaneously by HPLC, Clin. Chim, Acta 189 (1990) 131-138.

[2] K. Nadlinger, W. Westerthaler, D. Storga-Tomic, J.G.D. Birkmayer, Extracellular metabolization of NADH by blood cells correlates with intracellular ATP levels, Biochem. et Biophys Acta, 2002 (in press).