1. Pilot Study “ Effect of Sports Oxyshot on Selected Indices of Endurance Performance in Trained Athletes ” Ian Gillam PhD Exercise Physiologist and Nutritionist AFL Melbourne Demons FC Assoc Prof Steve Selig PhD Exercise Physiologist, Victoria University

2. Aims of Pilot Study  Effect of Sports Oxyshot on Indices of Aerobic Performance  Maximal Performance indices  Peak VO2  Peak Power  Sub-maximal Aerobic Performance indices @ 4 mM Lactate Threshold  Oxygen Uptake  Power  Heart Rate

3. Study Design of Trial  Double Blind, Placebo-Controlled, Cross-Over Design  Supplementation period of one week each treatment  Either 15 ml of Sports Oxyshot (OS) or Placebo (PL)*  Ingested each morning for 7 days prior to each exercise test  Treatment Compliance was 100% for all subjects  Subjects Paired based on age (but not gender) and training load and treatment randomly assigned  * Degassed tonic water

4. Study Design of Trial 2  Identical training in 24 hours prior to each exercise test and no training 12 hours prior to test  Diet was controlled in 24 hours prior to each test  Each subject undertook each test at same time of day  All tests conducted under standard Laboratory conditions

5. Subjects  Four triathletes  2 Masters Ironman triathletes  DG (F, 40 years)  MB (F, 33 years)  Due to Viral infection in the few week before testing data has not been included in analysis  Training of 16-20 hours/week (Bike, Swim, Run)  2 Olympic Distance triathletes  CH (M, 23 years). Ranked in top 10 U23 triathletes in Victoria  NW (F, 26 years). Ranked 5th as a road cyclist in Victoria  Training 14-18 hours/week (Bike, Swim, Run)

6. Exercise Testing  Conducted on Electromagnetic Cycle Ergometer  Cycle set-up standardized for each subject according to their preferred seat and frame dimensions  Protocol for Progressive Exercise test  3 min warm up at 50 watts  Start test at 100 watts, with an increase of 25 watts each 2 mins  Measurements made in the last 30 secs of each Workload  Crank rpm selected by each subject recorded at each minute  Power outputs increased until volitional exhaustion or subject was unable to maintain a crank rpm @ 60 revs.

7. Parameters Measured  Cycle Power output and crank rpm  Metabolic Measurements  Oxygen uptake, VE, RER each 30 secs (Med Graphics)  HR Via ECG telemetry  Capillarized Whole blood Lactate  Via Finger prick during the last 15 secs of each power output  Analysed by Lactate Pro Auto-analyser  Borg RPE (6-20 scale) 

8. Data Analysis  Maximal Data  VO2 max and Peak Power Compared for OS and PL trials  Determination of 4 mM HLa Threshold  “Best Fit” Graphs drawn for the following :  Power Output VS HLa  VO2 Vs HLa  HR Vs HLa

9. Exercise Testing  Conducted on Electromagnetic Cycle Ergometer  Cycle set-up standardized for each subject according to their preferred seat and frame dimensions  Protocol for Progressive Exercise test  3 min warm up at 50 watts  Start test at 100 watts, with an increase of 25 watts each 2 mins  Measurements made in the last 30 secs of each Workload  Crank rpm selected by each subject recorded at each minute  Power outputs increased until volitional exhaustion or subject was unable to maintain a crank rpm @ 60 revs.

10. Maximal Power Outputs SubjectPL(Watts)OS(Watts) Diff (%) CH389394+1.2% DG311303-2.6% NW356368 + 3.3%

11. Maximal Oxygen Uptake SubjectPL(ml/kg/min)OS(ml/kg/min) Diff (%) CH76.277.2 + 1.3% DG58.357.3-2.3% NW62.964.73.3%

12. Power Outputs @ 4 mM Threshold SubjectPL(Watts)OS(Watts) Diff (%) CH283293+3.5% DG175179+2.2% NW285305 + 7.0%

13. VO2 BTPS @ 4 mM Threshold SubjectPL(ml/min)OS(ml/min) Diff (%) CH37003925 + 6.1% DG34903620 + 3.7% NW42503980 - 6.8%

14. Power Vs HLa CH HLa (mM) Power (Watts)283 293 OS PL

15. VO2 vs HLa VO2 ATPS (ml/min) 36003400 HLa (mM) OS PL

16. HR vs HLa HLa (mM) 170 162 HR (bpm) OS PL

17. Power Vs HLa DG HLa (mM) 275260Power (Watts) OS PL

18. VO2 Vs HLa DG VO2 ATPS (L/min) HLa (mM) 3.19 3.32 OS PL

19. HR vs HLa HLa (mM) HR (bpm) DG 179 175 OS PL

20. Summary of Results  While data are extremely limited due to small sample size  No effect of OS on maximal performance data  Peak Power or VO2 max  OS increased the 4 mM HLa threshold by 2.2- 7.0% based on VO2 and Peak Power data  On an individual basis, this could result in a 2-3 minute/hour performance improvement when completing a prolonged endurance event such as a triathalon

21. How might OS improve endurance performance 1?  OS contains 17% stabilized Oxygen in a strong hypochlorite solution  15 ml dose contains only 2-3 ml, so the additional oxygen supplied in OS cannot responsible!  As a 5% OS solution has been shown to increase arterial pO2 by 5-10% in healthy human subjects for 30-120 mins after ingestion  This clearly shows OS is not only bio-available  But also results in significant increases in blood PO2 changes

22. How might OS improve endurance performance 2?  Could the increase in pO2 mediate secondary changes?  Could this alter the regulation of respiratory or cardiac response by arterial chemoreceptors?  UNLIKELY- no evidence of difference in VE during exercise during this study  Alterations in levels of red cell 2,3 DPG and increased oxygen delivery to the active tissues?  POSSIBLE. Chronic changes in arterial PO2 regulate DPG Levels  Evidence of an “induction period” during chronic use of OS

23. Future Research Studies  1. Increase sample size of current study to determine significance of the effect on parameters of endurance performance in athletes.  2. Examine effect of long term use of OS on arterial pO2, red cell 2,3 DPG, indices of oxidative damage and endurance performance  3. Determine the Oxygen species in OS by electron spin resonance studies and specifically assess if there is any evidence of transient reactive oxygen species present.  4. Examine the potential use of OS to increase oxygen delivery to the tissues when on long haul flights and to assist recovery in athletes