Proposal: Physiological and performance responses to super-oxygenated water
ingested during vigorous exercise
Craig A. Horswill, PhD
Department of Kinesiology & Nutrition
University of Illinois Chicago
Specific Aims and Hypotheses
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Examine the effects of ingesting superoxygenated water on subsequent physical
performance. The effects will be compared with those of ingesting water high in
vitamin C.
Hypothesis: Ingestion of superoxygenated water will enhance performance more so
than the effects of ingesting vitamin C water.
Examine the effects of ingesting superoxygenated water on the redox state of the
blood compared to that of ingesting water containing vitamin C.
Hypothesis: Ingesting superoxygenated water will result in greater levels of reactive
oxygen species as indicated by the ratio of reduced to oxidized glutathione.
Background
Ingestion of superoxygenated water has been identified as a potential adjunct to
providing oxygen to the blood. Prior research, though, lacks supportive findings for
performance enhancement. Four studies have been published in which subjects
performed progressively workload tests that culminated in maximum effort, i.e. VO 2 max
(Willment et al 2002; Wing-Gaia et al 2005; Leibetseder et al 2006; McNaughton et al
2007). Briefly, VO2 max did not increase in subjects of mixed fitness level. Only one of
these studies included subjects that could be considered highly fit (average VO 2 max of
61.9 mL/kg/min) and the performance testing was done at submaximal intensity, which
is unlikely to benefit from surplus oxygen (Sommer 2013).
One explanation is that one determinant of VO2 max – pulmonary function – has seldom
been demonstrated as a limiting factor in maximal oxygen consumption and the
associated performance (Bassett and Howley 2000). As an exception, though, Powers
et al showed that in very high-level endurance athletes, the pulmonary system may be a
limiting factor. By giving the high-end athletes 26% oxygen to breath during a VO2 max
test, the subjects increased their VO2 max by 6.6% (+4.6 mL/kg/min). Breathing the
higher percentage of oxygen prevented arterial desaturation of the blood (SaO2 of 95.5
+0.4% vs 90.6 +0.8 % when breathing normal room air of 20.9% at max effort). The
same treatment had absolutely no effect on the control group, which was fit but not
highly trained (VO2 max of ~57 mL/kg/min). The authors concluded that the pulmonary
system may limit oxygenation and prevent desaturation at max (Powers et al 1989) in
certain individuals. Amman has speculated that sluggish ventilatory response,
anatomical limitations, competition between locomotor muscles and respiratory muscles
for blood flow, and intrathoracic pressure effects on cardiac output could explain how
the pulmonary system limits VO2 max (Amman 2011). While it may be a long shot,
alternative methods to elevating oxygen in the blood of highly trained endurance
athletes might promote an increase in VO2 max.
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Proposal: Physiological and performance responses to super-oxygenated water
ingested during vigorous exercise
Oxygenation of water could influence performance and physiology through another
mechanisms. Oxygenated water has been demonstrated to produce ROS in the water
(Schoenberg et al 2002) and in the body after ingestion at elevation (Wing-Gaia et al
2005). Reactive oxygen species (ROS) are thought to be generated at high rates and
amounts during exercise (Allen et al 2008; Powers and Jackson 2008). Allen et al have
reviewed comprehensively skeletal muscle fatigue and explained that ROS may play a
role in optimizing muscle function as well as contributing to fatigue through several
potential mechanisms (Allen et al 2008). A low level of ROS might actually be beneficial
and contribute to enhanced performance by mechanisms yet to be explained (Allen et al
2008).
A comparison against the opposite end of the redox-state, i.e., that incurred after
ingesting of a high dose of an antioxidant, could help sort out the role of ROS in
optimizing performance. In chronic feeding studies, intake of vitamin C has failed to
enhance VO2 max compared to placebo, and more than likely impairs performance
based on absence of tissue adaptive responses in the animal model (Gomez-Cabrera
2008). The study of ROS in the exercising human body is challenging because of the
limited half-life and inability to measure ROS levels directly. However, glutathione can
been used as an indirect index of changes in ROS concentrations as a result of
exercise and antioxidant treatment (Medved et al 2004; Powers and Jackson 2008).
The purpose of the study is to compare the acute effects of superoxygenated water
ingestion on performance as determined during a running VO2 max during a
progressive workload treadmill test. VO2 max and the final stage (speed and grade) of
running will be used as the primary outcome variables. Blood samples will be taken
before ingestion of the water, one hour after ingestion, and immediately after the VO2
max test ends.
Study Design
For this study we will recruit high-end male endurance athletes (treadmill VO2 max >65
mL/kg/min) between the ages of 18 and 35 y. The protocol and consent, will be
developed and submitted for review and approval by the UIC Institutional Review Board
prior to recruitment and testing of subjects.
Once enrolled, subjects will be measured for physical characteristics, body composition
(BodPod, Cosmed, Chicago, IL and/or multi-segmental, multi-frequency electrical
impedance InBody, California), and have VO2 max confirmed with a progressive
workload test (Parvo Medics TrueOne 2400 metabolic measurement system, Park City,
UT).
For testing of effects of superoxgenated water, a double-blind crossover experimental
design will be used. Subjects will serve as their own controls and experience both
treatments. The order of administration of the beverages will be randomized and
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Proposal: Physiological and performance responses to super-oxygenated water
ingested during vigorous exercise
balanced with the 12 subjects (six subjects randomly selected to receive placebo first
and oxygenated water second and six randomly selected for the opposite order). One
week will be allowed between tests for the washout period.
Subjects will keep a record of food and fluids consumed during the 24-h period before
the test date. For the second trial, subjects will receive a copy of the record and told to
replicate it during the 24-h period before the second test.
On the test day, subjects will report to the lab 3 to 4 h postprandial. An initial blood
sample will be drawn for the baseline value of GSH and GSSG. Subjects will then
consume a standardized volume of test beverage and rest for 1 hour. A second blood
will be drawn. After a brief, standardize period to warm up on the treadmill, subjects will
drink another standardized volume of the test beverage, wait 15 min and begin the max
test. A modified Bruce protocol will be used in which the speed will remain constant but
the slope of the treadmill will rise every 3 min. Oxygen consumption, VCO2, heart rate,
final slope and speed achieved, and ratings of perceived exertion will be measured.
The ventilator threshold will be determined from the VO2 and VCO2 data as an index of
anaerobic threshold.
Blood samples will be analyzed for reduced and oxidized glutathione concentrations
(Total Glutathione Assay Kit, Northwest Life Sciences LLC, Portland, OR). The assay
specifically measures total glutathione and components in the red blood cells, which will
serve as a proxy for the working tissues, i.e., skeletal muscle, in this study.
Data will be summarized as the mean and standard deviation for dependent variables
per treatment and time point of measurement. Inferential statistics will consistent of 2way analyses of variance adjusted for repeated measurements. The Newman-Keul
post hoc test will be used to determine were difference occur if appropriate. A
probability of 0.05 is selected as the alpha level for statistical significance.
Role of Student
The student involved in the project will help with recruitment, data collection, lab assays,
data reduction, and helping produce the final report. The student can expect to work
~20 hours a week in June and July for data collection and early August to complete the
report.
Required Student Proficiencies
The student involved in this project should have prior laboratory and research
experience. The student should have a basic understanding of and be able to apply
statistical methods (have taken KN200) and understand respiratory responses to
exercise (completed KN352). In addition, an understanding on muscle physiology and
mechanisms of fatigue is critical (completion of KN441).
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Proposal: Physiological and performance responses to super-oxygenated water
ingested during vigorous exercise
References
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Amann M. Pulmonary system limitations to endurance exercise performance in human. Exp.
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