Effects of Post-Exercise Recovery Drink Composition on Subsequent Performance in Masters Class Athletes
Phase: NA
Conditions: Dietary Supplementations
Sponsor: Erica Goldstein

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PROTOCOL TITLE: 
Effects of Post-Exercise Recovery Drink Composition on Subsequent 
Performance in Masters Class Athletes  
PRINCIPAL INVESTIGATOR: 
Erica R. Goldstein, MS, RDN 
Division of Kinesiology  
561-324-6955 
egoldstein@knights.ucf.edu 
VERSION NUMBER/DATE: 
3.0 – 3-31-21 (revised); 4-1-21 (IRB approval) 
 
 
REVISION HISTORY 
 
Revision 
# 
Version 
Date 
Summary of Changes 
Consent 
Change? 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

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Table of Contents 
1.0 
Study Summary........................................................................................... 3 
2.0 
Objectives* .................................................................................................. 4 
3.0 
Background* ............................................................................................... 4 
4.0 
Study Endpoints*......................................................................................... 6 
5.0 
Study Intervention/Investigational Agent ..................................................... 6 
6.0 
Procedures Involved* .................................................................................. 7 
7.0 
Data and Specimen Banking* ................................................................... 12 
8.0 
Sharing of Results with Subjects* ............................................................. 13 
9.0 
Study Timelines* ....................................................................................... 13 
10.0 Inclusion and Exclusion Criteria* ............................................................... 13 
11.0 Vulnerable Populations* ............................................................................ 14 
12.0 Local Number of Subjects ......................................................................... 14 
13.0 Recruitment Methods ................................................................................ 15 
14.0 Withdrawal of Subjects* ............................................................................ 15 
15.0 Risks to Subjects* ..................................................................................... 16 
16.0 Potential Benefits to Subjects* .................................................................. 16 
17.0 Data Management* and Confidentiality ..................................................... 16 
18.0 Provisions to Monitor the Data to Ensure the Safety of Subjects* ............. 17 
19.0 Provisions to Protect the Privacy Interests of Subjects ............................. 18 
20.0 Compensation for Research-Related Injury .............................................. 18 
21.0 Economic Burden to Subjects ................................................................... 18 
22.0 Consent Process ....................................................................................... 18 
23.0 Process to Document Consent in Writing .................................................. 20 
24.0 Setting ....................................................................................................... 20 
25.0 Resources Available ................................................................................. 20 
26.0 Multi-Site Research* ................................................................................. 21 
 
 
 

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1.0 
Study Summary 
 
Study Title 
Effects of Post-Exercise Recovery Drink Composition 
on Subsequent Performance in Masters Class Athletes  
 
Study Design 
Randomized, Double-blind, placebo-controlled 
between-subject  
Primary Objective 
The purpose of this study is to demonstrate that 
performance in a subsequent bout of high-intensity 
glycogen depleting exercise will not be significantly 
different regardless of post-exercise recovery drink 
(carbohydrate vs. carbohydrate and protein vs. 
placebo). 
Secondary 
Objective(s) 
Assess the level of sports nutrition knowledge and total 
daily carbohydrate and protein intake in a group of 
endurance-trained masters class athletes.  
Research 
Intervention(s)/ 
Investigational 
Agent(s)  
Erica Goldstein, MS, RDN 
Jeffrey Stout, PhD  
David Fukuda, PhD  
Adam Wells, PhD 
Maxine Furtado, BS  
IND/IDE #  
 
Study Population 
Male Athletes, Age 35-59 years  
Sample Size 
50 
Study Duration for 
individual 
participants 
14 days; 3-4 total visits  
Study Specific 
Abbreviations/ 
Definitions  
Graded Exercise Test (GXT), Maximal Oxygen 
Consumption (VO2max), Plateau in Oxygen Uptake 
(VO2peak), High-Intensity Aerobic Interval Exercise 
(INT), Time to Exhaustion (TTE), Heart Rate Variability 
(HRV), Sweat Rate (SR), Carbohydrate (CHO), 
Carbohydrate-Protein (CHO-P), Placebo (PLA), 
Masters Class Athletes (MCA’s) 
 
 

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2.0 
Objectives* 
2.1 The purpose of this investigation is to examine the effects of 
carbohydrate vs. carbohydrate-protein co-ingestion vs. 
placebo on a subsequent bout of performance following high-
intensity exercise and a 2-hour recovery period in Masters 
Class Athletes. 
2.2 It is hypothesized that: 
• Carbohydrate will improve subsequent performance 
significantly better than the placebo post-exercise in 
male Masters Class Athletes. 
• Carbohydrate and protein will improve subsequent 
performance significantly better than PLA post-
exercise in male Masters Class Athletes.   
• Carbohydrate and Protein will not improve 
subsequent performance significantly better than 
CHO post-exercise in male Masters Class Athletes. 
3.0 
Background* 
3.1 The rate of participation by masters class athletes (MCA’s) in 
organized endurance and ultra-endurance (>6 hours) events 
has increased exponentially over the past 10-15 years (Lepers 
et al. 2013; Lepers and Stapley 2016). MCA’s are generally 
classified as ≥35 years of age who meticulously train for and 
compete in organized sport (Ransdell et al., 2009; Lepers and 
Cattagni 2012; Lepers and Stapley 2016; Fien et al. 2017). 
Carbohydrate and protein are key nutritional factors that 
promote glycogen and protein synthesis, support recovery and 
allow for repetitive high-intensity efforts. Few studies, 
however, have examined short-term (2-6 hours) post-exercise 
recovery nutrition specific to MCA’s (Doering et al. 2016a, 
2017).  
MCA’s (cyclists) maintain weekly mileage of approximately 
250 – 350 miles per week, with no significant difference 
between younger (35-44 years) and older (≥ 55) age groups. 
Endurance exercise at this level of competition requires 
prolonged high-intensity activity of ~70-90% VO2max, which 
relies quite heavily on carbohydrate oxidation (Romijn et al. 
1993; Spriet 2007). Competitive athletes rely on their ability to 
both store and mobilize glucose from muscle. The storage 
form of glucose in muscle is glycogen. Typical glycogen 
depleting protocols in studies that have examined the 
difference between carbohydrate (CHO) ingestion or 
carbohydrate-protein (CHO-P) co-ingestion have been running 

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or cycling to fatigue at 70% VO2max or intermittent exercise at 
70%-90% VO2max. These protocols have been sufficient for 
depleting muscle glycogen content to ~100 – 200 mmol/kg/dw 
(Tsintzas et al. 1995; Jentjens et al. 2001; Alghannam et al. 
2016; Dahl et al. 2020). Conversely, both CHO (1.2 g/kg) and 
CHO-P co-ingestion (0.8/0.4 g/kg) were found to be equally 
effective for promoting glycogen resynthesis during varying 
periods of acute recovery (3, 4, and 5 hours) (van Loon et al. 
2000; Jentjens et al. 2001; Alghannam et al. 2016; Dahl et al. 
2020). 
Consuming carbohydrates immediately post-exercise has 
been shown to increase glycogen resynthesis rate by three 
times as compared to delaying carbohydrates for 2-hours (Ivy 
et al. 1988). Ivy et al. (1988) demonstrated the rate of 
glycogen synthesis to be 45% slower when delaying 
carbohydrate intake by 2 hours. Consequently, muscle 
glycogen concentration was also significantly less when 
carbohydrate intake was delayed by 2 hours following high-
intensity intermittent exercise (Ivy et al. 1988). A similar 
pattern was shown for leg protein synthesis when a CHO-P 
supplement (10 g protein; 8 g carbohydrate; 3 g fat) was 
provided immediately post-exercise in comparison to 3 hours 
later (Levenhagen et al. 2001). 
Two different isoenergetic options have been shown to 
effectively promote glycogen resynthesis post-exercise: CHO 
only (1.2 g/kg) or CHO-P co-ingestion (0.8 g/kg/0.4 g/kg). 
Several studies had demonstrated an equivalent increase in 
muscle glycogen content (van Loon et al. 2000; Jentjens et al. 
2001; Alghannam et al. 2016) when these recovery beverages 
were consumed beginning immediately post-exercise and at 
30-min intervals after that for 3, 4, and 5-hour recovery 
periods, or an increase in favor of CHO-P co-ingestion when 
consumed immediately-post and at 1-hour intervals during the 
first 2 hours (Berardi et al. 2006).  
MCA’s may train twice per day, with limited time between 
bouts of exercise; therefore, an effective recovery strategy is 
paramount. On two occasions, CHO-P co-ingestion has been 
associated with either increased performance or an attenuated 
decrement to performance during a subsequent bout of 
endurance exercise in younger athletes (Berardi et al. 2008; 
Dahl et al. 2020). Appropriate post-exercise nutrient intake is 
vital for MCA’s as several studies have reported slower 
recovery from muscle-damaging exercise, even when 
moderate amounts of protein are consumed (Doering et al. 
2016a, 2017).  

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Post-exercise nutrient intake in a group of Australian 
(endurance) MCA’s was reported to be well below the 
suggested amount for CHO (~0.7 g/kg) and slightly below the 
recommended threshold for protein (~0.3 g/kg) (Doering et al. 
2016b). Furthermore, the MCA’s surveyed in this study 
demonstrated poor or no knowledge of post-exercise 
nutritional practices for CHO and protein intake (Doering et al. 
2016b). It has been suggested that while endurance athletes 
are familiar with the need for carbohydrates in the diet, the 
importance of meeting total daily protein needs as well as the 
incorporation of protein within the immediate-post recovery 
period may be less well understood (Vitale and Getzin 2019). 
Inadequate nutrient intake in the acute post-exercise recovery 
period may compromise the rate of both glycogen and protein 
synthesis. On a cellular level, this may negatively affect 
skeletal muscle remodeling and glycogen storage, promoting 
fatigue and poor training outcomes and performance (Ivy et al. 
1988; Breen et al. 2011; Kruseman et al. 2020).  
In conclusion, the rate of MCA participation in elite-level 
endurance competition is exponentially increasing. In contrast, 
the level of nutritional knowledge and recovery practices was 
reportedly poor in one group of surveyed endurance MCA’s 
(endurance) athletes. Additionally, a critical gap in post-
exercise recovery research exists specific to MCA’s. Even less 
research exists specific to the MCA and short-term (2 hours or 
less) recovery. This is a problem as slower recovery rates 
between subsequent bouts of exercise have been 
demonstrated, and higher protein needs post-exercise have 
been indicated to promote muscle remodeling.  A unique 
opportunity exists to examine CHO-P co-ingestion in a 2-hour 
recovery period to promote optimization of post-exercise 
glycogen resynthesis and protein remodeling to support 
successive bouts of training or competition. 
 
4.0 
Study Endpoints*  
4.1 The study will be completed when participants have finished 
all visits to the laboratory. It is anticipated that the total testing 
for each individual should be completed within 14 days or 3-4 
total visits.  
5.0 
Study Intervention/Investigational Agent 
1.1 Participants will consume either a CHO beverage (1.2 g/kg of 
bw), a CHO-P beverage (0.8 g/kg bw CHO + 0.4 g/kg bw 
protein), or PLA beverage matched for taste and appearance 
between exercise bouts to elucidate which drink moderates 

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performance decrements most effectively. The dietary source 
of CHO will be Gatorade®, the PLA will be Gatorade® G Zero, 
and the protein will be provided via BiPro Elite whey protein 
isolate.  
 
6.0 
Procedures Involved* 
6.1 To test the hypothesis, this study will utilize a randomized, 
double-blind, placebo-controlled between-subject design to 
examine the effects of CHO and CHO-P supplementation on 
short-term recovery following aerobic interval exercise and 
time to exhaustion testing in MCA’s. After completing the 
informed consent and screening process (Visit #1), each 
participant will be assessed for their anthropometric measures 
as well as body composition and will complete a GXT to 
volitional exhaustion on a cycle ergometer to determine 
VO2peak. Participants will also be provided with the option to 
return to the lab on a separate date for the GXT and VO2peak 
testing. Forty-eight to 72 hours after Visit #1, or VO2peak 
testing, participants will return to the lab for familiarization with 
the INT exercise protocol and TTE (Visit #2). Within five to 
nine days after familiarization (Visit #2), participants will return 
to the lab for Visit #3 to complete INT exercise and TTE, a 2-
hour passive recovery period including the consumption of one 
of three treatments (CHO, CHO-P, PLA), followed by repeated 
INT exercise and TTE, HRV and SR determination. 
Participants will also complete the Nutritional Recovery 
Practices, Knowledge, and Beliefs of Australian Triathletes 
survey during the 2-hour recovery period. Participants will be 
asked to complete the ASA24®, an automated self-
administered 24-hour dietary assessment tool during the 5-9-
day period between familiarization (Visit #2) and the 
experimental protocol (Visit #3). 
Testing Procedures.  
• Participants will report to the testing location on 3 
separate occasions.  
• Visit #1: The study will be explained to the participant 
by the investigators. Participants will be asked to 
provide their written, informed consent. The 
participants will then be asked to complete the 
American College of Sports Medicine Exercise 
Preparticipation Health Screening Questionnaire for 
Exercise Professionals (PHSQEP), Physical Activity 
Readiness Questionnaire for Everyone (PAR-Q+), 
and Medical Health and Activity Questionnaire. 

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Following the written consent and screening process, 
anthropometrics and body composition will be 
measured, and the VO2peak test outlined in section 
6.3 will be completed. Following informed consent, 
screening, and anthropometrics and body 
composition, if necessary (i.e., due to time 
constraints), participants may choose to return to the 
lab on a separate occasion for VO2peak testing.  
• Visit #2: Participants will report to the testing location 
for familiarization to the testing protocols. Based on 
VO2peak data obtained during Visit #1, participants 
will perform the INT and TTE protocol outlined in 
section 6.4 to familiarize them with the protocol for the 
experimental trial (Visit #3), and teach them how to 
respond to the rating of perceived exertion RPE 
measures.  
• Visit #3: To ensure participants are hydrated prior to 
the start of testing, they will be asked to consume 500 
ml of water 2 hours before their arrival to the 
laboratory for testing (Moreno et al. 2013; Vanderlei et 
al. 2015). Participants will complete a warm-up 
followed by the INT and TTE protocol, as outlined in 
section 6.4. Immediately following the INT and TTE 
testing, the participant will be given an isocaloric CHO 
(1.2 g/kg of bw), CHO-P (0.8 g/kg bw CHO + 0.4 g/kg 
bw protein), or PLA beverage that have a similar 
appearance and taste and will be blinded to the 
contents. Participants will rest for two hours and then 
repeat the INT and TTE testing (6.4). Participants will 
be weighed in their cycling bib shorts immediately 
before and after the first and second bouts of INT 
exercise and TTE testing. Sweat rate will be 
determined based on the difference between pre-and 
post-weight changes for each bout of exercise, the 
amount of fluid consumed during the recovery period, 
and total exercise time in minutes (Casa 2019). 
Participants will also be asked to report total sleep in 
number of hours and minutes for the night prior to 
testing. Immediately upon completion of the second 
exercise bout, participants will rest for an additional 
30 minutes for HRV recording. Completion of Visit #3 
will conclude all required testing.  
6.2 Anthropometric measurements: 
• Participants will be assessed for body weight, height, 
and body composition.  

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• Height will be assessed using a stadiometer (Health-
o-meter Professional Patient Weighing Scale, Model 
500 KL, Pelstar, Alsip, IL, USA), and body 
composition and weight will be assessed using Bio-
electrical Impedance Analysis (InBody 770, Biospace 
Co, Ltd. Seoul, Korea). Participants will be asked to 
remove any jewelry, their footwear, including socks, 
and wear only light athletic attire. Then they will be 
asked to stand on a platform while holding two 
handles out to the side. They will hold this position as 
the bioelectrical impedance analysis sends a minute 
electrical current (that is safe and cannot be seen or 
felt) through the body, to determine body composition.  
There are no risks or discomforts associated with the 
use of bioelectrical impedance analysis. Participants 
should be two-hours fasted for this assessment. 
6.3 Determination of VO2peak 
• All participants will perform a graded exercise test 
(GXT) to volitional exhaustion on a cycle ergometer 
(Lode, Corival cpet, Groningen, The Netherlands) to 
determine VO2peak and peak power output. Prior to 
testing, each participant will be provided with a Polar 
heart rate monitor (chest strap and sensor; Polar H10, 
Polar Electro Oy, Kempele, Finland) to record heart 
rate.  Participants will be asked to place the heart rate 
monitor just below the sternum. Participants will 
complete a five-minute warm-up on the cycle 
ergometer at a self-selected intensity and cadence. 
The test will consist of 2-minute stages, beginning at 
an initial workload of 50 watts (W), then 100W, then 
150W followed by an increases of 30W every 2 
minutes until the participants can no longer maintain 
60 revolutions per minute (rpm) (Beltz et al. 2016; 
McCarthy and Spriet 2019). VO2peak will be 
determined as the highest peak value achieved during 
the last completed stage of the test, and if it coincides 
with at least two of the following three parameters: 
heart rate (HR) within 10% of age-predicted maximal 
HR; respiratory exchange ratio (RER) of 1.15 or 
higher; a plateau in oxygen consumption despite an 
increase in exercise intensity. Open-circuit spirometry 
will be used to estimate VO2peak (l·min-1) with a 
calibrated metabolic cart (True One 2400® Metabolic 
Measurement System, Parvo-Medics Inc., Sandy, UT) 
by sampling and analyzing the breath-by-breath 

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expired gases. The metabolic cart software 
continuously records ventilation and expired gases, 
with averages every 15s, calculates VO2, and 
determines the VO2peak value. The highest power 
output achieved will be recorded as peak power 
output (PPO) in watts. 
6.4 High-Intensity Aerobic Intervals and Time to Exhaustion 
• All participants will warm-up on a cycle ergometer for 
3-minutes at 50W, 2-minutes at 100W, and 1-minute 
at 75W followed by a 3-minute rest. Participants will 
then perform 5 x 4-minute high-intensity aerobic 
intervals at 70-80% of individual PPO, as determined 
by the familiarization trial. Two-minutes of low-
intensity cycling at 50W will separate intervals. 
Immediately following the fifth interval, participants will 
cycle at a work rate (watts) that corresponds to 90% 
PPO until volitional exhaustion or when rpm falls 
below 60 for 10 seconds. Following the endurance 
trial (INT and TTE), participants will cycle with no 
resistance for 5 minutes to cool down. Revolutions 
per minute will be the only performance measure 
visible to participants, and minimal verbal 
encouragement will be provided throughout the 
aerobic intervals and time to exhaustion. Rating of 
perceived exertion (RPE) will be recorded using the 
Borg 10 scale during the last 30 seconds of each 
aerobic and rest interval and once immediately before 
completion of the TTE. 
6.5 Recovery Slope of Heart Rate Variability 
• R-R intervals recorded via the heart rate monitor and 
Elite HRV will be downloaded and analyzed using a 
laptop with commercially available HRV analysis 
software (Kempele, Finland; Kubios HRV Analysis v 
3.3, Kuopio, Finland). R–R interval series will be 
filtered using the software’s automatic artifact 
correction to detect and replace artifacts from the 
normal sinus arrhythmia. Replaced R–R periods will 
not exceed 5% of total heartbeats. Each window will 
be analyzed for the time-domain values of root mean 
square of the successive differences of R–R intervals 
(RMSSD). RMSSD reactivation following exercise will 
be reported to behave linearly during the first 30-
minutes post-exercise (Orellana et al. 2019). 
Therefore, RMSSD values will be recorded during the 
90% TTE and then post-exercise every 5 min.  Using 

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simple linear regression, a slope value for RMSSD 
values vs. time of recording (end of TTE, 5min, 
10min, 15min, 20min, 25min, 30min) for each 
participant will be performed. 
6.6 Dietary Analysis 
• To understand if MCA are meeting carbohydrate and 
protein intake recommendations, participants will be 
asked to complete the ASA24®. The ASA24® is a 
validated, automated self-administered 24-hour 
dietary assessment tool developed by the National 
Cancer Institute (Bethesda, Maryland) (Park et al. 
2018). Each participant will be instructed on the use 
of the ASA24® during Visit #1, either by a registered 
dietitian or a trained volunteer research assistant. 
Participants will be asked to report a typical daily 
intake for days that they train. Following completion of 
VO2peak testing participants will be emailed a sample 
of a detailed diet recall and an individual username 
and password to access the ASA24® system. They 
will be asked to report a 24-hour intake that is typical 
for days that they train. The sample diet recall will 
demonstrate a level of detail that includes all foods 
and fluids consumed upon waking until bedtime, 
including foods and/or supplements consumed pre, 
during, and immediately post-exercise. Participants 
will be asked to complete the electronic diet recall 
during the 5-9-day period between familiarization 
(Visit #2) and the experimental protocol (Visit #3). 
After data collection, the dietary intake will also be 
analyzed for total energy (kilocalorie [kcal]) and 
macronutrient distribution (carbohydrate, protein, and 
fat). An Excel spreadsheet of each participant’s 
macro-and micronutrient analysis will be downloaded 
from the ASA24® website. 
6.7  Postexercise Nutritional Practices Survey  
• The Nutritional Recovery Practices, Knowledge and 
Beliefs of Australian Triathletes survey is an 
instrument previously designed to assess knowledge 
of postexercise nutritional recommendations and 
recovery practices in endurance athletes (Doering et 
al. 2016b). Participants will complete the survey 
electronically during the 2-hour recovery period (Visit 
#3).  Masters class athletes in this study will be asked 
to complete a series of 34 total questions assessing 
their postexercise nutrition knowledge and recovery 

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practices. Questions 1 through 10 will be omitted as 
the current study is not specific to triathletes. 
6.8 
Outcome Variables  
• VO2peak Test 
o VO2peak 
o Peak Power  
• INT and TTE Protocol  
o Peak Power  
o Average Power  
o RPE  
o Total time to exhaustion at 90% VO2peak 
• Anthropometrics  
o Height  
o Weight 
o Age  
o Body Fat Percentage  
• HRV  
o Root mean square of the successive 
differences of R–R intervals (RMSSD) 
• Sweat Rate Determination  
o Individual sweat rate pre-post exercise  
7.0 
Data and Specimen Banking* 
7.1 
Data Storage.  
All informed consents, medical history questionnaires and 
screening paperwork will be stored in a locked cabinet in the 
Physiology of Work and Exercise Response (POWER) 
Laboratory, and data collection sheets will be stored in a 
locked cabinet in the Kinesiology and Teaching Laboratory 
during and following the investigation; all electronically 
entered data will be saved in an encrypted file. Data from 
VO2peak tests and performance tests will be collected and 
managed using Trueone 2400 software on a secure 
computer. The computer is password-protected, and the 
data stored is deidentified and password protected. 
Participant folders will be marked with an ID number to    
protect against a breach of confidentiality; participant names 

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and ID numbers will be stored separately in a digitally 
encrypted file. 
7.2    Data Access. 
Access to research-related data, paperwork, and records will 
be limited to appropriate laboratory personnel only. 
7.3 
Data disposal. 
 
All participant records and paperwork (e.g., informed consent 
forms, data collection sheets) will be destroyed six years from the 
end of the study. Password protected electronic file containing 
participants’ names will be discarded after the completion of data 
analysis. 
 
7.4 
Data dissemination 
The results of this study will be published as a group as part 
of a scientific publication. No individual results will be 
published or shared with any third person or party. 
8.0 
Sharing of Results with Subjects* 
8.1 Individual results will remain confidential and will only be 
relayed to the participant upon request. All information 
attained from the medical history questionnaire will be held in 
strict confidence. 
9.0 
Study Timelines* 
9.1 Describe: 
• All visits will be separated by 48-72 hours. Visit #1 
and Visit #2 will take approximately 60-90 min each. 
Visit #3 will take approximately 4 hours. 
• We anticipate the participant recruitment and 
enrollment will be completed within two months and 
estimate that data collection will be completed within 
approximately 6 to 8 months. 
• The estimated date for completion of primary analysis 
will be December 2021. 
 
10.0 Inclusion and Exclusion Criteria* 
10.1 Individuals will be screened for eligibility using the American 
College of Sports Medicine Exercise Preparticipation Health Screening 
Questionnaire for Exercise Professionals (PHSQEP), Physical Activity 
Readiness Questionnaire for Everyone (PAR-Q+), and confidential 
Medical Health and Activity Questionnaire to determine the need for 
medical clearance from a health care professional. Participants presenting 

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any sign or symptoms suggestive of cardiovascular, metabolic, or renal 
disease will be asked to seek medical clearance in order to participate. 
10.2 Inclusion Criteria. 
• Men between the ages of 35 and 59 years 
• Free of any physical limitations as determined by the 
Exercise Preparticipation Health Screening 
Questionnaire for Exercise Professionals (PHSQEP), 
and a Physical Activity Readiness Questionnaire for 
Everyone (PAR-Q+) 
• Regularly engaged in endurance exercise (running, 
cycling, swimming) for a minimum of three years, a 
weekly training volume of 5-10 hours, and a maximal 
oxygen uptake of 45.00 ml·kg-1·min-1 or higher as 
determined by completion of VO2peak assessment 
(Visit #1). 
10.3 Exclusion Criteria 
• Recent musculoskeletal injuries or surgeries  
• Any chronic illness that requires continuous medical 
care 
• Current incarceration, cognitive impairment or inability 
to provide consent 
• Present or past use of performance-enhancing drugs 
10.4 Indicate specifically whether you will include or exclude each 
of the following special populations:  
• 
Adults unable to consent: exclude 
• 
Individuals who are not yet adults (infants, children, 
teenagers): exclude  
• 
Pregnant women: exclude 
• 
Prisoners: exclude 
11.0 Vulnerable Populations* 
11.1 N/A: This research does not involve any vulnerable populations. 
• 
No pregnant women will be included. 
• 
No prisoners will be included. 
• 
No children will be included. 
• 
No cognitively Impaired Adults will be included. 
12.0 Local Number of Subjects 
12.1 All participants will be recruited from the University of Central 
Florida and from within the surrounding Central Florida area. 
 

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12.2 Participants will be recruited and screened until a maximum of 50 
individuals are enrolled. Participants will be considered enrolled only after 
an informed consent document has been signed, a medical history 
questionnaire, PHSQEP and PAR-Q+ have been completed, and 
anthropometric data have been recorded. To account for individuals who 
do not meet the inclusion criteria, the number of recruited individuals will 
likely exceed the number of enrolled participants. However, no more than 
50 participants will be recruited. 
 
 
13.0 Recruitment Methods 
13.1 Participants will be recruited at the University of Central Florida and 
in surrounding locations through flyers, posters, information sheets, 
notices, and internet postings and media to recruit participants. The flyer 
will be posted on social media. Participant recruitment will begin upon 
approval of the IRB protocol and all supporting documents. 
 
13.2 Subjects will primarily be recruited from the University of Central 
Florida and surrounding locations. Participants will be eligible to 
participate if they hear about the study through flyers, posters, information 
sheets, notices, internet postings, and/or media, and word of mouth.  
 
13.3 Participants will be recruited by flyers, posters, information sheets, 
notices, and internet postings, which will detail the study objectives and a 
brief explanation of the inclusion and exclusion criteria. 
 
13.4 There will not be any compensation to participants for participating 
in this study 
 
14.0 Withdrawal of Subjects* 
14.1 Participants have the right to discontinue participation without 
penalty, regardless of their status in the study. Participants will 
also be instructed that participation in the study may be 
terminated at any time by the researchers in charge of the 
project. Possible reasons for subject withdrawal without their 
consent include inability or unwillingness to adhere to the 
study protocol, failure to adhere to any requirements (including 
exercising vigorously within the 24-hours prior to the testing 
visit), and refusal to participate in testing trials 
14.2 If a participant is asked to withdraw without their consent, they 
will be told that they can no longer participate in the study and 
the reasoning for this will be explained privately. Additionally, 
study personnel will explain the destruction of their data to 
them. 

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14.3 If a participant is asked to withdraw or voluntarily withdraws 
from the study, their data will be removed from the study 
results and destroyed. 
15.0 Risks to Subjects*  
15.1 The exercise-based assessments carry the same inherent 
risks as participating in any physical activity, such as muscle 
soreness, and fatigue and possibly muscle strains, and/or joint 
sprains, elevated heart rate, feelings of nausea and possible 
episodes of emesis, arrhythmia, myocardial infarction, and 
death. To minimize these risks, participants will be instructed 
on the appropriate technique for the performance 
assessments and will be required to complete a warm-up prior 
to completing the assessments. Furthermore, participants will 
be informed that they can quit at any time before or during the 
test. All personnel involved in this study are CPR certified. 
Furthermore, all research personnel involved in data collection 
are experienced in the administration of the proposed 
assessments, and all Doctoral students involved in this study 
are Certified Strength and Conditioning Specialists through the 
National Strength and Conditioning Association. Participants 
will be instructed to immediately stop and report any injury or 
discomfort associated with the performance assessments to a 
member of the investigative team. The extent of the 
injury/discomfort, as well as the ability of the participant to 
continue with the study, will be subsequently be determined by 
the investigative team. If it is deemed that the discomfort/injury 
will prevent the participant from completing the study, or if the 
injury/discomfort may be exacerbated by further participation 
in the study, the investigative team will suspend the 
individual’s participation in the study. 
15.2 This study is recruiting for male participants only. Therefore, 
subjects who are pregnant will not be enrolled.  
15.3 There are no risks to others in the study who are not 
participants. 
16.0 Potential Benefits to Subjects* 
16.1 There is no direct benefit of participation in the study. 
17.0 Data Management* and Confidentiality 
17.1 All statistical analyses will be conducted via the Statistical 
Package for Social Science (SPSS) software for Windows version 
21 (SPSS Inc., Chicago, IL). Data will be statistically analyzed 
using separate 1-way analysis of covariance (ANCOVA) for TTE, 
HRV-Slope Recovery, and Sweat Rate measures. The pretest and 

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the posttest values will be used as a covariate and dependent 
variable, respectively. If the ANCOVA is significant, Fisher’s LSD 
Post-Hoc analysis will be used to determine group differences. For 
effect size (ES), the partial eta squared statistic will be reported, 
with 0.01, 0.06, and 0.14 representing small, medium, and large 
ES, respectively. The significance level will be set at p ≤ 0.05. 
17.2 The calculation of sample size (G*Power 3.1.9.4) was based 
on pilot data in low-to-moderately trained young men (22.1 + 
3.2 years) collected in our lab using the same protocol (90% 
PPO TTE) as outlined in this study. The total sample size was 
determined to be a total of 18 (6 in each group) based on a 
one-way ANCOVA, power of 0.90, and an alpha of 0.05. 
However, due to potential dropout rates, a total of 30 
participants (10 per group) will be recruited with the goal of 
completing 18. 
17.3 Individual results will remain confidential and only be related to 
the participant upon request once the study is completed. All 
questionnaires and data collection sheets will be kept in a 
locked cabinet during and following the study. Data that is 
electronically stored will be kept on a password-protected 
computer in the office of the investigator who oversees data 
collection. All information will be destroyed six years from the 
end of the study. Participant folders will be marked with an I.D. 
number to protect against a breach of confidentiality. 
Participant names and I.D. numbers will be stored apart from 
the subject folders in a password protected electronic file. 
17.4 Quality control of collected data will be performed by ensuring 
all data files remain in filing cabinets when not in use. 
17.5 Participant folders and other data will be kept in a locked filing 
cabinet and stored on a computer whose files are encrypted with a 
password. Further, all identifiable information will be stored separately 
from any data collection sheets; Computer file containing participants’ 
names will be discarded upon completion of data analysis. Only study 
personnel will have access to data. All signed consent forms will be stored 
separately from the rest of the data and will be held for a minimum of 5 
years after completion of the study. 
 
18.0 Provisions to Monitor the Data to Ensure the Safety of 
Subjects*  
This section is required only when research involves more 
than Minimal Risk to subjects 
18.1 Participants will be instructed to immediately stop and report 
any injury, discomfort, or concern associated with the 

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performance assessments. Any new information or adverse 
event will be reported to the IRB. Safety data will be reviewed 
by the study investigators at the time of any reported incidents. 
The investigator in charge of data collection will consult with 
the principal investigator in case of injury or concern raised by 
the participants. 
19.0 Provisions to Protect the Privacy Interests of Subjects 
19.1 Participant privacy will be protected at all times. All data 
collection processing pertaining to this study will be solely 
conducted by its own personnel. No other persons will have 
access to the previously mentioned facilities during data 
collection and processing. Participants will be provided locker 
room access to change into their attire if needed. 
19.2 All consent forms will be completed privately in a small office 
at the University of Central Florida. All performance 
assessments and testing sessions will be conducted in the 
Kinesiology Laboratories. Only the primary investigators on 
this study will have access to any personal information which 
will be stored privately in a locked file cabinet. All assessments 
will take place in a laboratory that is not accessible to the 
general public. 
19.3 Only the approved principal investigator and co-investigators 
will be permitted to access any sources of information about 
the subjects, which will be in locked cabinets and password-
protected computers and files. 
20.0 Compensation for Research-Related Injury 
20.1 Participants are instructed to report any discomforts related to 
the study to the principal investigator. If immediate assistance 
is needed, it will be provided via the emergency medical 
system. For non-emergency discomforts, participants must 
seek their own physician for medical attention. Adverse 
events/side effects will be reported to the IRB immediately 
upon notification. 
21.0 Economic Burden to Subjects 
21.1 Participants will be responsible for providing their own means 
of transportation and parking to and from the Kinesiology 
Laboratories at the University of Central Florida for testing 
sessions. 
22.0 Consent Process 
This study will require a written form of consent for subject 
participation  

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• 
The consent process will take place in a private room at 
the University of Central Florida. 
• 
All participants will be provided with the time necessary 
to read all documents, and an investigator will be 
available to explain the study protocol and answer any 
questions that each potential participant may have. 
• 
Ongoing consent will be obtained by verbal agreement of 
the participant to come in for future testing sessions. 
• 
This study will utilize “SOP: Informed Consent Process 
for Research (HRP-090)”. 
 
Include if there are Non-English Speaking Subjects, 
otherwise delete. 
• 
N/A: Participants who only speak languages other than 
English will not be enrolled.  
Include if you are requesting Waiver or Alteration of 
Consent Process (consent will not be obtained, required 
information will not be disclosed, or the research involves 
deception), otherwise delete. 
• 
N/A: A waiver or alteration of consent process will not be 
used 
Include if you are enrolling subjects who are not yet 
adults (infants, children, teenagers), otherwise delete. 
• N/A: Subjects who have not attained the legal age for 
consent to treatments or procedures involved in research 
will not be enrolled.   
Include if you are enrolling Cognitively Impaired Adults, 
otherwise delete. 
• 
N/A: Adults who are cognitively-impaired will not be 
enrolled. 
Include if you are enrolling Adults Unable to Consent, 
otherwise delete  
• 
N/A: Adults unable to consent will not be enrolled.  
 

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23.0 Process to Document Consent in Writing 
23.1 This study will be following the guidelines of “SOP: Written 
Documentation of Consent (HRP-091). 
23.2 This research requires written documentation of consent. 
23.3 Consent form attached using “CHECKLIST: Waiver of 
Written Documentation of Consent (HRP-411).” 
 
24.0 Setting 
24.1 All testing procedures will take place at the University of 
Central Florida in the Physiology of Work & Exercise 
Response Laboratory in the Education Complex. 
• 
The research team will identify and recruit potential 
subjects at the University of Central Florida and in 
surrounding locations through Advertisements, flyers, 
posters, notices, internet postings, and/or media. 
• 
All research procedures will be performed at the 
University of Central Florida in the Education Complex. 
• 
There will not be any involvement of a community 
advisory board. 
• 
No research will be conducted outside of the University of 
Central Florida. 
25.0 Resources Available 
25.1 All research personnel are currently certified in adult 
cardiopulmonary resuscitation (CPR) and automated external defibrillator 
(AED) use. An AED is located outside of the Kinesiology Laboratories 
within the Education Building. All the assessments will be monitored by a 
Certified Strength and Conditioning Specialist (National Strength and 
Conditioning Association).  
 
25.2 Our research team is experienced and has strong expertise in 
similar types of research projects. As collaborating researchers in UCF’s 
School of Kinesiology and Physical Therapy, our team has published 
dozens of peer-reviewed research articles covering areas of nutritional 
supplementation, exercise, and physical performance involving subjects of 
varying ages, training levels, and health statuses. 
 
25.3 The Kinesiology laboratories are research spaces featuring some of 
the latest technologies and instrumentation to evaluate human 
performance on many levels, ranging from physical functioning of the 
whole body to molecular and signaling pathways underlying such 
functioning. Beyond our exercise equipment to assess muscular strength 
and endurance, our facility is also equipped with instruments designed to 
assess body composition and non-invasive, in vivo imaging of the 

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musculoskeletal system and vasculature. Additionally, our laboratory 
features devices used to evaluate the visual-motor function and motor 
skills, electrical activity of muscles, and energy expenditure. This facility is 
conducive to exercise training and supervision. 
 
25.4 Prior to data collection and participant recruitment, all personnel 
involved in the investigation will be provided with copies of all study 
materials for review. All personnel will meet to discuss the study protocol, 
research procedures, and individual duties. Additionally, all persons 
assisting with the research will be appropriately trained in laboratory safety 
and the treatment of human subjects. Besides interacting with human 
subjects, data analysis and preparation will occur on equipment in one of 
the Kinesiology Laboratories and the study investigators’ secured offices. 
 
 
26.0 Multi-Site Research* 
N/A. 
References 
 
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Beltz NM, Gibson AL, Janot JM, et al (2016) Graded Exercise Testing Protocols for the 
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J Sports Med (Hindawi Publ Corp) 2016:. https://doi.org/10.1155/2016/3968393 
 
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