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Page 1: The ergogenic effect of beta alanine on anaerobic endurance 13035885

The ergogenic effects of β-Alanine on anaerobic endurance

By

Ernest Didehvar-Sadr

Project Supervisor: Dr. Fang Lou

Key words: β-Alanine, carnosine, anaerobic, endurance, glycolysis, L-histidine

This investigation was conducted at the School of Life and Medical Sciences by Ernest Didehvar-Sadr in partial fulfilment of the requirements of the University of Hertfordshire for the degree of Bachelor of Science with Honours in Biomedical Science

School of Life and Medical SciencesUniversity of Hertfordshire Date: 22/6/16

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DECLARATION

I declare that:

(a) All the work described in this report has been carried out by me – and all the results (including any survey findings, etc.) given herein were first obtained by me – except where I may have given due acknowledgement to others;

(b) all the prose in this report has been written by me in my own words, except where I may have given due acknowledgement to others and used quotation marks, and except also for occasional brief phrases of no special significance which may be taken from other people’s work without such acknowledgement and use of quotation marks;

(c) all the figures and diagrams in this report have been devised and produced by me, except where I may have given due acknowledgement to others.

I understand that if I have not complied with the above statements, I may be deemed to have failed the project assessment, and/or I may have some other penalty imposed upon me by the Board of Examiners.

Signed …………………………… Date …………………………...

Name …………………………… Programme code ……………

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Acknowledgements

I wish to express my sincerest gratitude to my supervisor Dr. Fang Lou for her continuous

encouragement, backing and expert guidance throughout my research. I cannot thank her

enough for her patience and understanding through what has been the most challenging

endeavour throughout my time in academia, without her counsel and expertise, this

investigation would not have been a possibility.

I would also like to extend my deepest thanks to my father, Ahmad Didehvar-Sadr, MSc, who

read and critiqued my drafts, providing me with insight only years of experience could

provide, for that I am truly grateful. Finally, a special thanks goes to all the participants who

remained patient, stayed the course and followed the stringent protocol of the study, their

dedication and resilience served as an added source of inspiration for myself.

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ContentsACKNOWLEDGEMENTS...................................................................................................................... 3

LIST OF FIGURES:................................................................................................................................ 6

LIST OF TABLES:................................................................................................................................. 7

ABSTRACT:.......................................................................................................................................... 8

1. INTRODUCTION:........................................................................................................................... 9

1.1 Β-ALANINE: THE RATE LIMITING FACTOR.........................................................................................91.2 ADENOSINE TRIPHOSPHATE REGENERATION..................................................................................91.3 EXERCISE-INDUCED ACIDOSIS:.....................................................................................................91.4 CA2+ FUNCTION:......................................................................................................................... 111.5 ERGOGENIC POTENTIAL OF CARNOSINE:......................................................................................121.6 AIMS AND OBJECTIVE................................................................................................................. 131.7 HYPOTHESES............................................................................................................................. 13

2. MATERIALS AND METHODS:....................................................................................................14

2.1 SUBJECTS:................................................................................................................................ 142.2 SUPPLEMENTATION PROTOCOL...................................................................................................142.3 CROSSOVER DESIGN.................................................................................................................. 152.4 ANAEROBIC TESTING.................................................................................................................. 16

2.4.1 Parameters..........................................................................................................................162.4.2 Tempo................................................................................................................................. 162.4.3 Equipment........................................................................................................................... 172.4.4 Warming up......................................................................................................................... 17

2.5 STATISTICAL METHODS..............................................................................................................18

3. DATA AND RESULTS:................................................................................................................19

3.1 BENCH PRESS............................................................................................................................ 213.2 DEADLIFT.................................................................................................................................. 223.3 CHRONOLOGICAL ANALYSIS: THE LEARNING EFFECT....................................................................243.4 REPETITION VOLUME.................................................................................................................. 25

4. DISCUSSION:.............................................................................................................................. 29

4.1 ATP:CP TO ANAEROBIC GLYCOLYSIS.........................................................................................294.2 EXERCISE VOLUME AND ATP PRODUCTION..................................................................................304.3 MUSCLE COMPOSITION...............................................................................................................314.4 FUTURE CHALLENGES................................................................................................................. 32CONCLUSION:.................................................................................................................................... 34

5. REFERENCES:................................................................................................................................ 35

APPENDIX........................................................................................................................................... 37

1. ETHICS FORMS.......................................................................................................................... 37EC1: .......................................................................................................................................... 37EC2: .......................................................................................................................................... 58EC3: .......................................................................................................................................... 60EC6: .......................................................................................................................................... 61Risk assessment:.......................................................................................................................... 64

2. STATISTICAL TESTING................................................................................................................682.1: Bench Press:.........................................................................................................................682.2 Deadlift:.................................................................................................................................. 692.3: Deadlift chronologically ordered:............................................................................................702.4: Bench Press chronologically ordered:...................................................................................712.5: Deadlift 1st and 2nd quartiles:..................................................................................................722.6 Deadlift 3rd and 4th quartiles:...................................................................................................722.7 Bench press 1st and 2nd quartiles:...........................................................................................73

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2.8 Bench press 3rd and 4th quartiles:............................................................................................743. SAMPLE FOOD DIARY:................................................................................................................. 764. REPETITIONS COMPLETED..........................................................................................................77

4.1 Deadlift repetitions completed at each state of testing............................................................774.2 Bench press repetitions completed at each stage of testing...................................................784.3 Bench Press Time under tension at each stage of testing......................................................794.4 Deadlift time under tension at each stage of testing...............................................................80

5. HEALTH SCREEN TEMPLATE........................................................................................................81

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LIST OF FIGURES:

Figure 1.1: Substrates and products of the hexokinase reaction. Proton release originates from the hydroxyl group of the 6th carbon in glucose. Bond/group removal is represented by arrows facing away from a bond, addition of an atom or group is represented by arrows._____________________10Figure 1.2: Substrates and products of the phosphofructokinase (PFK) reaction. Proton release from comes against from the hydroxyl group of the 6th carbon, but this time of fructose 6-phosphate.____10Figure 1.3: Substrates and products of the glyceraldehyde 3-phosphate dehydrogenase reaction. Two electrons and a proton are used to form NADH from NAD+ following reduction. The remaining proton is released into solution (Robergs, Ghiasvand, & Parker, 2004)._____________________________10Figure 1.4: Graphical depiction of the regeneration of glycolytic ATP when coupled to ATP hydrolysis. This would occur during skeletal muscle contraction with no ATP contribution from mitochondrial respiration. The source of protons accumulating in the cytosol is ATP hydrolysis. The highlighted molecules are those left following the reactions (Robergs, Ghiasvand, & Parker, 2004).___________11Figure 2.1: A timeline describing the crossover design of the study in chronological order from subject participants acceptance into the study (left) to the completion of the final tests; test 2 (right)._______15Figure 2.2: (a) Depiction of hip circle in use to prepare for weightlifting by activating the muscles surrounding the synovial joint (sling shot hip circle, 2014). (b) A graphical representation of internal and external rotation of the rotator cuff using a dyna-band (Rotator cuff strain rehabilitation exercises, 2014). (c) The anatomy of a deadlift and the activated muscles in the posterior chain (Thomas, 2008). (d) The anatomy of a deadlift and the activated muscles in the anterior chain (Thomas, 2008)._____18Figure 3.1: Bar chart representing the mean bench press repetitions completed during baseline testing, post-placebo testing and post-β-alanine testing, with mean values labelled______________20Figure 3.2: Bar chart representing the mean deadlift repetitions completed during baseline testing, post-placebo testing and post-β-alanine testing, with mean values labelled.___________________20 Figure 3.3: Line chart displaying the changes in bench press repetitions completed between post-placebo testing and post-β-alanine testing for each participant, (mean difference ± SD= 2.68±2.23) 21 Figure 3.4: Bar chart depicting the mean difference in bench press repetitions completed between the 3 tests performed at the different stages of known supplementation. Mean differences displayed ± Standard error of the mean values.__________________________________________________22 Figure 3.5: Bar chart depicting the mean difference in deadlift repetitions completed between the 3 tests performed at the different stages of known supplementation. Mean differences displayed ± Standard error of the mean.________________________________________________________23 Figure 3.6: Line chart displaying the changes in deadlift repetitions completed between post-placebo testing and post-β-alanine testing for each participant, (mean difference ± SD= 2.86±1.73)______ 23 Figure 3.7: Mean deadlift repetitions completed during the consecutive stages of testing: baseline testing, test 1 and test 2 with mean values labelled._____________________________________24 Figure 3.8: Mean bench press repetitions completed during the consecutive stages of testing: baseline testing, test 1 and test 2, with mean values labelled. _____________________________25 Figure 4.1: Force records from the SR of rat skinned muscle fibres. Caffeine was applied to release SR Ca2+ producing the above contractures; thus the size of the contracture is an indication of the Ca2+ released in the SR. The middle recording follows the muscle be being exposed to 50 mm Pi for 20 s prior to washing off and caffeine application (Fryer, Owen, Lamb, & Stephenson).__________31

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LIST OF TABLES:

Table 1: Mean repetitions completed by each subject for the deadlift across the three stages of 2c vnsupplementation. 1st and 2nd quartile (mean repetitions±SD= 12.9±2.5) highlighted in blue, 3rd and 4th (23.1±6.3) quartile highlighted in green. Quartiles defined by by mean number of repetitions completed, with 1st and 2nd (lower 50%) having completed less than 3rd and 4th (greater 50%).___26Table 2: The mean repetitions completed by each subject for the bench press across the three stages of supplementation. 1st and 2nd quartile (mean repetitions±SD= 17.2±4.4) highlighted in blue, 3rd and 4th quartile highlighted in green (31.3±4.5)._____________________________________________27Table 3: Fiber types, carnosine content, and buffering capacity of the middle gluteal muscle of thoroughbred horse (Sewell, Harris, Marlin & Dunnett, 1992)._______________________________32

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Abstract:β-Alanine is a non-essential amino acid that has commercially been sold as a supplement to

increase muscle buffering capacity to regulate acidosis in high intensity anaerobic athletes.

The purpose of this study was to examine the effects of 10 days of β-Alanine

supplementation on muscular performance when performing endurance based strength

exercises. Twenty-two males (mean age 20.9±1.1 yrs) participated in a double-blind,

maltodextrin-controlled crossover study consisting of two randomly assigned groups.

Baseline testing followed by post-placebo and post-supplementation tests were performed on

participants which consisted of bench pressing (BP) (65% of bodyweight) for maximum

repetitions until failure, followed by deadlifting (DL) (120% of bodyweight) for maximum

repetitions until failure, the number of repetitions completed was recorded. A significant effect

(P<0.005) was found following one way ANOVA with repeated means testing (BP P=2.137E-

7***) (DL P=1.451E-8***), rejecting the H0. Post hoc, Bonferroni corrected pairwise tests

supported the alternative hypothesis (HA), finding a significant effect between both baseline

(BP P=3.200E-5***, mean difference= 2.773) (DL P= 4.509E-7***, mean difference= 2.773)

and post-placebo (BP P=4.100E-5***, mean difference= 2.681) (DL P= 3.851E-7***, mean

difference= 2.864) testing against post-β-Alanine testing, suggesting β-Alanine provides a

positive ergogenic effect on endurance.

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1. Introduction:

1.1 β-alanine: the rate limiting factor

3-aminopropanoic acid, commercially known as β-alanine, is a naturally occurring non-

essential amino acid that forms part of the dipeptides: carnosine, anserine and balenine. It’s

combination with L-histidine to synthesise carnosine in a bonding reaction regulated by the

enzyme carnosine synthase, provides the basis for its use a performance enhancing

supplement.

Discovered in 1900, along with it’s constituent amino acids β-alanine and histidine, by the

Russian scientists Gulewitsch and Amiradzibi, carnosine was found to have an imidazole ring

side chain with a pKa of 6.83 that in conjunction with it’s carboxyl and amino groups which

have pKa’s of 2.77 and 9.63 respectively, make it ideal for use as a pH buffer in physiological

conditions (Smith, 1938). This was originally theorised following the discovery of high

carnosine concentrations in species that were regularly exposed to bouts of hypoxia, such as

marine mammals, it was further confirmed when high histidine containing dipeptides were

found in large quantities in the muscles and brains of species that specialised in athletic

competition such as greyhounds, horses and humans (Sale, Saunders & Harris, 2009).

1.2 Adenosine Triphosphate regeneration

When exercise intensity surpasses steady state, there is a need for adenosine triphosphate

(ATP) regeneration from the phosphagen (ATP:CP) system and anaerobic glycolysis.

Following the rapid depletion of phosphocreatine supplying the ATP:CP system, anaerobic

glycolysis becomes the primary source of ATP. This form of glycolysis eventually results in

intramuscular acidosis which has been shown to be causative of fatigue-induced increases in

muscular activation and electromyographic amplitude (Maclaren et al., 1989), (Taylor, Bronks

& Bryant, 1997).

1.3 Exercise-induced acidosis:

The biochemistry of exercise induced acidosis has long been disputed and it was widely

believed that the dissociation of lactic acid as formed from pyruvate was directly causative of

the observed acidosis, hence the phenomenon was named lactic acidosis. However,

increasingly, evidence has been provided against this and suggested that H+ ion release due

to ATP hydrolysis in the hexokinase and phosphofructokinase reactions as well following the

oxidation of glyceraldehydayde-3-phosphate, were predominant causes of the decrease in

intramuscular pH (figure 1.1-1.3) (Robergs, Ghiasvand, & Parker, 2004) (Brooks,

Dubouchaud, Brown, Sicurello, & Butz, 1999.

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Figure 1.1: Substrates and products of the hexokinase reaction. Proton release originates from the hydroxyl group of the 6th carbon in glucose. Bond/group removal is represented by arrows facing away from a bond, addition of an atom or group is represented by arrows.

Figure 1.2: Substrates and products of the phosphofructokinase (PFK) reaction. Proton release from comes against from the hydroxyl group of the 6th carbon, but this time of fructose 6-phosphate.

Figure 1.3: Substrates and products of the glyceraldehyde 3-phosphate dehydrogenase reaction. Two electrons and a proton are used to form NADH from NAD+ following reduction. The remaining proton is released into solution (Robergs, Ghiasvand, & Parker, 2004).

Furthermore, it was argued that lactate production as part of the lactate dehydrogenase

reaction aids in supporting anaerobic glycolysis and also buffering against proton

accumulation within the muscle as cytosolic NAD+ is produced which supports continued

ATP regeneration via glyceraldehyde-3-phosphate oxidation during glycolysis (Robergs,

Ghiasvand, & Parker, 2004). Thus, the argument for H+ production during glycolysis being

causative of intramuscular acidosis hinges on the intensity of the exercise surpassing that of

steady-state. Beyond the point of steady-state, the level of ATP hydrolysis required to

provide the energy needed for the exercise surpasses the production capabilities of the

mitochondria, hence the H+ cannot be “removed” in the same fashion as they are in the

aerobic pathways (figure 2). Whether caused directly by lactate or not, lactate can still be

used as an accurate marker of cellular acidosis which inevitably occurs following anaerobic

glycolysis (Van Meerhaeghe & Velkeniers, 2005).

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Figure 1.4: Graphical depiction of the regeneration of glycolytic ATP when coupled to ATP hydrolysis. This would occur during skeletal muscle contraction with no ATP contribution from mitochondrial respiration. The source of protons accumulating in the cytosol is ATP hydrolysis. The highlighted molecules are those left following the reactions (Robergs, Ghiasvand, & Parker, 2004).

1.4 Ca2+ function:

Upon the occurrence of exercise-induced acidosis, fatigue will invariably follow, this occurs

due to inhibition of the excitation-contraction coupling. In summary, this is the complex

process in which an action potential triggers the release of Ca2+ from the sarcoplasmic

reticulum of the muscle, activating troponin C and stimulating myofibril contraction.

Acidosis has been theorised to negatively affect the coupling via three methods, the inhibition

of Ca2+ release, competitive inhibition of Ca2+ binding to troponin C and the limiting of actin

and myosin binding thus decreasing the force produced during the cross bridge cycle

(Stackhouse, Reisman, & Binder-Macleod, 2001). Of the three hypotheses, decreased pH

inhibiting the binding of Ca2+ to troponin C has been validated in study (Ball, Johnson, &

Solaro, 1994). Research into the inhibition of Ca2+ release was promising at first, showing

decreased secretion, however the same results were recreated without a decrease in pH

(Chin & Allen, 1998). As for decreasing the resultant force of the cross bridge cycle, the

original studies showing this were done below physiological temperatures, when repeated at

temperatures similar to those seen in vivo, this effect disappeared (Pate, Bhimani, Franks-

Skiba, & Cooke, 1995).

On top of acidosis’ impairing effect on the cross bridge cycle, inorganic phosphate (Pi)

accumulation following phosphagen system activation and also as a product of ATP

hydrolysis alongside adenosine 5'-diphosphate (ADP) has been implicated in muscle fatigue.

Pi is a more significant factor as it is produced both from the phosphagen system and

following the hydrolysis of ATP, it’s release from the myosin filament allows conformational

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change to occur while bound to actin thus providing the “power stroke” of muscle contraction

(McLester, 1997). However, an accumulation of Pi due to exercise metabolism can lead to

the reversal of this release and can therefore detrimental to force production of the muscle.

1.5 Ergogenic potential of carnosine:

From this, it could be surmised that if an individual were able to increase the time required for

acidosis to accumulate via buffering, that fatigue would take longer to affect the individual

when performing high intensity exercises, particularly in which anaerobic sources of energy

are required such as glycolysis. This was supported in 1985 by research into the vastus

lateralis muscle and its buffering capabilities, the results showed that high intensity athletes

such as sprinters had developed greater muscle buffering abilities and intramuscular

carnosine concentrations than low intensity athletes such as marathon runners (Parkhouse,

Mckenzie, Hochachka & Ovalle, 1985). Hence, carnosine’s function as an intracellular pH

buffer and it’s ability to regulate muscular acidosis via artificially induced alkalosis suggest

that it would increase high-intensity exercise capacity and performance (Sale, Saunders &

Harris, 2009). Carnosine’s role as a buffer is likely why it has been shown to increase the

sensitivity of the contractile apparatus to Ca2+ ions involved in the excitation-contraction

coupling process in mammalian skeletal muscle, and contradictory to past studies, the same

study found that carnosine caused a small increase in Ca2+ release when used synergistically

with caffeine (Dutka, Lamboley, McKenna, Murphy, & Lamb, 2011).

Carnosine cannot be supplemented directly, as carnosinase, the enzyme that catalyses

carnosine breakdown, is synthesised in the brain and secreted into the cerebrospinal fluid

where it’s assimilated into the systemic circulation where it would hydrolyse any carnosine

(Schoen, 2003). However, carnosinase is not present within the tissue of skeletal muscles. β-

Alanine has been shown to be the rate limiting factor of carnosine synthesis and unlike

carnosine, hence why it is the supplement of choice rather than L-histidine, the other amino

acid precursor to carnosine. Unlike carnosine, β-alanine avoids enzymatic breakdown before

forming the dipeptide which is stored intramuscularly where there is no carnosinase. Studies

have shown that β-alanine supplementation of between 2-6 milligrams per day can increase

the carnosine content of human muscle tissue by between 20-80% in a dose-dependent

fashion (Derave et al., 2010). In summary, β-alanine’s use as a supplement is a means for

obtaining increased levels of carnosine and it’s effects as an individual peptide have not

been seen to be ergogenic.

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1.6 Aims and ObjectiveThe aim of this double-blind study was to investigate the effects of β-Alanine on anaerobic

endurance in young adult males. Endurance will be measured by determining the number of

repetitions each individual can perform before reaching fatigue-induced failure when

performing both an upper body and lower body compound exercise at a fixed percentage of

their respective bodyweight. Analysis of the hypotheses was completed using One way

analyses of variance (ANOVA) with repeated measures.

1.7 HypothesesNull hypothesis (H0): β-alanine supplementation has no ergogenic effect on the anaerobic

endurance of strength-trained trained adult males.

Alternative hypothesis (HA): β-alanine supplementation increases the anaerobic endurance of

strength-trained trained adult males.

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2. Materials and methods:

2.1 Subjects:

22 healthy male volunteers (mean age ± SD = 20.9±1.1 yrs, height = 176.5±7.0 cm, weight =

80.7±9.6 kg) participated in the research study. The subjects were all recreational weight

lifters all of whom had at least 2 years’ experience training but had not competed in Olympic

weightlifting or powerlifting. Prior to initiation, participants were provided a description of the

study, informing them of all risks associated and were required to provide informed consent

as well as completing and passing a health screen in accordance with the University of

Hertfordshire ethics committee for studies involving the use of human participants. The study

was approved by the University of Hertfordshire ethics committee and was performed on site.

2.2 Supplementation Protocol

None of the participants had supplemented with taurine, β-alanine or creatine of any form in

the 2 months prior to the study, nor had they ever used any form of anabolic steroids or

performance enhancing drugs of a similar nature. Participants weren’t allowed to observe

any irregular diet plans such as veganism, gluten free or ketogenic and weren’t allowed to be

at a considerable caloric deficit or surplus greater than 500kcal at any time during the study.

They were instructed to maintain their current exercise and dietary patterns while abstaining

from any nutritional supplements that they hadn’t provided prior notification of and had

subsequently been deemed acceptable. Each subject was required to continue strength

training 3-5 times per week working all of the major muscle groups. During the study the

participants were required to continue bench pressing and deadlifting without any form of

periodization training specifically designed for peaking, which is defined as “a temporary

state when physical and psychological efficiencies are maximized and where the levels of

technical and tactical preparation are optimal. During this state of training, the individual’s

physiological and anatomical adaptation capacities are at a maximum as well, and the

neuromuscular coordination is perfect” (Bompa & Haff, 1999), this is seen prior to any kind of

weightlifting or strength related competition.

The 22 participants were assigned numbers to ensure privacy, height and weight were

measured prior to baseline testing, between 11 am and 2 pm to avoid shrinkage, water

retention, and food intake being extraneous factors. Height and weight values were used to

calculate the participants’ BMI’s (25.9±2.7), the amount of β-alanine to be consumed daily

(80mg/kg) (6.5±0.8 g/day) as well as the weight being bench pressed (52.4±6.2 kg) (65% of

bodyweight) and deadlifted (96.8±11.5 kg) (120% of bodyweight) during testing. Using the

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Mifflin St. Jeor equation, metabolic rates (2921±283 kcal/day) were calculated for each

subject based on their height, weight, age and estimated weekly exercise. The dosage of

80mg/kg of bodyweight per day was decided upon following the aforementioned observation

of a 2010 study showing that 2-6 mg of carnosine daily can increase carnosine content by up

to 80% in a dose-dependent fashion. The figure of 80mg/kg/day was ideal as it would allow

for maximum carnosine absorption and split over 4 daily doses wouldn’t cause paraesthesia.

2.3 Crossover design

Following baseline testing, the subjects were randomly assigned into two supplement

conditions using a double-blind design, these were a) placebo (PL; 80mg/kg/day

maltodextrin) and b) experiment/β-alanine (E; 80mg/kg/day β-alanine). The supplements

were both provided in tasteless, white powder form and were to be mixed and dissolved in a

minimum of ½ a pint of water (284.1 ml) before ingestion. The supplements were split into 4

even doses to be taken at even intervals throughout the day for a 10 day period, this was to

be noted in food diaries (appendix 3) along with their entire daily diet. The food diaries were

to be kept for 3 days instead of the whole period to ensure participants filled them completely

for a short period that could be extrapolated instead of inconsistently over a long period. All

subjects were again reminded to stick to their usual dietary habits and were also provided

with a list of foods not to consume that were exceptionally high in taurine.

Figure 2.1: A timeline describing the crossover design of the study in chronological order from subject participants acceptance into the study (left) to the completion of the final tests; test 2 (right).

Following the day 10 supplementation period, both groups performed the first set of post

supplementation testing (test). Upon completing the tests all participants were given a

minimum of a one week washout period, this was originally designed to be exactly 7 days for

all participants, however due to unforeseen external circumstances this was not possible

(Mean washout period ± SD= 19.1 ± 8.1 days) . The groups then underwent the “crossover”

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and took the reverse supplement from the first 10 day period, this was again for another 10

days with the same supplement measurements (figure 3).

2.4 Anaerobic testing

Prior to (baseline) and following the supplementation periods, the subjects performed

anaerobic testing which entailed performing two compound weightlifting exercises. The

deadlift and bench press were chosen as they are both compound, thus placing stress on

multiple muscles through the use of two or more joints with the focus being on the opposing

lower and upper body respectively. As earlier stated, participants were to bench press 65%

of their bodyweight followed by deadlift 120% of their bodyweight. Repetitions were to be

performed until failure was reached, at which point they would stop attempting repetitions

and the results would be recorded. For a repetition to be counted as complete, both the

eccentric and concentric portion of the lift were to be completed through the full range of

motion, anything short of this was not included in the result.

In weightlifting, “failure” is defined as “the point of momentary muscular failure where a

repetition fails due to inadequate muscular strength” (Swank & Hagerman, 2010). Additional

measures were inserted for this study to define failure, if the participant completed a

repetition with incorrect form the repetition would not be counted, and if this occurred three

times consecutively due to overexertion then this would be deemed failure, furthermore, if a

participant paused for three or more seconds following the concentric portion of the bench

press or the eccentric portion of the deadlift, this would also be determined as failure, and

finally, if a participant was able to recognise that they had reached muscular failure then they

could signal as such. Subjects were to come in for testing between the hours of 11am and

2pm for all test days and were required to have eaten a meal in the 3 hours prior testing

containing a minimum of 20g of carbohydrates, to avoid dietary habits and circadian rhythm

baring an effect on results.

2.4.1 ParametersDuring the three stages of testing, the parameters measured were; repetitions completed and

time under tension were measured, and as the deadlift does not involve continuous tension

throughout the movement due to the barbell being supported by the floor following the

eccentric portion of the movement, the total time taken was also measured. Time under

tension was measured to ensure participants were performing repetitions at a similar tempo.

2.4.2 TempoDuring testing the participants were told to perform the bench press at a ‘21X0’ tempo where

2 seconds were given to lower the bar in a controlled manner (signified by 2), the pause then

lasted 1 second with the bar touching the chest (1), and following this they were to explode

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the loaded barbell up as fast as possible through the concentric portion (X), this would

constitute one repetition and they would not pause between any repetitions (0). These

instructions were provided to standardise the difficulty of one repetition for each participant

and in theory this would suggest that each repetition would last 3-4 seconds, however to

avoid human errors in controlling the speed of their repetitions, the participants were timed

while performing the bench press. Tempo instructions weren’t provided for the deadlift, the

subjects were advised to perform the concentric as explosively as possible and were

informed that repetitions would not count if the barbell was dropped to the floor without

control once locking out the movement, hence they had to control the eccentric portion of the

deadlift. Once completing each repetition for the deadlift, participants were allowed up to

three seconds if necessary to realign their feet and body with the barbell to ensure injuries

would not occur as a result of lopsided posture while performing the motion.

2.4.3 EquipmentIn addition to tempo standardisation, equipment was uniform for every participant, all lifts

were performed completely unequipped with the exception of an ‘RDX™ cow hide weight

lifting belt’ and ‘Myprotein® liquid chalk’ that were required for use by every participant in

order to lessen the chance of injury and also to avoid fatigue in the extensors and flexors of

the forearm being a limiting factor in the results of a participant.

2.4.4 Warming upParticipants were taken through a stringent warm up routine prior to the bench press that

consisted of performing internal, external and lateral rotations of the rotator cuff for 1 set

each, as well as stretching the pectoralis and deltoids among other shoulder and upper back

muscles by performing behind the neck and overhead stretches for 30 seconds. The internal

and lateral movements are detailed in figure 2.2 and all required use of a dyna-band®.

Following completion of the bench press testing, the subjects underwent a separate warm up

routine to prepare for deadlifting that was aimed at activating both the posterior and anterior

chain of muscles involved in the movement. A hip circle was provided and participants were

made to take 10 steps as far as possible in each direction laterally and longitudinally and this

was followed by one set of barbell good mornings to activate the biceps femoris, gluteus

maximus and thoracolumbar fascia among other secondary muscles used in the deadlift

(figure 2.2). A dyna-band® and hip circle were used as part of a dynamic stretching warm up

to remove any stiffness or immobility as a potential extraneous factor. Dynamic stretching

was favoured over acute static stretching which has been shown to potentially decrease

maximal voluntary contraction (MVC) during the late contraction phase for a short period of

time immediately following the stretching (Morais de Oliveira, Greco, Molina, & Denadai,

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2012). In addition to a physical warm up routine, the temperature of the testing room was

recorded for each day of testing (16.5±1.1°C)

Figure 2.2: (a) Depiction of hip circle in use to prepare for weightlifting by activating the muscles surrounding the synovial joint (sling shot hip circle, 2014). (b) A graphical representation of internal and external rotation of the rotator cuff using a dyna-band (Rotator cuff strain rehabilitation exercises, 2014). (c) The anatomy of a deadlift and the activated muscles in the posterior chain (Thomas, 2008). (d) The anatomy of a deadlift and the activated muscles in the anterior chain (Thomas, 2008).

2.5 Statistical Methods

Standard statistical techniques were used for the calculations of the means, standard

deviations and standard errors of the mean using Microsoft Excel. The Tau-Thompson

technique was used to analyse the data for outliers. A One-way repeated measure analysis

of variance (One-way RM ANOVA) was performed assessing the significance of the

differences in repetitions completed between the (independent variables) baseline,

maltodextrin and β-alanine tests for both the bench press and deadlift. The same One-way

RM ANOVA test was repeated for the same parameters however this time assessing the

significance of the differences in the (independent variables) baseline, test 1 and test 2

experiments, thus assessing for any learning effect that could account for increases in

endurance, which would question the mechanism of any observed effects of the β-alanine.

All statistical tests were performed using IBM SPSS statistics 23. A P value of <0.05 was

used as the criteria for defining statistical significance, Bonferroni correction post hoc

methods were used to analyse the pairwise differences between the groups to ensure.

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3 Data and results:

A one way ANOVA with repeated measures for the baseline, post-placebo and post-β-

alanine tests for both the bench press (P=2.137E-7***) and deadlift (P=1.451E-8***)

repetitions completed produced Greenhouse-Geisser P values of <0.05, the predetermined

criteria for significance, thus allowing the rejection of the null hypothesis; β-alanine has no

effect on anaerobic endurance.

Bonferroni corrected post hoc tests were performed for the bench press and the pairwise

comparisons found that there was a significant increase between both baseline against β-

alanine (P=3.200E-5***, mean difference= 2.773) and post-maltodextrin against post-β-

alanine (P=4.100E-5***, mean difference= 2.681), the mean differences suggesting an

increase in repetitions completed from the baseline and post-maltodextrin tests to the post-β-

alanine tests. The same post hoc tests were repeated for the deadlift and pairwise

comparisons found a significant increase from both the baseline (P= 4.509E-7***, mean

difference= 2.773) and post-maltodextrin (P= 3.851E-7***, mean difference= 2.864) to the

post-β-alanine tests. The alternative hypothesis, stating that β-alanine supplementation

significantly increases anaerobic endurance, was accepted.

There was an increase in the mean repetitions completed for both the bench press and the

deadlift following supplementation (Mean ± SD= 26.05±9.37 BP, 19.86±7.61 DL) with

comparison to the means for the placebo (23.36±8.23 BP, 17.09±6.62 DL) and

baseline(23.27±7.84 BP, 17.09±6.62 DL) test, further supporting the alternative hypothesis.

Mean repetitions completed (values included on graph) for the bench press (figure 3.1) and

deadlift (figure 3.2) were graphed excluding error bars due to the paired nature of the data. A

noticeable difference was visible between the repetitions completed following β-alanine

supplementation in both the bench press and deadlift.

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21.5

22

22.5

23

23.5

24

24.5

25

25.5

26

26.5

23.27 23.36

26.05

Baseline

Post-placebo

Post-β-Alanine

Stage of supplementation

Mea

n nu

mbe

r of B

ench

pre

ss

repe

tition

s com

plet

ed

15.5

16

16.5

17

17.5

18

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19

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17.0917.00

19.86

Baseline

Post-placebo

Post-β-Alanine

Stage of supplementation

Mea

n nu

mbe

r of d

eadl

ift re

petiti

ons

com

plet

ed

Figure 3.2: Bar chart representing the mean deadlift repetitions completed during baseline testing, post-placebo testing and post-β-alanine testing, with mean values labelled.

Page 21 of 84

Figure 3.1: Bar chart representing the mean bench press repetitions completed during baseline testing, post-placebo testing and post-β-alanine testing, with mean values labelled

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3.1 Bench press

The results of the One-way repeated measure analysis of variance (One-way RM ANOVA)

assessing the significance of the differences in bench press repetitions completed for the

baseline, post-placebo and post-β-alanine resulted in Greenhouse-Geisser P=2.137E-7*** as

detailed in appendix 2.1. In summary, the null hypothesis (H0) specifically regarding the

bench press, can be rejected.

Bonferroni corrected post hoc methods for pairwise comparisons of within subject effects for

both baseline against β-alanine (P=3.200E-5***, mean difference= 2.773) and post-

maltodextrin against post-β-alanine (P=4.100E-5***, mean difference= 2.681) indicated

significant differences (Appendix 2.1). The same pairwise comparisons resulted in P=1.000

between baseline and placebo testing. The pairwise comparison values thus allow the

acceptance of the alternative hypothesis.

Post-placebo repetitions completed Post- β-alanine repetitions completed0

5

10

15

20

25

30

35

40

45

13

16

12

17

31

37

16 16

7

10

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43

20

2423

26

32

34

19

21

30

3535

39

31

35

28

31

15

12

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23

19

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30

25

28

17

14

30

34

Subject 1Subject 2Subject 3Subject 4Subject 5Subject 6Subject 7Subject 8 Subject 9 Subject 10Subject 11Subject 12Subject 13Subject 14Subject 15Subject 16Subject 17Subject 18Subject 19Subject 20Subject 21Subject 22

Benc

h pr

ess

repe

tition

s co

mpl

eted

Figure 3.3: Line chart displaying the changes in bench press repetitions completed between post-placebo testing and post-β-alanine testing for each participant, (mean difference ± SD= 2.68±2.23)

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0.00

0.50

1.00

1.50

2.00

2.50

3.00

2.68±0.48

0.09±0.42

2.77±0.48

post-β-alanine vs post-placebo post-placebo vs baselinepost-β-alanine vs baseline

Stage of Supplementation

Mea

n di

ffere

nce

in b

ench

pre

ss re

petiti

ons

Figure 3.4: Bar chart depicting the mean difference in bench press repetitions completed between the 3 tests performed at the different stages of known supplementation. Mean differences displayed ± Standard error of the mean values.

3.2 Deadlift

One-way repeated measure analysis of variance (One-way RM ANOVA) was repeated

assessing the significance of the differences in deadlift repetitions completed for the

baseline, maltodextrin and β-alanine, this time resulting in a within-subject Greenhouse-

Geisser P=1.451E-8*** as detailed in appendix 2.2. H0, for the deadlift, could be rejected.

The alternative hypothesis (HA) was accepted as the Bonferroni corrected pairwise

comparisons between both the baseline (P= 4.509E-7***, mean difference= 2.773) and post-

maltodextrin (P= 3.851E-7***, mean difference= 2.864) testing against the post-β-alanine

were significant, with no significant effect seen between the baseline and post-maltodextrin

repetitions completed (P=1.000) (appendix 2.2)

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-0.50

0.00

0.50

1.00

1.50

2.00

2.50

3.00

3.50

2.86±0.37

-0.09±0.46

2.77±0.36

post-β-alanine vs post-placebo post-placebo vs baseline post-β-alanine vs baseline

Stage of Supplementation

Mea

n di

ffere

nce

in d

eadl

ift re

petiti

ons

Figure 3.5: Bar chart depicting the mean difference in deadlift repetitions completed between the 3 tests performed at the different stages of known supplementation. Mean differences displayed ± Standard error of the mean.

Post-placebo repetitions completed Post-β-alanine repetitions completed0

5

10

15

20

25

30

35

40

12

17

79

17

23

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1517

3638

18

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Subject 1Subject 2Subject 3Subject 4Subject 5Subject 6Subject 7Subject 8Subject 9 Subject 10Subject 11Subject 12Subject 13Subject 14Subject 15Subject 16Subject 17Subject 18Subject 19Subject 20Subject 21Subject 22

Dea

dlift

repe

tition

s co

mpl

eted

Figure 3.6: Line chart displaying the changes in deadlift repetitions completed between post-placebo testing and post-β-alanine testing for each participant, (mean difference ± SD= 2.86±1.73)

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3.3 Chronological analysis: The learning effect

One-way repeated measure analysis of variance (One-way RM ANOVA) was repeated,

assessing the significance of the differences in deadlift repetitions completed for the

chronologically consecutive stages of testing: baseline, test 1 and test 2. This resulted in a

within-subject Greenhouse-Geisser P=0.055 as detailed in appendix 2.3. H0, in this case

stating that for the deadlift, consecutive testing has no effect on the number of repetitions

completed, could not be rejected and thus post hoc testing wasn’t analysed.

16

16.5

17

17.5

18

18.5

19

17.09

18.36

18.50

BaselineTest 1Test 2

Stage of testing

Mea

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r of d

eadl

ift re

petiti

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ompl

eted

Figure 3.7: Mean deadlift repetitions completed during the consecutive stages of testing: baseline testing, test 1 and test 2, with mean values labelled.

The one way ANOVA with repeated measures was used to analyse the results from the

bench press in chronological order, again comparing; baseline, test 1and test 2. This

produced a statistically significant Greenhouse-Geisser of P=0.048 as noted in appendix 2.4,

therefore Bonferroni corrected post hoc testing was used to generate pairwise comparisons.

The pairwise comparisons between test 1 and test 2 inferred that there was no statistical

significance between the repetitions completed during both tests (P=1.000). Baseline testing

was statistically significant in comparison to test 2 (P=0.030) but not statistically significant to

test 1 (P=0.086).

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22

22.5

23

23.5

24

24.5

25

25.5

23.27

24.45

24.95

BaselineTest 1Test 2

Stage of testing

Mea

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r of B

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ompl

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Figure 3.8: Mean bench press repetitions completed during the consecutive stages of testing: baseline testing, test 1 and test 2, with mean values labelled.

3.4 Repetition volume

Mean repetitions were calculated and tabulated for each participant as shown in table 2 for

deadlifts, and table 3 for bench press. In each table, results were ordered numerically from

smallest to largest based on mean repetitions completed across the three stages of

supplementation (Baseline, post-placebo (maltodextrin), experiment (post-β-alanine). These

were then further separated into two groups per exercise, the first being the 1st and 2nd

quartile (highlighted in blue), participants up to the 50th percentile of mean repetitions

completed, the second being the 3rd and 4th quartile, the remaining 50% of participants who

were found to have completed more repetitions than the 1st and 2nd quartile.

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Table 3.1: Mean repetitions completed by each subject for the deadlift across the three stages of supplementation. 1st and 2nd quartile (mean repetitions ± SD= 12.9±2.5) highlighted in blue, 3rd and 4th (23.1±6.3) quartile highlighted in green. Quartiles defined by by mean number of repetitions completed, with 1st and 2nd (lower 50%) having completed less than 3rd and 4th (greater 50%).

DeadliftSubjec

tBaseline repetitions

completedPost-placebo

repetitions completed

Post-β-alanine repetitions completed

Mean

2 8 7 9 85 9 11 12 10. 7

15 9 10 13 10.74 10 12 13 11.7

16 12 11 13 1210 14 11 13 12.721 12 15 16 14.31 15 12 17 14.76 15 15 17 15.7

18 14 17 16 15.720 14 15 18 15.719 16 15 17 1617 16 16 18 16.713 19 17 21 193 20 17 23 20

11 20 19 23 20.78 21 18 24 21

12 23 20 24 22.322 21 22 26 239 25 26 30 27

14 31 32 36 337 32 36 38 35.3

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Table 3.2: The mean repetitions completed by each subject for the bench press across the three stages of supplementation. 1st and 2nd quartile (mean repetitions ± SD= 17.2±4.4) highlighted in blue, 3rd and 4th quartile highlighted in green (31.3±4.5).

Bench PressSubjec

tBaseline

repetitions completed

Post-placebo repetitions completed

Post-β-alanine repetitions completed

Mean Repetitions completed

5 10 7 10 915 13 15 12 13.31 15 13 16 14.72 15 12 17 14.74 14 16 16 15.321 16 17 14 15.710 18 19 21 19.318 19 19 22 207 22 20 24 2217 22 21 23 228 20 23 26 2316 24 24 26 24.720 25 25 28 2619 27 27 30 2814 28 28 31 2922 27 30 34 30.39 29 32 34 31.711 32 30 35 32.313 31 31 35 32.33 33 31 37 33.712 32 35 39 35.36 40 39 43 40.7

Four separate one way ANOVAs with repeated measures were completed comparing the

repetitions completed for both the deadlift and bench press across the 3 stages of

supplementation (Baseline, post-placebo (maltodextrin) and post-β-alanine).

The first and second quartile for the deadlift returned a significant result (P=0.002) as did the

third and fourth quartile (P=1.400E-5***). The pairwise comparisons for the first and second

quartile revealed significant differences between the repetitions completed following β-

alanine supplementation compared to the baseline repetitions completed (P=2.022E-3***,

mean difference= 2.273) and also compared to the placebo repetitions completed

(P=5.634E-3***, mean difference= 1.909). Insignificant differences were observed between

the baseline and placebo repetitions completed (P=1.000). Pairwise comparisons of the third

and fourth quartiles results showed significant differences between the repetitions completed

post β-alanine supplementation against baseline repetitions (P=2.860E-4***, mean

difference= 3.273) and against post-placebo repetitions completed (P=1.200E-5***, mean

difference= 3.818).

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First and second quartile one-way RM ANOVA analysis of the bench press (P=0.049)

suggested significant differences between the supplementation stages, the Bonferroni post

hoc analysis was insignificant for baseline against placebo (P=1.000), baseline against post

post-β-alanine (P=0.118, mean difference= 1.545) and placebo against post-β-alanine

(P=0.168, mean difference= 1.727). Third and fourth quartile analysis was significantly

different between the supplement stages (P=7.696E-7***), when further analysed, the post

hoc results were significant between the baseline and post-β-alanine tests (P=3.800E-5***,

mean difference= 4.000) and between the post-placebo and post-β-alanine tests (P=5.000E-

6***, mean difference= 3.636). The repetitions between the baseline test and post-placebo

were statistically insignificant (P=1.000).

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4. Discussion:

β-alanine was found to have an ergogenic effect on anaerobic endurance, this had previously

been surmised following a 2009 literature review concerning β-alanine (Giannini Artioli,

Gualano, Smith, Stout, & Herbert Lancha, 2009). The complementary constituent to L-

histidine in the dipeptide carnosine; β-alanine has been shown to be the rate limiting factor in

carnosine synthesis (Sale, Saunders & Harris, 2009). Carnosine’s biochemical makeup,

containing an imidazole ring side chain as well as a carboxyl and amino group, with pKa’s, of

6.83, 2.77 and 9.63 respectively, allows it to function as a pH buffer in vivo (Smith, 1938).

However, due to the presence of the degradative carnosinase throughout various tissue of

the human body as well as the sera, it cannot be consumed without being broken down

before fulfilling its role as a buffer (Schoen, 2003). Hence, β-alanine could be supplemented

to increase the carnosine concentrations of the skeletal muscle in an attempt to regulate the

pH changes observed as a result of acidosis during intense anaerobic exercise. This study

attempted to investigate the effects of β-alanine supplementation on intramuscular buffering

during anaerobic glycolysis.

Significantly increased repetitions (P<.05) were observed following β-alanine

supplementation in both exercises, the bench press and deadlift. This supported the

hypothesis (HA) that β-alanine supplementation increases anaerobic endurance and also

suggests β-alanine’s ergogenic effects aren’t selective for the performance of a specific

region of the body for any physiological reasons.

4.1 ATP:CP to Anaerobic glycolysis

During testing, as repetitions and subsequently intensity of muscle contractions increased,

the emphasis in providing energy for the contractions shifted between different energy

systems, the majority of the ATP immediately provided in extremely short term, maximal

effort exercises is provided by the ATP:CP (phosphagen) system, however no matter the

intensity of the exercise, the phosphagen system would’ve been activated at the onset of

exercise due to its ability to rapidly produce energy compared to the glycolytic and oxidative

pathways. Stores of intramuscular phosphocreatine, the molecule hydrolysed to provide the

energy for ATP synthesis in the phosphagen system, are relatively small and will only last up

to around 10 seconds, depending on the person and the exercise being performed, at which

point glycolysis becomes the predominant supply of ATP for energy production until aerobic

sources of ATP are required, thus explaining why creatine supplementation that would

maximise stores of phosphocreatine was disallowed among all participants as the pathway

precedes that of the glycolytic, this is in addition to evidence of it having shown synergistic

effects when combined with β-alanine (Hoffman et al., 2006).

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4.2 Exercise volume and ATP production

Relating the energy systems to the observed results of the study, the exercises were

performed at fixed weights but varied across a range of repetitions between 7 and 43 with

the shortest time for completion of a testing set (measured as time under tension, therefore

resetting of foot position wasn’t included in time for deadlifts) measuring 18 seconds and the

longest time measuring 97 seconds. The lower limits of these parameters exceeded the

lactate threshold and surpassed the time in which the phosphagen system can be the sole

energy supply for muscle contraction, therefore anaerobic glycolysis would’ve been used to

provide ATP for hydrolysis, releasing energy for the contraction of the muscles needed to

perform the exercises. Assuming this was true, muscle fatigue and subsequent failure

would’ve been at least partially resultant of the acidosis generated via anaerobic glycolysis.

Therefore failure in the subject’s would’ve been brought on by the H+ regulated inhibition of

Ca2+ binding to troponin C in myofibril contraction (Ball, Johnson, & Solaro, 1994). The

increased mean repetitions observed from the baseline and post-placebo testing to post-β-

alanine testing (figures 3.4 & 3.5) suggests that as intramuscular carnosine concentration

increased following β-alanine supplementation, the buffering potential of the skeletal muscles

involved in the exercise also increased, slowing down the onset of troponin C-mediated

excitation-contraction coupling inhibition also increased, hence the ability to contract the

muscles for long enough to continue for more repetitions before reaching failure.

Following this, the participants’ results were divided into groups based on the quantity

repetitions completed for each exercise. The individuals completing less repetitions prior to

supplementation saw a less significant increase in repetitions post-β-alanine for both the

bench press and deadlift (table 2 & 3). As earlier stated, as repetitions performed increases,

the ATP:CP system becomes less of a factor in energy production and anaerobic glycolysis

becomes more of a factor, thus, those individuals who were ‘stronger’ pound for pound and

could lift a fixed fraction of their body weight for more repetitions than their counterparts,

placed more stress on the anaerobic glycolytic pathway therefore causing a marked

decrease in pH associated with lactate and particularly H+ ion build up from ATP hydrolysis

(Robergs, Ghiasvand, & Parker, 2004). This increased reduction in pH in turn increased the

significance of carnosine’s role as a pH buffer compared to the ‘weaker’ participants who

placed less stress on their anaerobic glycolytic energy system, thus requiring less pH

buffering and instead achieving failure through other forms of muscle fatigue particularly Pi

accumulation from the ATP:CP system. This is supported by figure 4.1 below, which shows

Pi actually inhibiting Ca2+ release from rat skinned muscle fibres, which as earlier stated is

released to bind troponin C and induce myofibril contraction.

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Figure 4.1: Force records from the SR of rat skinned muscle fibres. Caffeine was applied to release SR Ca2+ producing the above contractures; thus the size of the contracture is an indication of the Ca2+ released in the SR. The middle recording follows the muscle be being exposed to 50 mm Pi for 20 s prior to washing off and caffeine application (Fryer, Owen, Lamb, & Stephenson).

The effect of being ‘stronger’ potentially had a two-fold effect in increasing carnosine’s

intramuscular effects. Individuals who were stronger would in theory contain greater amounts

of muscle, and also an altered muscle composition due to chronic training, which will be

discussed, thus allowing a greater amount of carnosine to be stored, therefore allowing for a

greater buffering effect.

However, it should be noted that these observations, while statistically significant

(appendices 2.5-2.8) could have been due to numerous factors, “high responders” and “low

responders” to β-alanine supplementation have been observed, this has been inconclusively

attributed to diet, muscle composition and even style of training (Giannini Artioli, Gualano,

Smith, Stout, & Herbert Lancha, 2009).

4.3 Muscle composition

The increases in repetitions post-supplementation observed in the deadlift compared to the

bench press, were likely due to the concentration of fast twitch, type II, muscle fibres within

the muscles involved. There are large amounts of type IIb (fast twitch glycolytic) muscle

fibres within the vastus lateralis, particularly in men. These fibres predominantly contain the

largest amounts of carnosine thus suggesting β-alanine supplementation will aid anaerobic

endurance within these muscles (table 4.1), other muscles used in the deadlift including the

tibialis anterior, biceps femoris and gastrocnemius also generally contain a large amount of

carnosine comparative to other muscles of the body (Crill, et al., 1999) (Culbertson, Kreider,

Greenwood, & Cooke, 2010).

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Table 3: Fiber types, carnosine content, and buffering capacity of the middle gluteal muscle of thoroughbred horse (Sewell, Harris, Marlin & Dunnett, 1992).

In contrast to the muscle composition being a factor, the bench press repetitions completed

were noticeably higher on average (26.0±1.99) compared to the deadlift (19.9±1.62 DL),

which would suggest greater endurance was required, however the times spent under

tension (appendix 4.3&4.4) were within the same range and most times weren't in the range

of 60-240 seconds which has been suggested as the ideal time for β-alanine to have a

positive ergogenic effect on endurance (Hobson, Saunders, Ball, Harris, & Sale, 2012).

4.4 Future challenges

The use of maltodextrin powder as placebo for β-alanine powder potentially disturbed the

double blind nature of the trial; β-alanine can cause paraesthesia via the binding and

activation of the MrgprD receptor within sensory neurons, however, maltodextrin cannot

cause paraesthesia, in an attempt to counteract this, the daily dosage of β-alanine was taken

in 4 smaller doses throughout the subjects’ day, but in future pre-mixing the powder into

solution would avoid any potential issues including appearance (Liu et. Al, 2012).

The use of height and weight to calculate BMI as a parameter to describe the size of the

participants is an outdated method as it gives very little indication of whole body composition.

Hydrostatic weighing and air displacement plethysmography would be ideal methods for

ascertaining bone mass, body fat percentage (including visceral fat), total water mass,

muscle mass (segmental and overall) and fat free mass index. In future bioelectrical

impedance could be used as a cheaper alternative however as of now it can’t account for the

physiological differences of various ethnicities when making measurements (Dehghan &

Merchant, 2008).

The biological parameters particularly overall and segmental muscle mass, would’ve been

useful in analysing the results of the testing. Carnosine is stored within skeletal muscle,

particularly fast twitch, and the brain, however β-alanine supplementation has been shown

not to affect carnosine concentrations within the brain (Solis et al., 2015). It would be

Page 33 of 84

Table 4.1: Fiber types, carnosine content, and buffering capacity of the middle gluteal muscle of thoroughbred horse (Sewell, Harris, Marlin & Dunnett, 1992).

Page 34: The ergogenic effect of beta alanine on anaerobic endurance 13035885

interesting to observe if a relationship existed between individuals with larger muscle masses

and the added endurance benefits received from supplementation of β-alanine in comparison

with those holding less muscle mass.

With the goal of determining the severity of acidosis induced immediately post-exercise,

measuring blood lactate concentrations directly following the completion of the anaerobic

tests performed would be ideal. As aforementioned, lactate build up is a very reliable marker

in the diagnosis of intramuscular acidosis, this would’ve been a more direct method to

assess the ergogenic affects β-alanine supplementation is having on anaerobic endurance

via carnosine’s role as a pH buffer. Increased lactate concentration in individuals post-

supplementation at the time of failure would indicate carnosine mediated buffering has

helped combat the effects of acidosis, thus measuring lactate would aid in conclusively

determining that the effects of the increased repetitions are due to carnosine’s mechanism of

buffering.

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Conclusion:

β-alanine was found to impart an ergogenic effect across the 22 participants, significantly

increasing the number of repetitions completed and therefore anaerobic endurance in both

compound exercises, the deadlift and the bench press. The null hypothesis that β-alanine

would have no ergogenic effect on anaerobic endurance was disproved by One way ANOVA

with repeated measures results stating significance (P<0.005) between results pre and post

supplementation. The alternative hypothesis that β-alanine will cause an increase in

anaerobic endurance was supported by post hoc pairwise comparisons indicating

significance between both the post-placebo and baseline repetitions completed against the

post- β-alanine testing. This suggests that carnosine concentrations were increased and

induced alkalosis to buffer against fatigue-inducing acidosis. However, it should be noted that

in future it would be advantageous to measure post-exercise lactate concentrations to further

validate the mechanism of β-alanine’s ergogenic effects.

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5. References:

Ball, K. L., Johnson, M. D., & Solaro, R. J. (1994). Isoform specific interactions of troponin I and troponin C determine pH sensitivity of myofibrillar Ca2+ activation. Biochemistry, 33(28), 8464-8471. doi:10.1021/bi00194a010

Bompa, T. O., & Haff, C. G. (1999). Periodization: Theory and methodology of training (5th ed.). Champaign, IL: Human Kinetics.

Brooks, G. A., Dubouchaud, H., Brown, M., Sicurello, J., & Butz, C. (1999). Role of mitochondrial lactate dehydrogenase and lactate oxidation in the intracellular lactate shuttle. Proceedings of the National Academy of Sciences of the United States of America, 96(3), 1129-1134. doi: 10.1073/pnas.96.3.1129

Chin, E. R., & Allen, D. G. (1998). The contribution of pH-dependent mechanisms to fatigue at different intensities in mammalian single muscle fibres. The Journal of Physiology, 512(3), 831-840. doi:10.1111/j.1469-7793.1998.831bd.x

Crill, M. T., Staron, R. S., Hagerman, F. C., Hikida, R. S., Hostler, D., Ragg, K. E., … Toma, K. (1999). Fiber type composition of the Vastus lateralis muscle of young men and women. Medicine & Science in Sports & Exercise,31(Supplement), S328. doi:10.1097/00005768-199905001-01645

Culbertson, J. Y., Kreider, R. B., Greenwood, M., & Cooke, M. (2010). Effects of Beta-Alanine on Muscle Carnosine and Exercise Performance: A Review of the Current Literature.Nutrients, 2(1), 75-98. doi:10.3390/nu2010075

Dehghan, M., & Merchant, A. T. (2008). Is bioelectrical impedance accurate for use in large epidemiological studies? Nutrition Journal, 7(1), 26. doi:10.1186/1475-2891-7-26

Derave, W., Everaert, I., Beeckman, S., & Baguet, A. (2010). Muscle Carnosine Metabolism and β-Alanine Supplementation in Relation to Exercise and Training. Sports Medicine,40(3), 247-263. doi:10.2165/11530310-000000000-00000

Dutka, T. L., Lamboley, C. R., McKenna, M. J., Murphy, R. M., & Lamb, G. D. (2011). Effects of carnosine on contractile apparatus Ca2+ sensitivity and sarcoplasmic reticulum Ca2+ release in human skeletal muscle fibers. Journal of Applied Physiology, 112(5), 728-736. doi:10.1152/japplphysiol.01331.2011

Giannini Artioli, G., Gualano, B., Smith, A., Stout, J., & Herbert Lancha, A. (2009). The role of β-alanine supplementation on muscle carnosine and exercise performance. Medicine & Science in Sports & Exercise, 42(6). doi:10.1249/mss.0b013e3181c74e38

Hobson, R. M., Saunders, B., Ball, G., Harris, R. C., & Sale, C. (2012). Effects of β-alanine supplementation on exercise performance: a meta-analysis. Amino Acids, 43(1), 25-37. doi:10.1007/s00726-011-1200-z

Hoffman, J. R., Ratamess, N. A., Kang, J., Mangine, G., Faigenbaum, A. D., & Stout, J. R. (2006). Effect of Creatine and β -Alanine Supplementation on Performance and Endocrine Responses in Strength/Power Athletes. Medicine & Science in Sports & Exercise, 38(Supplement), S126. doi:10.1249/00005768-200605001-00581

Liu, Q., Sikand, P., Ma, C., Tang, Z., Han, L., Li, Z., … Dong, X. (2012). Mechanisms of itch evoked by β-alanine. The Journal of Neuroscience : The Official Journal of the Society for Neuroscience, 32(42), 14532–14537. http://doi.org/10.1523/JNEUROSCI.3509-12.2012

Maclaren, D. P., Gibson, H., Parry-Billings, M., & Edwards, R. H. (1989). A Review of Metabolic and Physiological Factors in Fatigue. Exercise and Sport Sciences Reviews,16, 29-66. doi:10.1249/00003677-198900170-00005

McLester, J. R. (1997). Muscle contraction and fatigue. The role of adenosine 5'-diphosphate and inorganic phosphate. Sports Medicine, 23(5), 287-305. doi:10.2165/00007256-199723050-00003

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Morais de Oliveira, A. L., Greco, C. C., Molina, R., & Denadai, B. S. (2012). The Rate of Force Development Obtained at Early Contraction Phase Is Not Influenced by Active Static Stretching. Journal of Strength and Conditioning Research, 26(8), 2174-2179. doi:10.1519/jsc.0b013e31823b0546

Pate, E., Bhimani, M., Franks-Skiba, K., & Cooke, R. (1995). Reduced effect of pH on skinned rabbit psoas muscle mechanics at high temperatures: implications for fatigue. The Journal of Physiology, 486(3), 689-694. doi:10.1113/jphysiol.1995.sp020844

Rotator cuff strain rehabilitation exercises. (2014). Retrieved from http://www.summitmedicalgroup.com/library/adult_health/sma_rotator_cuff_injury_exercises/

Robergs, R. A., Ghiasvand, F., & Parker, D. (2004). Biochemistry of exercise-induced metabolic acidosis. AJP: Regulatory, Integrative and Comparative Physiology, 287(3), R502-R516. doi:10.1152/ajpregu.00114.2004

Sale, C., Saunders, B., & Harris, R. C. (2009). Effect of beta-alanine supplementation on muscle carnosine concentrations and exercise performance. Amino Acids, 39(2), 321-333. doi:10.1007/s00726-009-0443-4

sling shot hip circle. (2014). Retrieved from http://www.rogueapo.com/sling-shot-hip-circle

Schoen, P. (2003). Serum Carnosinase Activity in Plasma and Serum: Validation of a Method and Values in Cardiopulmonary Bypass Surgery. Clinical Chemistry, 49(11), 1930-1932. doi:10.1373/clinchem.2003.019398

Sewell, D. A., Harris, R. C., Marlin, D. J., & Dunnett, M. (1992). Estimation of the carnosine content of different fibre types in the middle gluteal muscle of the thoroughbred horse. The Journal of Physiology, 455, 447–453.

Solis, M. Y., Cooper, S., Hobson, R. M., Artioli, G. G., Otaduy, M. C., Roschel, H., … Sale, C. (2015). Effects of Beta-Alanine supplementation on brain Homocarnosine/Carnosine signal and cognitive function: an exploratory study. PLOS ONE, 10(4), e0123857. doi:10.1371/journal.pone.0123857

Stackhouse, S. K., Reisman, D. S., & Binder-Macleod, S. A. (2001). Challenging the role of pH in skeletal muscle fatigue. Journal of the American Physical Therapy Association, 81(12), 1897-1903. Retrieved from http://ptjournal.apta.org/content/81/12/1897

Taylor, A. D., Humphries, B., Smith, P., & Bronks, R. (1997). Electrophoretic Separation of Myosin Heavy Chain Isoforms in the Human M. Vastus Lateralis: References to Reproducibility and Relationships with Force, Electromechanical Delay, Fibre Conduction Velocity, Endurance and Electromyography. Archives of Physiology and Biochemistry, 105(1), 10-18. doi:10.1076/apab.105.1.10.13142

Thomas, G. (2008). Deadlift rear anatomy. Retrieved from http://www.freefitnessguru.com/Anatomy/Deadlift_Rear_Anatomy.html

Thomas, G. (2008). Deadlift front anatomy. Retrieved from http://www.freefitnessguru.com/Anatomy/Deadlift_Rear_Anatomy.html

Van Meerhaeghe, A., & Velkeniers, B. (2005). Lactate production and exercise-induced metabolic acidosis: guilty or not guilty? European Respiratory Journal, 26(4), 744-744. doi:10.1183/09031936.05.00059005

WebMD printable food & fitness journal. (2008, November 26). Retrieved from http://www.webmd.com/diet/printable/food-fitness-journal

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Appendix1. Ethics forms

EC1:

UNIVERSITY OF HERTFORDSHIRE

FORM EC1: APPLICATION FOR ETHICAL APPROVAL OF A STUDY INVOLVING HUMAN PARTICIPANTS(See Guidance Notes)

Office Use only Date Received by Clerk:

Expedited ReviewApproved by Reviewer 1 (sign & date) Approved by Reviewer 2 (sign & date)

Further Action: (tick appropriate box and provide details)Request Further Information Details:

Refer for Substantive ReviewRefer for Full review

RejectSubstantive ReviewApproved by Reviewer 1 (sign & date) Approved by Reviewer 2 (sign & date)

Approved by Reviewer 3 (sign & date) Approved by Reviewer 4 (sign & date)

Further Action: (tick appropriate box and provide details)Request Further Information Details:

Refer for Full reviewReject

Full ReviewRequest Further Information Details:

Reject

CONFIRMATION OF APPROVAL[To be completed by the Chairman or Vice-Chairman of the relevant ECDA, or by the Chairman of the University Ethics Committee – (see GN 2.1.4)]

I confirm that this application has been approved by or on behalf of the committee named below.

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Relevant ECDA:

Science & Technology

[email protected]

Health & Human Sciences

X

OFFICE USE ONLY

Protocol Number:

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Name/Sign…………………………………………………………Date……………………..

Name of committee ……………………...

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DECLARATIONS1 DECLARATION BY APPLICANT (See GN 2.1.3)

1.1 I undertake, to the best of my ability, to abide by UPR RE01, ‘Studies Involving the Use of Human Participants’, in carrying out the study.

1.2 I undertake to explain the nature of the study and all possible risks to potential participants, to the extent required to comply with both the letter and the spirit of my replies to the foregoing questions (including information contained in Appendices 1 & 2).

1.3 Data relating to participants will be handled with great care. No data relating to named or identifiable participants will be passed on to others without the written consent of the participants concerned, unless they have already consented to such sharing of data when they agreed to take part in the study.

1.4 All participants will be informed (a) that they are not obliged to take part in the study, and (b) that they may withdraw at any time without disadvantage or having to give a reason.

(NOTE: Where the participant is a minor or is otherwise unable, for any reason, to give full consent on their own, references here to participants being given an explanation or information, or being asked to give their consent, are to be understood as referring to the person giving consent on their behalf. (See Q 19; also GN Pt. 3, and especially 3.6 & 3.7))

Enter your name here: Ernest Didehvar-Sadr………………………..Date: 11/12/15………….

2 GROUP APPLICATION

(If you are making this application on behalf of a group of students or staff, please complete this section as well)

I confirm that I have agreement of the other members of the group to sign this declaration on their behalf

Enter your name here ……………………………………………………. Date …………………

3 DECLARATION BY SUPERVISOR (see GN 2.1.3)

I confirm that the proposed study has been appropriately vetted within the School in respect of its aims and methods as a piece of research; that I have discussed this application for Ethics Committee approval with the applicant and approve its submission; that I accept responsibility for guiding the applicant so as to ensure compliance with the terms of the protocol and with any applicable ethical code(s); and that if there are conditions of the approval, they have been met.

Enter your name here………………………………………………………...Date………………

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PERMISSIONS

DECLARATION BY APPLICANT (please refer to Question 7 and GN 2.2.1)

Please select ONE of the following statements and delete the rest:

I have yet to obtain permission but I understand that this will be necessary before I commence my study and that the original copies of the permission letters must be verified by my supervisor by the time I submit my results (please delete if not applicable).

Enter your name here Ernest Didehvar-Sadr………………………. Date 15/12/15…………

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Instructions for Applicants

Applicants are advised to read the Guidance Notes before completing this form. Use of this form is mandatory [see UPR RE01, SS 7.1 to 7.3].

Approval must be sought and granted before any investigation involving human participants begins [UPR RE01, S 4.4(iii)].

Abbreviations

GN=Guidance Notes UPR=University Policies & RegulationsQ=Question S=Section SS=Sections Pt =Part

PLEASE NOTE: Where alternative answers are offered, put a cross in the appropriate box.

For example: YES

Where a “write in” answer is requested, begin in the space provided below the question and continue as necessary. All questions must be answered. Please answer in BLACK INK.

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X

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1. THE STUDY

Q1. Please give the title (or provisional title) of the proposed study. (NB – you will be asked for further details later) The effects of daily consumption of beta alanine on anaerobic strength

2. THE APPLICANT

Q2. Please answer either Q2.1 or Q2.2 by providing the information requested. Q2.1 should be answered by individual applicants, both staff and students, who require protocol approval for work which they themselves intend to carry out. Q2.2 should be answered by academic staff requiring approval for standard protocols governing classroom practical work (or equivalent work) to be carried out by a specified group of students. (See GN 2.2.2)

Q2.1. Name of applicant/(principal) investigator

Ernest Didehvar-Sadr………………………………………………………………………….

Student registration number (or staff number for staff application)

13035885………………………………………………………………………….

Email address

[email protected]………………………………………………………………………….(in the case of a group application, please list the names, registration numbers and email addresses of all members of the group, starting with the lead applicant)

Status:

(a) undergraduate

(b) postgraduate (taught/research)

(c) academic staff

(d) other - please give details here

School/Department: School of life and medical sciences

If application is from member of staff or student NOT based at University of Hertfordshire, please give the name of the institution:

Name of Programme (eg BSc (Hons) Computer Science)BSC (Hons) Biomedical ScienceModule name and module code

6LMS0008-0906-ProjectName of supervisor: Dr. Fang Lou

Supervisor’s contact details (email, extension number)Email: [email protected] Extension: 4532Name of Module Leader if applicant is undertaking a taught programme/module: Kathleen Graeme-Cook

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X

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Name of Programme Tutor: Dr. Jen Young

Q2.2. Class Protocol Applications Only.(do NOT use this section if you are a group of students undertaking a joint project, instead, complete Q2.1 and list the names of all students involved in the project, together with their student registration numbers. It is assumed that just one member of staff will be responsible for supervision.)

Name of applicant/(principal) investigator (member of staff)

School/Department

Programme of study or award (e.g. BSc/MA)

Module Title and Code

Year/group to be governed by the protocol

Number of students involved in study per academic year

Programme Tutor (if different from the applicant)

University of Hertfordshire E-mail address

Please note: Risk Assessment Form EC5 (or subject specific risk assessment) is mandatory for all Class Protocol Applications and must accompany this application.

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3. DETAILS OF THE PROPOSED STUDY

Q3.1 Is it likely that your application will require approval by a National Research Ethics Service (NRES) ethics committee whereby completion of an IRAS form would be required? (See GN 2.2.3)

If you are unsure whether your application should be referred to a National Research Ethics Service (NRES) ethics committee, please use the NHS decision tool. The ECDA clerks have the details. Should you receive an indication that it is not necessary to submit your application to an NRES ethics committee, or if the application is being submitted to NRES by a collaborating institution, please continue to complete Form EC1.

YES NO

(If YES, please answer 3.2 and 3.3)(If NO, please continue on to Q4)

Q3.2 Please confirm whether your research involves any of the following:

Exposure to any ionising radiation

NHS or Social Care patients

NHS or Social Care staff*Note, it is not always necessary to refer studies involvingthis group of participants to the NRES for approval: studentsshould consult their supervisor concerning use of NHS decision tool

Clinical Trial of an Investigational Medical product

Clinical Trial of a Medical Device

Exposure to any ionising radiation

Adults who lack the capacity to consent

Human Tissue (see GN 2.2.3)

Q3.3 If your study is considered to be a Clinical Trial of an Investigational Medical Products (CTIMP) or Clinical Trial of a Medical Device, please indicate if the study involves any of the following categories: (See GN 2.2.3)

Children under 5

Pregnant women

A trial taking place overseas

A trial with more than 5,000 participants

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X

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If you have answered ‘Yes’ to any of the above questions in (b), you will most probably require NRES ethics committee approval and a sponsor. You will need to apply for NRES ethics committee approval using an IRAS form. For University of Hertfordshire sponsorship, you will need to complete Form SP1, which is obtainable as well as further advice regarding IRAS from [email protected].

DO NOT complete this form any further but submit it to your relevant ECDA now. Please note, you will be issued with a UH Protocol Number but this will not be valid until you have sent your relevant ECDA a copy of your NRES ethics committee approval letter and copy of the synopsis of the study.

Q4. Please give a short synopsis of your proposed study; stating its aims and highlighting where these aims relate to the use of human participants. (See GN 2.2.4)Please enter details here. The aims of this study will be to determine whether beta alanine has an effect on strength in trained males who have never supplemented with the aforementioned amino acid before. Beta alanine is a modified form of the amino acid; alanine, it is an ingredient in many pre workout supplements that are designed to give weightlifters increased energy and strength to help them lift the heaviest weights possible. Studies have shown that it has a positive effect on muscular endurance and potentially also anaerobic running capacity when supplemented daily. I wish to determine whether this affect is transferrable to strength as well as endurance as it is used in many supplements that are aimed at increasing muscular strength.

Q5. Please give a brief explanation of the design of the study and the methods and procedures used, highlighting in particular where these involve the use of human participants. You should clearly state the nature of the involvement the human participants will have in your proposed study and the extent of their commitment. Thus you must complete and attach the Form EC6 (Participant Information Sheet) (see Appendix 2). Be sure to provide sufficient detail for the Committee to be clear what is involved in the proposed study, particularly in relation to the human participants. (See GN 2.2.5)

I will pre-screen potential participants to ensure they aren’t supplementing with anything that could potentially affect the study, such as beta alanine or taurine. In addition to this I will perform a health screen to ensure they have no underlying health conditions, allergies or injuries In this project, which will be performed in a double blind format, participants will be separated into groups. Groups will be randomly assigned and they will be as follows: group 1 will consist of participants will be given a 2 week supplementation plan for beta alanine that will consist of a daily dose of beta alanine equivalent to 100mg/kg/day spread out across 4 doses a day Group 2 will be given a placebo, 4 times a day for 2 weeks. Following the opening 2 week period each group will test their 5 rep maximums for deadlifts and bench press with the results being privately stored. The study will then be crossed over and each group will swap for 2 weeks and then tested again. In order to ascertain the doses required for each participant they will come in prior to the study to be weighed, and they will also need to perform a baseline for the deadlift and bench press 5 rep maxes, they will also begin taking a food diary so that their diets can be analysed. Prior to consumption they will perform a bench press and deadlift for sets of 5 repetitions at maximum possible weight and then following the supplementation plan provided (either placebo or non-placebo) for 2 weeks will perform the same exercises observing if they are able to lift a heavier weight for the same number of

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repetitions, form will be monitored and standardised. If participants aren’t dedicated to the supplementation required then they will be removed from the study, other than this no other commitment is foreseeable at this time.

Q6.1 Please give the starting date 08/2/2016

Q6.2 Please give the finishing date. 30/3/2016

(For meaning of “starting date” and “finishing date”, see GN 2.2.6)

Q7. Where will the study take place? University of Hertfordshire physiology labs (H260) (G111)

Please refer to the Guidance Notes (GN 2.2.1) which set out clearly what permissions are required; ensure that you complete the Permissions box near the front of this application form and indicate in Appendix 2 (last page of this application form) which permissions you are attaching to the application.

Please enter details here.

Q8. It might be appropriate to conduct a risk assessment of the proposed location for your study (in respect of hazards/risks affecting both the participants and/or investigators) – this would be particularly relevant for off-campus locations but please consider potential hazards on-campus as well (Question 11 also refers). Please use Form EC5 which is an example of a risk assessment OR use a subject specific risk assessment form provided by your School or Supervisor (See GN 2.2.7 and Section 4 of the Guidance Notes).

If you do not consider it is necessary to make a risk assessment, please give your reasons:

Q9.1 Will anyone other than yourself and the participants be present with you when conducting this study? (See GN 2.2.8)

YES NO

If YES, please state the relationship between anyone else who is present other than the applicant and/or participants (e.g. health professional, parent/guardian of the participant)Lab supervisors

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Spotters

Q9.2 Will the proposed study be conducted in private? If NOT, what steps will be taken to ensure confidentiality of the participants’ information. (See GN 2.2.8)

Yes, confidentiality will be maintained, data will be stored privately, participants will be given numbers instead of having their names used, however the study will be conducted in the presence of others.

4. HARMS, HAZARDS & RISKS

Q10. Will this study involve invasive procedures on the human participants? (See GN 2.2.9)

Yes No

(If YES, please fill out Appendix 1 – Increased Hazards and Risks.Once this is complete, move on to Q15)

(If NO, answer Q11, Q12, Q13 and Q14)

Q11, Q12, Q13 & Q14 - NON INVASIVE STUDIES ONLY

Note: You are advised to read GN 2.2.10, 2.2.11, 2.2.12 & 2.2.13 carefully before you answer the following questions.

Q11. Are there potential hazards to participant(s) and/or investigator(s) from the proposed study? (See 2.2.10)

YES NO

If YES,

Indicate their nature here.

As a result of the exercises being performed, physical injury may occur to the participants, either via a soft tissue strain or via the mishandling of heavy weights, mishandling of the weights may also result in injury to myself or anyone else present. Additional hazards may arise from consumption of the supplements, Side effects of beta alanine have been: mild paraesthesia (common) and taurine deficiency.

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X

X

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Indicate here what precautions will be taken to avoid or minimise any adverse effects.Participants will be taken through a series of warm up exercises to minimise the risk of soft tissue injuries and in the case of potential side effects from the supplements, should the participants feel any serious side effects they will be removed from the study and supplementation will stop immediately.

Q12. Will or could the study cause discomfort or distress of a mental or emotional character to participants and/or investigator(s)? (See GN 2.2.11)

YES NO

If YES,

Indicate its nature here.

Indicate here what precautions will be taken to avoid or minimise such adverse effects.

Q13. Will or could medical or other aftercare and/or support be needed by participants and/or investigator(s) as a result of the study? (See GN 2.2.12)

YES NO

Q14 Please describe in appropriate detail what you would do should the adverse effects or events which you believe could arise from your study, and which you have mentioned in your replies to the previous questions, occur.(See UPR RE01, S 2.3 (ii) and GN 2.2.13)

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5. ABOUT YOUR PARTICIPANTS

Q15. Please give a brief description of the kind of people you hope/intend to have as participants, for instance, a sample of the general population, University students, people affected by a particular medical condition, children within a given age group, employees of a particular firm, people who support a particular political party, and state whether there are any upper or lower age restrictions. University student males between the age of 18 and 25 who have never previously supplemented with beta alanine and don’t currently supplement with taurine either.

Q16. Please state here the maximum number of participants you hope will participate in your study.Please indicate the maximum numbers of participants for each method of data collection.

24 (8 placebo, 8 beta alanine long term, 8 beta alanine pre workout)

Q17. By completing this form, you are indicating that you are reasonably sure that you will be successful in obtaining the number of participants which you hope/intend to recruit. Please outline here your recruitment (sampling) method and how you will advertise your study. (See GN 2.2.14)

Self-recruitment (ideally will get enough from this alone), studynet advertising

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6.CONFIDENTIALITY AND CONSENT

[For guidance on issues relating to consent, see GN 2.2.15 & Pt. 3.]

Q18. Is it intended to seek informed consent from the participants?

YES NO

(See UPR RE01, S 2.3 & 2.4 and GN 3.1)

If YES, please attach a copy of the Consent Form to be used (See Form EC3 & EC4 for reference and GN 3.2), or describe here how consent is to be obtained and recorded. The information you give must be sufficient to enable the Committee to understand exactly what it is that prospective participants are being asked to agree to. If consent is implicit or to be provided by someone other than the participant (such as a parent or guardian), please provide details here.

If NO, please explain why it is considered unnecessary or impossible or otherwise inappropriate to seek informed consent.

Q19. If the participant is a minor (under 18 years of age), or is otherwise unable for any reason to give full consent on their own, state here whose consent will be obtained and how? (See especially GN 3.6 & 3.7)

No minors will be used in the study

Q20. Are personal data of any sort (such as name, age, gender, occupation, contact details or images) to be obtained from or in respect of any participant? (See GN 2.2.16) (You will be required to adhere to the arrangements declared in this application concerning confidentiality of data and its storage. The Participant Information Sheet (EC6 or equivalent) must explain the arrangements clearly.)

YES NO

If YES,

Give details here of personal data to be gathered, and indicate how it will be stored. Age, weight, height, gender, underlying health issues

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X

X

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State here what steps will be taken to prevent or regulate access to personal data beyond the immediate investigative team, as indicated in the Participant Information Sheet?

All data collected will be electronically stored and will only be accessible via password that only I will know, rather than using names each participant will be assigned a 6 digit number that they will be identified with and any unique information that could give away their identity.

Indicate here what assurances will be given to participants about the security of, and access to, personal data, as indicated in the Participant Information Sheet.

Aside from the data including no names or any personal information that would give away the identity of the individuals, and being password protected by myself, it will be destroyed as soon as the study is complete and is no longer needed for review.

State here, as far as you are able to do so, how long personal data collected during the study will be retained, and what arrangements have been made for its secure storage, as indicated in the Participant Information Sheet.

Personal data will be kept for roughly 6 months, or until it is no longer needed for review. It will be stored electronically on a drive that will be inaccessible to the public and password protected with a password that only I know.

Q21. Is it intended (or possible) that data might be used beyond the present study? (See GN 2.2.16)

YES NO

If YES, please give here an indication of the kind of further use that is intended (or which may be possible).

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If NO, will the data be kept for a set period and then destroyed under secure conditions?

YES NO

If NO, please explain here why not.

Q22. If your study involves work with children and/or vulnerable adults you will require a satisfactory Disclosure and Barring Service (DBS) Disclosure. (See GN 2.2.17) Please indicate as appropriate:

(a) DBS Disclosure not required

(b) DBS Disclosure required and obtained

Please do not embed your DBS Disclosure within the (paper) application documentation. It is recognised that the DBS Disclosure is a confidential document: If a satisfactory DBS Disclosure is required, a copy of this must be forwarded to the ECDA Clerks for review by the Chairman of the relevant ECDA who will note this on a Register which will be kept in a secure place. (Note: only the relevant ECDA Clerk and Chairman will see this document.)

Declaration of Disclosure and Barring Service disclosure

Please state the date on which your most recent DBS Disclosure was obtained …………………….

I declare that I have received NO convictions, cautions, reprimands or final warnings or driving offences resulting in a criminal conviction and that I am not involved with ongoing police investigations since my most recent DBS screening.

I understand that ethics approval is given on the understanding that the three-year validity of my most recent DBS Disclosure will cover the period in which this study will be conducted and that, as a student, I will renew the declaration I make within the School in which I am registered on an annual basis.

I also understand that it is my responsibility to inform the ECDA as soon as a conviction or investigation arise during the period of the study to be conducted.

Enter your name here …………………………………………….. Date …………………………………

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7. REWARDS

Q23.1 Are you receiving any financial or other reward connected with this study? (See UPR RE01, 2.3)

YES NO

If YES, give details here.

Q23.2 Are participants going to receive any financial or other reward connected with the study? (Please note that the University does not allow participants to be given financial inducement (See UPR RE01, 2.3.))

YES NO

If YES, give details here.

Q23.3 Will anybody else (including any other members of the investigative team) receive any financial or other reward connected with this study?

YES NO

If YES, give details here.

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X

X

X

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8. OTHER RELEVANT MATTERS

Q24. Enter here anything else you want to say in support of your application, or which you believe may assist the Committee in reaching its decision.

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APPENDIX 1 – INCREASED HAZARDS AND RISKS This section is to be completed if your answer to Q10 affirms the USE OF INVASIVE PROCEDURES in your study.

Note: You are advised to read GN 2.2.10, 2.2.11, 2.2.12, 2.2.13 & 2.2.18 carefully before you answer the following questions.

QA1. Please give details of the procedures to be used (e.g. injection of a substance, insertion of a catheter, taking of a blood or saliva sample), and any harm, discomfort or distress that their use may cause to participants and/or investigator(s). (See GN 2.2.10)

Consumption of beta alanine that could cause paraesthesia or mild taurine deficiency(won’t occur unless due to severe overdosing and a very unbalanced diet)

Indicate here what precautions will be taken to avoid or minimise any adverse effects.Daily doses will be pre weighed and separated into containers by myself based on weight of participants to assure no overdosing of supplements occurs.

QA2. Will the study involve the administration of any substance(s)? (See GN 2.2.10)

YES NO

If YES,

Give details here of the substance(s), the dose or amount to be given, likely effects (including duration) and any potential hazards to participant(s) and/or investigator(s).

Beta alanine 1.5g to be ingested 4 times per day over a period of 2 weeks, very slight variations calculated based on body weight, for those taking prior to workout they will consume up to 2.5g based on their weight. Potential hazards are only to participants and as aforementioned can be: paraesthesia, taurine deficiency.

Indicate here what precautions will be taken to avoid or minimise any adverse effects.

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QA3. Are there any potential hazards to participant(s) and/or investigator(s) arising from the use of the proposed invasive procedures? (See GN 2.2.10)

YES NO

If YES,

Indicate their nature here.

Indicate here what precautions will be taken to avoid or minimise any adverse effects.

QA4. Will or could the study cause discomfort or distress of a mental or emotional character to participants and/or investigator(s)? (See GN 2.2.11)

YES NO

If YES,

Indicate its nature here

Indicate here what precautions will be taken to avoid or minimise such adverse effects.

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QA5. Medical or other aftercare and/or support must be made available for participants and/or investigator(s) who require it where invasive procedures have been used in the study. Please detail what aftercare and/or support will be available and in what circumstances it is intended to be used. (See UPR RE01, S 2.3 (ii) and GN 2.2.12)

All lab technicians are first aid trained hence they can provide any immediate first aid required, should anything serious happen then the on campus emergency no. will be present as well as 999 and the on campus GP.

QA6.1Please state here previous experience (and/or any relevant training) of the supervisor (or academic member of staff applying for a standard protocol) of investigations involving hazards, risks, discomfort or distress as specified. (See GN 2.2.13)

The supervisor has overseen similar projects before.

QA6.2 Please describe in appropriate detail what you would do should the adverse effects or events which you believe could arise from your study, and which you have mentioned in your replies to the previous questions, occur.

Should anything serious occur on site then first aid can be provided as detailed above and the emergency numbers can be contacted. In terms of serious symptoms from supplementation there is nothing foreseeable.

Please revert to Q15.

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APPENDIX 2 – DOCUMENTS TO BE ATTACHED

Please indicate below which documents are attached to this application:

1 Permission to access groups of participants from student body

2 Permission to use University premises beyond areas of Schools

3 Permission from the off-campus location to be used to carry out this study

4 Risk assessment(s) in respect of hazards/risks affecting both theparticipants and investigator(s) (Q8 and Q11)

5 Copy of Consent Form (See Form EC3 & Form EC4) (Q18)

6 Copy of Form EC6 (Participant Information Sheet) (Q5)

7 Disclosure and Barring Service disclosure

8 A copy of the proposed questionnaire and/or interview schedule (if appropriate for this study). For unstructured methods, please provide details of the subject areas that will be covered and any boundaries that have been agreed with your Supervisor.

9 Any other relevant documents, such as a debrief, meeting report.

EC2:UNIVERSITY OF HERTFORDSHIRE

ETHICS COMMITTEE FOR STUDIES INVOLVING THE USE OF HUMAN PARTICIPANTS(‘ETHICS COMMITTEE’)

FORM EC2: APPLICATION FOR MODIFICATION AND/OR EXTENSION TO AN EXISTING PROTOCOL APPROVAL

1 Protocol Number and Title of original application: (Please provide the original application and attachments. This application will not be accepted without them)

Applicant name: Ernest Didehvar-Sadr

Student registration number: 13035885

Applicant e-mail address: [email protected]

Work address (if appropriate)

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Supervisor’s name: Dr. Fang Lou

Supervisor’s School & Department: School of Life and Medical sciences

Supervisor’s e-mail address: [email protected]

2 Specify the nature of the modification/extension

i Revised title of study (if applicable) The effects of beta alanine on muscular endurance

ii Time (specify months and year): From: 01/03/16 To: 10/04/16

iii Additional worker(s). (Name workers or supervisor to be added to the protocol approval)

Name:

Work address:

E-mail address

iv Change of supervisor from ………………[name] to …………………… [name]Please complete declaration and give reason in Section 3

Declaration by new supervisor:I have reviewed the ethics protocol paperwork for this study and am aware of any conditions which must be adhered to.

Signed ……………………………………………..

v Location of study: H260, G111

vi Details of modification: (Please list all changes being made from the original procedure on a separate sheet) Instead of performing 5 rep maximums for testing the participants will perform 65% of their bodyweight for maximum repetitions for the bench press and 120% of their body weight for maximum repetitions for the deadlift.

3 Reason for extension/modification request(please explain)I wish to change the testing methods for the study as I feel they can be better analysed by using a percentage of the participants’ body weight as a measure for how much to bench press/deadlift and measuring any increases seen by repetitions completed. Additionally, I feel the new testing method will likely increase the chance of any improvements seen as a result of supplementation as it is a better test of anaerobic muscular endurance as trained individuals will likely complete more than 5 reps for those body weight percentages in each exercise. Also, this will rid the obstacle of determining each participants maximum weight for a particular rep range as this can be affected by a whole host of issues making the study less likely to be accurate.

4 Hazards

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Does the modification or extension present additional hazards to the participant/investigator?

YES NO

If YES, a new risk assessment may be necessary. Subject specific forms may also be necessary; you should therefore contact your Supervisor or School to see whether this is the case.

Signature of Applicant ................................................................................Date………………

Support by Supervisor.................................................................................Date………………

EC3:

UNIVERSITY OF HERTFORDSHIREETHICS COMMITTEE FOR STUDIES INVOLVING THE USE OF HUMAN PARTICIPANTS(‘ETHICS COMMITTEE’)

FORM EC3CONSENT FORM FOR STUDIES INVOLVING HUMAN PARTICIPANTS

I, the undersigned [please give your name here, in BLOCK CAPITALS]

……………………………………………………………………………………………………………….…of [please give contact details here, sufficient to enable the investigator to get in touch with you, such as a postal or email address]

…..………………………………………………………………………………………………………………hereby freely agree to take part in the study entitled [The effects of β-Alanine on anaerobic endurance]

…………………………………………………………………………………………………………………..

1 I confirm that I have been given a Participant Information Sheet (a copy of which is attached to this form) giving particulars of the study, including its aim(s), methods and design, the names and contact details of key people and, as appropriate, the risks and potential benefits, and any plans for follow-up studies that might involve further approaches to participants. I have been given details of my involvement in the study. I have been told that in the event of any significant change to the aim(s) or design of the study I will be informed, and asked to renew my consent to participate in it.

2 I have been assured that I may withdraw from the study at any time without disadvantage or having to give a reason.

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3 I have been given information about the risks of my suffering harm or adverse effects. I have been told about the aftercare and support that will be offered to me in the event of this happening, and I have been assured that all such aftercare or support would be provided at no cost to myself.

4 I have been told how information relating to me (data obtained in the course of the study, and data provided by me about myself) will be handled: how it will be kept secure, who will have access to it, and how it will or may be used.

5 I understand that my participation in this study may reveal findings that could indicate that I might require medical advice. In that event, I will be informed and advised to consult my GP. If, during the study, evidence comes to light that I may have a pre-existing medical condition that may put others at risk, I understand that the University will refer me to the appropriate authorities and that I will not be allowed to take any further part in the study.

6 I understand that if there is any revelation of unlawful activity or any indication of non-medical circumstances that would or has put others at risk, the University may refer the matter to the appropriate authorities.

7 I have been told that I may at some time in the future be contacted again in connection with this or another study.

Signature of participant……………………………………..…Date………………………….

Signature of (principal) investigator………………………………………………………Date…………………………

Name of (principal) investigator [in BLOCK CAPITALS please]

…………………………………………………………………………………………………………

EC6: UNIVERSITY OF HERTFORDSHIRE

ETHICS COMMITTEE FOR STUDIES INVOLVING THE USE OF HUMAN PARTICIPANTS(‘ETHICS COMMITTEE’)

FORM EC6: PARTICIPANT INFORMATION SHEET

Title of study

The effects of beta alanine on anaerobic strength as a daily supplement

Introduction

You are being invited to take part in a study. Before you decide whether to do so, it is important that you understand the research that is being done and what your involvement will include. Please take the time to read the following information carefully and discuss it with others if you wish. Do not hesitate to ask us anything that is not clear or for any further information you would like to help you make your decision. Please do take your time to

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decide whether or not you wish to take part. The University’s regulations governing the conduct of studies involving human participants can be accessed via this link:

http://sitem.herts.ac.uk/secreg/upr/RE01.htm

Thank you for reading this.

What is the purpose of this study?

The purpose of this study is to determine whether beta alanine supplementation can increase strength in young males either as a long term daily supplement. The supplementation of beta alanine has been shown to increase muscular endurance and anaerobic running capacity, however it has only been linked with increased anaerobic strength through its use as a stimulant in most pre workout supplements but hasn’t been as of yet been proven to increase anaerobic strength on its own, this will be evaluated as part of the study.

Do I have to take part?

It is completely up to you whether or not you decide to take part in this study. If you do decide to take part you will be given this information sheet to keep and be asked to sign a consent form. Agreeing to join the study does not mean that you have to complete it. You are free to withdraw at any stage without giving a reason. A decision to withdraw at any time, or a decision not to take part at all, will not affect any treatment/care that you may receive (should this be relevant).

Are there any age or other restrictions that may prevent me from participating?

Participants must be male and between the age of 18 and 25 with no underlying medical conditions.

How long will my part in the study take?

If you decide to take part in this study, you will be involved in it for up to 4 weeks.

What will happen to me if I take part?

Potential participants will be prescreened to ensure they aren’t supplementing with anything that could potentially affect the study, such as beta alanine or taurine. In addition to this they will perform a health screen to ensure they have no underlying health conditions, allergies or injuries. In this project, which will be performed in a double blind format, participants will be separated into groups. Groups will be randomly assigned and they will be as follows: group 1 will consist of participants will be given a 2 week supplementation plan for beta alanine that will consist of a daily dose of beta alanine equivalent to 100mg/kg/day spread out across 4 doses a day Group 2 will be given a placebo, 4 times a day for 2 weeks. Following the opening 2 week period each group will test their 5 rep maximums for deadlifts and bench press with the results being privately stored. The study will then be crossed over and each group will swap for 2 weeks and then tested again. In order to ascertain the doses required for each participant they will come in prior to the study to be weighed, and they will also need to perform a baseline for the deadlift and bench press 5 rep maxes, they will also begin taking a food diary so that their diets can be analysed. Prior to consumption they will perform a bench press and deadlift for sets of 5 repetitions at maximum possible weight and then following the supplementation plan provided (either placebo or non-placebo) for 2 weeks will perform the same exercises observing if they are able to lift a heavier weight for the same number of repetitions, form will be monitored and standardised. If participants aren’t dedicated to the supplementation required then they will be removed from the study.What are the possible disadvantages, risks or side effects of taking part?

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The potential risk lies in injuring yourself during the exercises where you will be lifting weights, other than that there will be no risks or disadvantages when performing the exercises. Potential side effects may come from supplementation of the amino acids being studied, in healthy individuals beta alanine can commonly cause mild and harmless paresthesia (sensation of tingling) which usually disappears rapidly, and potentially taurine deficiency in very rare cases.

What are the possible benefits of taking part?

The potential benefits of taking part are most likely to occur due to its role in increased carnosine synthesis, it has been shown to increase muscular endurance and anaerobic running capacity.

How will my taking part in this study be kept confidential?All data collected will be electronically stored and will only be accessible via password, rather than using names each participant will be assigned number that they will be identified with.

What will happen to the data collected within this study?

As earlier stated, the data will be stored electronically behind a password and only accessed by myself for use to complete the study, following completion of the study all data will be destroyed.

Who has reviewed this study?

This study has been reviewed by:

The University of Hertfordshire Health and Human Sciences Ethics Committee with Delegated Authority

The UH protocol number is LMS/UG/UH/02321

Who can I contact if I have any questions?

If you would like further information or would like to discuss any details personally, please get in touch with me, in writing, by phone or by email on 07402050088 (mobile) or at [email protected]

Although we hope it is not the case, if you have any complaints or concerns about any aspect of the way you have been approached or treated during the course of this study, please write to the University’s Secretary and Registrar.

Thank you very much for reading this information and giving consideration to taking part in this study.

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Risk assessment:

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2. Statistical Testing2.1: Bench Press:(1) Baseline, (2) Post-Placebo, (3) Post-β-alanine One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: Repetitions

Source Sig. Partial Eta Squared

Supplement Sphericity Assumed .000 .527

Greenhouse-Geisser .000 .527

Huynh-Feldt .000 .527

Lower-bound .000 .527

Pairwise Comparisons

Measure: Repetitions

(I)

Supplement

(J)

Supplement

Mean

Difference (I-

J) Std. Error Sig.b

1 2 -.091 .421 1.000

3 -2.773* .483 .000

2 1 .091 .421 1.000

3 -2.682* .476 .000

3 1 2.773* .483 .000

2 2.682* .476 .000

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2.2 Deadlift: (1) Baseline, (2) Post-Placebo, (3) Post-β-alanine One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: Repetitions

Source Sig. Partial Eta Squared

Supplement Sphericity Assumed .000 .611

Greenhouse-Geisser .000 .611

Huynh-Feldt .000 .611

Lower-bound .000 .611

Pairwise Comparisons

Measure: Repetitions

(I)

Supplement

(J)

Supplement

Mean

Difference (I-

J) Std. Error Sig.b

1 2 .091 .465 1.000

3 -2.773* .360 .000

2 1 -.091 .465 1.000

3 -2.864* .368 .000

3 1 2.773* .360 .000

2 2.864* .368 .000

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2.3: Deadlift chronologically ordered: (1) Baseline, (2) Test 1, (3) Test 2 One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: Repetitions

Source Sig. Partial Eta Squared

Supplement Sphericity Assumed .043 .139

Greenhouse-Geisser .055 .139

Huynh-Feldt .051 .139

Lower-bound .079 .139

Pairwise Comparisons

Measure: Repetitions

(I)

Supplement

(J)

Supplement

Mean

Difference (I-

J) Std. Error Sig.b

1 2 -1.273* .480 .045

3 -1.409 .557 .058

2 1 1.273* .480 .045

3 -.136 .725 1.000

3 1 1.409 .557 .058

2 .136 .725 1.000

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2.4: Bench Press chronologically ordered: (1) Baseline, (2) Test 1, (3) Test 2 One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: Repetitions

Source Sig. Partial Eta Squared

Supplement Sphericity Assumed .035 .148

Greenhouse-Geisser .048 .148

Huynh-Feldt .045 .148

Lower-bound .070 .148

Pairwise Comparisons

Measure: Repetitions

(I)

Supplement

(J)

Supplement

Mean

Difference (I-

J) Std. Error Sig.b

1 2 -1.091 .465 .086

3 -1.591* .561 .030

2 1 1.091 .465 .086

3 -.500 .747 1.000

3 1 1.591* .561 .030

2 .500 .747 1.000

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2.5: Deadlift 1st and 2nd quartiles: (1) Baseline (2) Post-Placebo (3) Post-β-alanine One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: DLrepetitions

Source

Type III Sum

of Squares df

Mean

Square F Sig.

Partial Eta

Squared

Supplement Sphericity Assumed 32.788 2 16.394 10.505 .001 .512

Greenhouse-

Geisser32.788 1.610 20.360 10.505 .002 .512

Huynh-Feldt 32.788 1.873 17.504 10.505 .001 .512

Lower-bound 32.788 1.000 32.788 10.505 .009 .512

Pairwise Comparisons

Measure: DLrepetitions

(I)

Supplement

(J)

Supplement

Mean

Difference (I-

J) Std. Error Sig.b

95% Confidence Interval for

Differenceb

Lower Bound Upper Bound

1 2 -.364 .650 1.000 -2.231 1.503

3 -2.273* .469 .002 -3.618 -.927

2 1 .364 .650 1.000 -1.503 2.231

3 -1.909* .456 .006 -3.219 -.599

3 1 2.273* .469 .002 .927 3.618

2 1.909* .456 .006 .599 3.219

2.6 Deadlift 3rd and 4th quartiles: (1) Baseline, (2) Post-Placebo, (3) Post-β-alanine One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: DLrepetitions

Source

Type III Sum

of Squares df

Mean

Square F Sig.

Partial Eta

Squared

Supplement Sphericity

Assumed93.818 2 46.909 28.561 .000 .741

Greenhouse-

Geisser93.818 1.577 59.482 28.561 .000 .741

Huynh-Feldt 93.818 1.822 51.480 28.561 .000 .741

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Lower-bound 93.818 1.000 93.818 28.561 .000 .741

Error(Supplemen

t)

Sphericity

Assumed32.848 20 1.642

Greenhouse-

Geisser32.848 15.773 2.083

Huynh-Feldt 32.848 18.224 1.802

Lower-bound 32.848 10.000 3.285

Pairwise Comparisons

Measure: DLrepetitions

(I)

Supplement

(J)

Supplement

Mean

Difference (I-

J)

Std.

Error Sig.b

95% Confidence Interval for

Differenceb

Lower Bound Upper Bound

1 2 .545 .666 1.000 -1.365 2.456

3 -3.273* .524 .000 -4.776 -1.769

2 1 -.545 .666 1.000 -2.456 1.365

3 -3.818* .423 .000 -5.031 -2.606

3 1 3.273* .524 .000 1.769 4.776

2 3.818* .423 .000 2.606 5.031

Based on estimated marginal means

*. The mean difference is significant at the .05 level.

b. Adjustment for multiple comparisons: Bonferroni.

2.7 Bench press 1st and 2nd quartiles: (1) Baseline, (2) Post-Placebo, Post-β-alanine One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: BPrepetitions

Source

Type III Sum

of Squares df

Mean

Square F Sig.

Partial Eta

Squared

Supplement Sphericity Assumed 19.818 2 9.909 3.658 .044 .268

Greenhouse-

Geisser19.818 1.839 10.775 3.658 .049 .268

Huynh-Feldt 19.818 2.000 9.909 3.658 .044 .268

Lower-bound 19.818 1.000 19.818 3.658 .085 .268

Error(Supplement

)

Sphericity Assumed 54.182 20 2.709

Greenhouse-

Geisser

54.182 18.394 2.946

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Huynh-Feldt 54.182 20.000 2.709

Lower-bound 54.182 10.000 5.418

Pairwise Comparisons

Measure: BPrepetitions

(I) Supplement (J) Supplement

Mean Difference

(I-J) Std. Error Sig.a

95% Confidence Interval for

Differencea

Lower Bound Upper Bound

1 2 .182 .644 1.000 -1.667 2.030

3 -1.545 .652 .118 -3.416 .325

2 1 -.182 .644 1.000 -2.030 1.667

3 -1.727 .799 .168 -4.020 .565

3 1 1.545 .652 .118 -.325 3.416

2 1.727 .799 .168 -.565 4.020

Based on estimated marginal means

a. Adjustment for multiple comparisons: Bonferroni.2.8 Bench press 3rd and 4th quartiles:(1) Baseline, (2) Post-Placebo, Post-β-alanine One way RM ANOVA Output

Tests of Within-Subjects Effects

Measure: BPrepetitions

Source

Type III Sum

of Squares df

Mean

Square F Sig.

Partial

Eta

Squared

Supplement Sphericity Assumed 107.636 2 53.818 41.887 .000 .807

Greenhouse-

Geisser107.636 1.653 65.117 41.887 .000 .807

Huynh-Feldt 107.636 1.939 55.519 41.887 .000 .807

Lower-bound 107.636 1.000 107.636 41.887 .000 .807

Error(Supplement

)

Sphericity Assumed 25.697 20 1.285

Greenhouse-

Geisser25.697 16.530 1.555

Huynh-Feldt 25.697 19.387 1.325

Lower-bound 25.697 10.000 2.570

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Pairwise Comparisons

Measure: BPrepetitions

(I) Supplement (J) Supplement

Mean

Difference (I-J) Std. Error Sig.b

95% Confidence Interval for

Differenceb

Lower Bound Upper Bound

1 2 -.364 .560 1.000 -1.972 1.245

3 -4.000* .505 .000 -5.448 -2.552

2 1 .364 .560 1.000 -1.245 1.972

3 -3.636* .364 .000 -4.680 -2.593

3 1 4.000* .505 .000 2.552 5.448

2 3.636* .364 .000 2.593 4.680

Based on estimated marginal means

*. The mean difference is significant at the .05 level.

b. Adjustment for multiple comparisons: Bonferroni.

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3. Sample food diary:A food diary as filled by a participant for the 2nd March, 2016 ("WebMD printable food & fitness journal," 2008).

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4. Repetitions completed

4.1 Deadlift repetitions completed at each state of testing

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Baseline Test 1 Test 21 15 17 122 8 9 73 20 23 174 10 13 125 9 12 116 15 17 157 32 36 388 21 18 249 25 26 30

10 14 11 1311 20 19 2312 23 24 2013 19 17 2114 31 32 3615 9 13 1016 12 11 1317 16 18 1618 14 17 1619 16 15 1720 14 18 1521 12 16 1522 21 22 26

4.2 Bench press repetitions completed at each stage of testing

Baseline Test 1 Test 21 15 16 13

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2 15 17 123 33 37 314 14 16 165 10 10 76 40 43 397 22 20 248 20 23 269 29 32 34

10 18 19 2111 32 30 3512 32 39 3513 31 31 3514 28 28 3115 13 12 1516 24 24 2617 22 23 2118 19 19 2219 27 27 3020 25 28 2521 16 14 1722 27 30 34

4.3 Bench Press Time under tension at each stage of testing

Participant

Baseline Time under tension (s)

Test 1 Time under tension (s)

Test 2 Time under tension (s)

1 35 39 362 34 37 33

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3 84 83 834 43 57 525 27 26 336 92 86 997 49 51 498 50 51 539 78 74 84

10 45 47 5011 80 79 7312 86 88 8313 78 72 7814 66 62 7115 30 35 3216 59 62 6017 47 47 5018 40 41 5119 63 61 6220 60 67 5821 41 47 4322 70 72 69

4.4 Deadlift time under tension at each stage of testing

ParticipantsBaseline Time under

tension (S)Test 1 Time under

tension DL (s)Test 2 Time under

tension DL (s)1 31 38 282 22 18 19

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3 34 36 314 18 20 195 22 22 226 36 38 377 90 92 978 48 46 509 52 54 61

10 29 26 2511 46 46 3712 50 53 4713 51 53 5014 67 66 7415 19 23 2016 27 29 2417 39 43 3318 36 36 3019 35 33 3220 40 44 3321 27 37 2522 59 58 58

5. Health screen template

UNIVERSITY OF HERTFORDSHIRE Researcher:

SCHOOL OF LIFE SCIENCE

HEALTH SCREEN 1

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Title of Study:

Subject Name:

It is important when having volunteered as subject for this study, and having read the briefing sheet for subjects that you answer the following questions. Please do not answer any questions if you consider them intrusive.

1) Do you suffer from high blood pressure, or any heart problems?Yes No

2) Do you often get dizzy, or do you know that you have low blood pressure?Yes No

3) When and what did you last eat?

4) Are you under the influence of alcohol or any other psycho-active substance?Yes No

5) Have you had a cold or flu in the last two weeks?Yes No

6) Are you suffering from any musculo-skeletal injury?Yes No

7) Are you currently taking any medication (over the counter, or prescription)?Yes No

(you do not need to answer “Yes” if you are only taking oral contraceptives, or if you are an asthmatic with an inhaler available)

8) Have you ever been told that you should not exercise?Yes No

9) Do you feel fully fit, and eager to act as subject?

Yes No

Is there any reason, not stated above, why you cannot take part as a subject in this practical?

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Yes No

Signature………………………………………………….. Date:

Checked by (Name): Date:

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