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FACULTY OF SCIENCE,
ENGINEERING AND COMPUTING
School of Life Sciences
BSc (Hons) DEGREE
IN
Sports Science
Hasan Mohammed
K1153242
Effects of BCAA Supplementation on Exercise Capacity
13th
August 2014
Supervisor: Dr. Hannah Moir
WARRANTY STATEMENT
This is a student project. Therefore, neither the student nor Kingston University makes any
warranty, express or implied, as to the accuracy of the data or conclusion of the work
performed in the project and will not be held responsible for any consequences arising out of
any inaccuracies or omissions therein.
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Table of Contents
Acknowledgments.....................................................................................................................3
Abstract.....................................................................................................................................4
1. Introduction..........................................................................................................................5
1.1 BCAA Metabolism...............................................................................................................6
1.2 Central Fatigue.....................................................................................................................8
1.3 Physiological Affects...........................................................................................................9
1.4 Consumption and Dosage...................................................................................................11
1.5 Lactate and Exercise Capacity...........................................................................................12
1.6 Glucose and Exercise Capacity..........................................................................................12
2. Theoretical Framework……………………………………………………………….....14
2.1 Research Question..............................................................................................................14
2.2 Research Aims....................................................................................................................14
2.3 Objectives...........................................................................................................................14
2.4 Operational Definitions......................................................................................................14
3. Methods...............................................................................................................................15
3.1 Literature Review and Study Selection..............................................................................15
3.2 Inclusion/Exclusion Criteria...............................................................................................15
3.3 Data Analysis.....................................................................................................................16
4. Results.................................................................................................................................18
5. Discussion and Conclusions...............................................................................................21
5.1 Limitations.........................................................................................................................22
6. Future Recommendations..................................................................................................23
7. Critical Reflection...............................................................................................................24
References...............................................................................................................................25
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Acknowledgements
May I take this opportunity to show my gratitude to all those who assisted in this work. I am
very grateful for the supervision, guidance and motivation provided throughout this project
by Dr. Hannah Moir. Her invaluable character has provided me with faith in my own
capabilities to carry out such project. She has been approachable and readily available
throughout this project and did not spoon feed all of her knowledge, only to get the best out
of myself and this project for which I thank her for. Her mentoring skills are second to none
and I appreciate all the effort and time she has given.
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Abstract
Branched chain amino acids are becoming more renown in sports drinks as a quick and
efficient supply of energy during exercise. There have been many studies which qualitatively
review this topic but predominantly in relation to central fatigue opposed to peripheral
(metabolic/fatigue) mechanisms. This meta-analysis quantitatively reviews all literature
regarding this topic. Aim: To determine whether BCAA supplementation has an ergogenic
effect on endurance-based exercise capacity. Methods: A search of the literature was carried
out and this that met the inclusion criteria was used for analysis. Six published studies met
the inclusion criteria. The studies included multiple exercise protocols and dosages but either
or both lactate or glucose measures were tested in each study. A total of 9 effect sizes were
established and a total of 67 participants, of which the majority were healthy adult males,
were included in this Meta-analysis. The mean dosage of supplementation across all studies
was 19.45g. Results: This study shows BCAA supplementation improved endurance-based
exercise capacity has; p=0.001 for glucose measures and p=0.000 for lactate measures,
whereby p≤0.05. Conclusion: BCAA supplementation does have an ergogenic effect on
endurance-based exercise capacity thus the null hypothesis can be rejected.
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1. Introduction
Branched-chain-amino-acids (BCAA’s) are known as the umbrella term for three essential
amino acids: leucine, isoleucine and valine. They are known as essential amino acids as they
make up 40% of the daily requirement of all amino acids (Stowers, 2009).BCAA’s cannot be
broken down in the body, rather they are oxidised during exercise (Saris et al., 1989).The
recommended dose for leucine is about 40mg/kg of body weight per day and for isoleucine
and valine approximately 10-30mg/kg per day(Hargreaves, 2006). Although the
recommended daily allowance of BCAA’s were 20% per day, amendments after new
research made the needs of BCAA’s increase to 40% (Stowers, 2009). However this research
was carried out on those who were fasting and living normal lifestyles, not those in need of
more energy or muscle, such as athletes or those carrying out exercise (Zeigler & Filler,
1996). Athletes are now using BCAA’s more commonly as ergogenic aids, thus as RDA
levels have increased, it is likely that BCAA supplementation for athletes should also
increase (Stowers, 2009). During exercise, it has been reported that proteolysis increases and
consequently whole-body-protein and amino acids are used further for energy (De Feo et al.,
2003). As well as the increase in use of BCAA’s during exercise, BCAA plasma
concentrations have shown to be decreased during prolonged exercise, thus supplementation
acts as replenishing this reduction in BCAA availability and consequent energy loss during
prolonged exercise (Bloomstrand et al., 1991). Therefore the protein requirement during
(endurance) exercise is increased and BCAA’s are oxidised further thus need to be
replenished within the diet for increased exercise capacity (Gleeson, 2005).
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1.1 BCAA Metabolism
Once metabolised from protein, most free amino acids are transported to the liver and some
metabolism takes place in the viscera and stomach mucosal areas(Zeigler & Filler, 1996).
However, free BCAA’s are metabolised although metabolised via the liver, they are primarily
oxidised within the muscle and fat (adipose) tissue (Zeigler & Filler, 1996).Some BCAA’s
are exchanged in the intestinal viscera and then travel directly to the bloodstream which
makes them so proficient as an energy source during exercise. Most amino acids can be
broken down in the liver with the exception of BCAA’s. BCAA’s are oxidised from their
converted form called oxo-Keto acids and this essentially means BCAA’s benefits for human
function and exercise capacity is very quick and efficient. They are quick to supply energy
during exercise because they do not have to take time to breakdown in the liver first, opposed
to most other proteins. The enzymes needed to catabolise BCAA’s are known as
mitochondrial dehydrogenase and branched-chain keto acid dehydrogenase (BCKADH).It
has been shown in rats that chronic administration of BCAA’s and a high-protein diet
increases hepatic activity of these enzymes (Shimomura et al., 2000). Keto acids can then be
used by the muscle to resynthesise ATP for energy. However, the product of transiminated
leucine is Alpha-Ketoisocaproic, which can inhibit the breakdown of BCKADH to branched
chain oxo acid (BCOA), an acid that can be used as energy in the liver (Stowers, 2009).Some
supplement companies mistakenly sell the keto acid version of BCAA’s and thus inhibit the
natural breakdown of BCAA’s for energy(Hargreaves, 2006).Once BCAA’s are metabolised
in the liver, the organ muscles or adipose tissue, keto acids can be used to fuel the Krebs
cycle for ATP production and supply energy for all muscles and organs. Furthermore,
BCAA’s can also be converted to glutamine or alinine in the muscle, these amino acids can
then undertake glyconeogenesis within the liver to produce glucose, an essential energy
source during exercise.
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BCAA’s form 35% of muscle tissue and are actively used by the muscle and liver as energy.
Out of all six amino acids, BCAA’s have the most potential as a metabolic energy source for
muscles (Stowers, 2009). Muscle tissue naturally attains 60% of necessary enzymes to
metabolise BCAA’s and it is estimated 3%-18% of energy for all exercise is supplied by
BCAA’s. However, intensity and duration levels can alter this estimate accordingly (Stowers,
2009) and it is often the case that oxidation of BCAA’s succeeds the catabolic capacity
during prolonged endurance exercise (Ohtani et al., 2001).
In essence, leucine is one of the predominant ‘foods’ used as an energy source for muscle
during exercise. The bodies needs for leucine, in the form of BCAA’s, is 25 times greater
than the readily available leucine within the body gained from what is known as the ‘free
amino acid pool’. The body gains this extra need of leucine from supplementation of
BCAA’s, from the free amino pool or breaks down the muscle during workouts for the
luecine necessary for exercise (Zeigler & Filler, 1996).When BCAA’s are taken in the form
of a supplement, the free forms they are taken in surpass the gut and liver and directly enter
the bloodstream. Free forms of BCAA’s are quick to increase blood supply and effect BCAA
circulation for immediate effect on exercise capacity, although this is more prominent when
low levels of glycogen are present (Hargreaves, 2006). Nonetheless, large doses of leucine
are not recommended and Cynober, (2013) states the use of BCAA supplementation should
be in combination with carbohydrates. Two primary ways in which BCAA’s help reduce the
effects of fatigue are its ability to reduce central fatigue via the nervous system and help
produce energy via muscle oxidisation or as a key Krebs cycle component (Cynober, 2013)
and increasing lactate threshold during endurance exercise (Matsumoto et al., 2009).
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1.2Central Fatigue
5-hydroxytryptamine is a serotoninreceptor that is synthesised by the catalyst tryptophan
hydroxylase. As this enzyme is not saturated with substrate, the rate at which 5-HT
synthesises is relevant to the transport of tryptophan across the blood-brain barrier (BBB) as
well as the blood tryptophan concentration within the blood (Young, 1986). The transport of
tryptophan across the BBB is dependent on the amount available for transportation, the
capacity of the BBB transporter, the plasma concentration of tryptophan and the
concentration of other large neutral amino acids (LNAA’s and BCAA’s) which are carried by
the same carrier (Pardridge, 1998). Approximately 10% of total plasma tryptophan is in free
from and the remaining 90% is transported whilst bounded to the protein albumin.
Tryptophan is the only amino acid that binds to albumin. During prolonged endurance
exercise, when free fatty acid levels are elevated in the blood, the level of plasma tryptophan
also increases as free fatty acids and tryptophan compete for the same binding site; albumin
(Curzon et al., 1973). Thus the favourability of the transport of tryptophan into the brain,
when the plasma ratio of tryptophan to free BCAA’s increases, makes the release of 5-HT
from neurons more prominent (Bloomstrand, 2006) and therefore increases central fatigue.
Human studies have shown that during exercise the ratio of plasma tryptophan to BCAA’s
increases and that tryptophan is taken-up by the brain particularly during endurance exercise,
possibly increasing the synthesis of 5-hydroxytryptamine (5-HT), a serotonin receptor.
Once tryptophan enters the brain it causes the brain to release serotonin, an important
hormone that produces fatigue and tiredness (Cynober, 2013). Theory suggests the ingestion
of BCAA’s increases tryptophan/5-HT’s metabolism, which may reduce its uptake within the
brain and thus delay fatigue. A study by Bloomstrand et al., (1997) showed that when
BCAA’s supplied to humans during a standardised cycle ergometer exercise, their ratings of
perceived exertion (RPE) and consequent central fatigue reduced. Cognitive test
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performances also improved after a competitive 30km cross-country race, suggesting
enhanced brain activity which may relate to less fatigue. Furthermore Mittleman et al., (1998)
suggested that central fatigue is increased in heat and thus BCAA’s effectiveness in delaying
fatigue also increases. In this study physical performance measured by, the time to exhaustion
during cycling, improved by approximately 16 minutes for men and women during endurance
exercise at 40% VO2max in the heat. Subjects were given 5ml-kg-1 body weight of a solution
containing either 5.88 g-L-1 of polydextrose as the placebo or 5.88 g-L-1 of BCAA’s every 30
minutes during exercise. However, another similar study by Watson et al., (2004) where male
participants cycled to volitional exhaustion at 50% VO2max in a warm environment consumed
four aliquots of 250ml (3g) of a 12g/L-1 BCAA or placebo solution 30mins prior to exercise
and 150ml every 15min during exercise. The participants had shown no effect in the delay of
fatigue; placebo – 26.9min opposed to BCAA – 29.2min. Thus causing indefinite effects of
BCAA supplementation and exercise capacity.
1.3 Physiological Affects
The effects BCAA’s have on energy is approximately 3-18% of total energy. The amino
acids; isoleucine, leucine and valine can also act as key components of the Krebs cycle to
supply energy, making the three components of BCAA’s a valuable energy source (Cynober,
2013).The uses of BCAA’s have been shown to be effective predominantly with endurance-
based activities and in terms of its physiological effects; one month’s oral supplementation of
9 essential amino acids, including BCAA’s, had increased fasting glucose and decreased
creatine phosphokinase (a key enzyme which breaks down creatine and to resynthesise ATP
and supply energy via the predominantly anaerobic ATP-PC energy system)activity in
middle-long distance runners. Taken before and after workouts, marathoners as well as
cyclists have shown positive effects during and before events where improvements in
cognitive performance and reductions in time have been shown. Reductions in lactate, a key
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component of exercise capacity and muscle mass loss has also shown to be reduced (Ohtani
et al., 2001).
BCAA’s enter the Krebs cycle directly as acetyl-CoA and not via the glycolytic pathway
whereby pyruvate is converted to lactate by lactate dehydrogenase, a wasteful by-product that
may inhibit exercise capacity (Harper et al., 1984). A study by Matsumoto et al., (2009)
shows BCAA metabolism does not produce lactate and is considered to decrease lactate
levels after supplementation. As lactate is a by-product of glycolysis during energy
metabolism, ones lactate threshold and lactate concentration are considered as predictors of
endurance exercise capacity (Yoshida et al., 1987). A study carried out by De Palo et al.,
(2001) reported lactate levels are suppressed within the blood during exercise following the
chronic supplementation of BCAA’s. The study led by De Palo et al., (2001) had given
triathletes 30 days of chronic BCAA supplementation (0.2g/kg-1) and 9.64g of BCAA oral-
supplementation before exercise consisting of 60mins at 70% VO2max.Furthermore,a study
carried out by Matsumoto et al., (2009) proved that 6 day supplementation of a BCAA drink
increased lactate threshold by increasing workload levels at LT as well as VO2maxagainst a
placebo. The study exercise test was an incremental loading exercise until exhaustion using a
cycle ergometer; the test drink amounted to 500ml and was given 15min prior exercise.
Participants took 1500ml/d during the 6 prior days to testing and it was concluded BCAA
ingestion before exercise increases BCAA supply as an energy source to the muscle during
endurance exercise. Thus it was theorised that the increase of acetyl-CoA to the Krebs cycle,
via the BCAA catabolic pathway, inactivates the glycolytic pathway and consequently
suppresses lactate production during the test. This suggests that a reduction in carbohydrate
breakdown and lactate production increases endurance exercise capacity by inactivating the
glycolytic energy system.
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1.4 Consumption and Dosage
On average, BCAA’s make up about 15% of total amino acid content within food protein
(Gleeson, 2005). A Tour De France cyclist averages 25 Mj/d over a 2-3week period (Saris et
al., 1989). Although protein in a tour de Frances’ diet may be relatively less, much energy is
consumed in the form of carbohydrates, the protein is about 12% of total body energy for a
tour de France cyclist. The elite cyclist consumes about 3000 kJ as protein thus about 19g of
this protein are BCAA’s (Saris et al., 1989).
Some studies have compared combined and separated glucose and BCAA supplementation
(Madsen et al., 1996; Calders et al., 1999) and formulas are found to have both ergogenic aids
within. The recommended dosage of BCAA’s is 3-20g a day, before and after workouts. A
study carried out on 23 rugby players, who carried out intensive training, identifying the
effects of a mixture of 9 essential amino acids, including leucine, isoleucine, valine and
carbohydrates, showed significant improvements in vigour and earlier recovery from fatigue
after 90 days of supplementation. They were given 3.6g twice, daily, for 90 days. This
amount has been credited by most studies indicating 7-12g during endurance events, mixed
into carbohydrate solution (Bloomstrand & Newsholme, 1992; Davis et al., 1999; Mero,
1999; Koba et al., 2007). A study by Tipton et al., (2004) showed whey protein ingestion
resulted in a higher amount of blood BCAA concentrations, suggesting an increase in
absorption when taken with whey protein or carbohydrates. Thus this study specifically looks
at post lactate and glucose concentrations after BCAA supplementation as an indicator of
endurance exercise capacity. However, dosage amount may critically vary amongst
individuals which studies did not account for.
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1.5 Lactate and Exercise Capacity
The blood lactate curve and lactate thresholds have recently become an important factor in
the assessment of exercise capacity (Faude et al., 2009). Lactate threshold, inversely related
to blood lactate concentrations (bLa), has been always used as an index of endurance exercise
capacity. In exercise intolerant mice with disrupted branched chain amino acid metabolism,
increased rates of lactate release from skeletal muscle during exercise were described (She et
al., 2010). It is a trend with many studies that graded incremental tests eliciting a rise in blood
lactate concentration have been used to determine lactate thresholds or curves which indicate
exercise capacity (Faude et al., 2009). During the first half of the 20th century VO2 max levels
were the most common means of evaluating endurance capacity (Faude et al., 2009) and was
developed by Hill et al., (1923). However, in the 1960’s the method by which lactate
concentrations were measured was by capillary blood samples, which led to an increased
popularity of bLa to assess endurance capacity (Hollmann, 2001) and has now become the
most influential factor in the diagnosis of endurance performance/capacity in sport (Jones,
2006).Therefore this study uses bLa as a means of assessing exercise capacity and the
improvements it may have due to BCAA supplementation, as it is generally accepted the
lower the bLa concentration at a given workload the better ones endurance (exercise)
capacity (Yoshida et al., 1990; Bosquet et al., 2002).
1.6Glucose and Exercise Capacity
It has been outlined by Peronnet & Thibault, (1989) that the physiological basis of endurance
capacity, determined by aerobic endurance, is not clearly grasped. It is, however, a
combination of several factors one of which includes the capacity to spare carbohydrates in
the form of glucose. The capacity to save carbohydrate as a reserve fuel by using more fatty
acids as energy substrates increases ones (endurance) exercise capacity (Foster et al.,
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1978).Thus this study uses the means of blood glucose levels (bGl) as a diagnosis of
endurance-based exercise capacity. However, overall bLa are known to be influenced by
depleted glycogen stores (proceeding exhaustive/endurance exercise) (Reilly & Woodbridge,
1999). For example, low bLa at the same work rates have been shown in glycogen-depleted
subjects compared with a subject in normal condition. This may lead to a lower bLa and
should not be interpreted as an increased exercise capacity (Maassen & Busse, 1989).
Although studies suggest BCAA supplementation positively increases endurance exercise
capacity, other studies have shown no effect as mentioned above by Watson et al., (2004) and
as a result, disagreements remain with the effectiveness of BCAA ingestion on endurance
exercise (Matsumoto et al., 2009). However, these studies used different timing and dosages
thus affecting the comparison in relation to the effects of acute BCAA supplementation.
Many studies have identified BCAA’s effects on central fatigue mechanisms opposed to
peripheral fatigue mechanisms such as metabolism or fatigue. As no (Meta) analysis has been
carried out to determine whether BCAA supplementation has an ergogenic effect on exercise
capacity. This study looks at the effects BCAA supplementation has on metabolic (bGl) and
fatigue (bLa) mechanisms in relation to exercise capacity.
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2. Theoretical Framework
2.1 Research Questions
Does BCAA supplementation have an effect on exercise capacity?
2.2 Research Aims
To determine whether BCAA supplementation has an ergogenic effect on endurance-based
exercise capacity.
2.3 Objectives
To quantitatively review the research of BCAA supplementation and its effects on exercise
capacity.
H1- BCAA supplementation will enhance exercise capacity.
H0 - BCAA supplementation will have no ergogenic effect on exercise capacity.
2.4 Operational Definitions
Endurance exercise defined as any exercise mode in excess of 60 minutes work
Work defined as an increase in HR due to physical activity
Exercise capacity is defined as work until exhaustion measured by lactate or glucose
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3. Methods
3.1 Literature Review and Study Selection
The literature used within this meta-analysis was searched using the databases of
GoogleScholar, SPORTDiscuss and PUBMED in July 2014. A variety of key search words
and terms were used, these included but were not limited to: ‘BCAA’, ‘Branched-Chain-
Amino-Acids’, ‘supplementation’, ‘exercise’, ‘capacity’, ‘endurance’, ‘lactate’, ‘glucose’.
Once all potential studies had been identified, studies were included/excluded for analysis in
accordance with the inclusion/exclusion criteria outlined below. Characteristics included
relevant exercise modes such as endurance exercise and lactate/glucose measures.
Homogeneity of participants was considered but inclusion criteria remained as outlined.
3.2 Inclusion/Exclusion Criteria
This meta-analysis was limited to journal articles using human participants in a placebo or
controlled double-blinded condition. These articles were published in English peer-reviewed
journals which had a BCAA supplementation group, possibly including a carbohydrate
solution and a placebo group (PLA). A summary of the processes used to select relevant
articles to be used in the current meta-analysis are outlined in Fig. 1. Studies that were not
included in the bLa analysis had no relevant info (Matsumoto et al., 2009; Greer et al., 2011;
Pasiakos et al., 2011). Other studies that were not included had an inappropriate testing
strategy (MacLean et al., 1996, where an anaerobic-based dynamic knee extensor exercise
was used as a dependant variable), those with inappropriate experimental groups such as
using rats or mice (Shimomura et al., 2000), or no specific numerical data in a table to use for
analysis (Madsen et al., 1996; Coombes et al., 2000; Shimomura et al., 2004; Koba et al.,
2007), or those that only seem to be available as abstracts (Varnier et al., 1994; Gualano et
16
al., 2011) and one that was only available in a Chinese journal hence was not accessible
(Jianhong & Zhihong, 2005).
3.3 Data Analysis
Pre- and Post-supplementation mean and standard-deviations (SD) were obtained from the
original data from the studies used to establish effect size (ES).The effectiveness of the
supplement was quantified by establishing the effects size(Δ) which was calculated by the
difference between the post and pre-supplementation measures divided by the SD of the
control group (Gene Glass’s approach). The control’s group standard deviation is used as it is
not affected by the treatment (Glass et al., 1981).A forest plot as well as 95% confidence
intervals (p≤0.05) displaying the findings of BCAA supplementation to be effective or not
(on exercise capacity) was created by inputting raw data into Comprehensive Meta Analysis
V2 software. Where shown in Fig. 1, n=number of study’s. Two out of six studies did not
provide data indicating clear numerical lactate measurements, these data were instead given
in a figure which was difficult to use for accurate lactate measures to use in this analysis,
hence were excluded. Where information was not available, ‘no relevant data’ was stated, see
Table 1.
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Fig. 1. Flow chart showing the systematic review of studies and selection strategy
Studiesthatwere excludeddue
to irrelevantsample,predictors
and outcomes(n=16)
Studiesusedforfurtherevaluation
(n=10)
Total studies excluded(n=5)
No relevantavailabledatafor
lactate (n=2)
Studiesincludedincurrent
meta-analysis(n=5)
Potential relevantstudies
identified(n=26)
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4. Results
Table 1.Effects of BCAA supplementation on glucose (G) and lactate (L) levels on
endurance-based exercise capacity.
Study Sample Exercise Protocol Supplementation
Protocol
Total
Dosage of
BCAA (g)
Effect Size
Cheuvront et
al., (2004)
Watson et al.,
(2004)
Matsumoto et
al., (2009)
7 Healthy
Males
8 Males
21 Trained
Males
60min cycling @
50% VO2 max.
Then,
30min time-trial
Cycling to volitional
exhaustion @ 50%
VO2 max.
Incremental loading
exercise test with
cycle ergometer until
exhaustion
60g/l glucose,
10g/l BCAA.
Total of 1.4L;
200ml at 15min
intervals
4 250ml aliquots
of a 12g/l BCAA
solution every
30min prior
exercise for a total
of 120min
150ml every
15min throughout
exercise
6ml BCAA,60ml
Carbohydrate
solution;
1500ml/d for 6d
500ml 15min
prior exercise test
24.4
54
2
(3.23-0.9)/1.27
ES = 1.83 (L)
(6.15-5.02)/1.1
ES = 1 (G)
(1.53-
0.59)/0.23
ES = 4.1 (L)
(4.1-4)/1
ES = 0.1 (G)
(114-101)/15
ES = 0.86(G)
No Relevant
info for Lactate
Hsu et al.,
(2011)
14 Healthy
Males
5min warm-up @
55% VO2 max on a
treadmill
Then, ran @ 75%
VO2 max for 30min.
Thereafter,intensity
increased by 1%
until exhaustion
200ml of BCAA
drink; valine
(0.5g), leucine
(1.0g), isoleucine
(0.5g) and
carbohydrate
(12.1g) consumed
post-exercise
2 (3.6-1)/0.4
ES = 6.5 (L)
(80.3-78.1)/1.5
ES = 1.47 (G)
19
Study Sample Exercise Protocol Supplementation
Protocol
Total
Dosage of
BCAA (g)
Effect Size
Greer et al.,
(2011)
Pasiakos et al.,
(2011)
9 Untrained
Males
8 Adults
3 90min Cycling
bouts @ 55% VO2
max
Cycle ergometer @
60% VO2 max
BCAA drink;
valine (7.3g),
leucine (12.2g),
isoleucine (4.8g).
Administered
5min prior
exercise and at
60min during
exercise bout
125ml of 10g
BCAA drink
(leucine enriched)
throughout
exercise
24.3
10
(102.36-
102.34)/18.84
ES = 0.001 (G)
No relevant
info for Lactate
(4.9-4.7)/0.1
ES = 2 (G)
No relevant
info for Lactate
20
Study name Statistics for eachstudy Stddiffinmeans and95%CI
Stddiff Standard Lower Upper
inmeans error Variance limit limit Z-Value p-Value
Cheuvrontetal., (2004) 0.576 0.545 0.298 -0.493 1.645 1.056 0.291
Watsonetal., (2004) 0.050 0.500 0.250 -0.930 1.030 0.100 0.920
Matsumotoetal., (2009) 1.036 0.329 0.108 0.392 1.680 3.152 0.002
Hsuetal., (2011) 0.802 0.393 0.154 0.032 1.572 2.042 0.041
Greeretal., (2011) 0.002 0.471 0.222 -0.922 0.926 0.005 0.996
Pasiakos etal., (2011) 0.316 0.503 0.253 -0.670 1.302 0.629 0.530
0.576 0.178 0.032 0.227 0.925 3.237 0.001
-1.00 -0.50 0.00 0.50 1.00
Treatment effect p<0.05
Fig 2. Statistical findings of BCAA supplementation on Glucosemeasures Evaluation copy
Studyname Statisticsfor each study Std diffin meansand 95% CI
Std diff Standard Lower Upper
in means error Variance limit limit Z-Value p-Value
Cheuvront et al., (2004) 1.105 0.574 0.329 -0.020 2.230 1.926 0.054
Watson et al., (2004) 0.980 0.529 0.280 -0.057 2.017 1.852 0.064
Hsu et al., (2011) 1.554 0.431 0.186 0.709 2.399 3.603 0.000
1.269 0.289 0.083 0.703 1.835 4.394 0.000
-1.00 -0.50 0.00 0.50 1.00
Treatment effect p<0.05
Fig 3. Statistical findings of BCAA supplementation on Lactate measures
Evaluation copy
21
5. Discussion and Conclusions
This meta-analysis shows the overall effect of BCAA supplementation on endurance-based
exercise capacity has a significant effect where p≤0.05; p=0.001 for glucose measures and
p=0.000 for lactate measures (see fig. 2. and fig.3.). The analysis indicates that a dosage of
19.45g (mean dosage across all studies) of BCAA supplementation will have a positive effect
on lower bLa and higher bGl indicating an enhanced exercise capacity for endurance-based
activities. The results of this study are in agreement with previous studies including (Hsu et
al., 2011; Matsumoto et al., 2011). In accordance to conventional effect size values, (Cohen,
1962) glucose may consider a ‘medium’ effect size for BCAA supplementation to be
applicable and lactate may be considered to have a ‘large’ effect size; (d=0.032, d=0.083
respectively).
The results for this study may be surprising as only two of six studies show significant
differences between treatment and control groups in relation to lactate (Hsu et al., 2011;
Motsumoto et al., 2011). However, the glucose trials identify significant differences which
may be the reason for the significance of this study. It is important that these measurements
are used together to determining the effects of BCAA supplementation as Cynober, (2013)
states the use of BCAA’s should be in combination with carbohydrates. A study by Hsu et
al., (2011) showed a BCAA drink containing 24.2g of carbohydrates may have enhanced the
insulin response during post-exercise period. This suggests when BCAA and carbohydrates
are mixed into a solution; it enhances anabolic responses during recovery (Hsu et al., 2011).
Hsu et al., (2011) also found BCAA drink did not stimulate an increase in bLa, which is
consistent with this study. The reason metabolism (of glucose) may be significant is the fact
that BCAA’s may act as a substrate for muscle metabolism (Hsu et al., 2011).
Amino acids have recently become more prominent in sport beverages due to an apparent
reduction on muscle damage when consumed before or after exercise (Greer et al., 2007).
22
However, a concern arises for adding BCAA’s to spots drinks as it may add a potential
carbon drain on the Krebs cycle. In the process of transimination, the first stop of BCAA
catabolism, leucine is accepted and forms glutamate. When pyruvate (which forms lactate) is
not available to supple alinine aminotransferase reaction, it may lead to a depletion of the
Krebs cycle flux to produce ATP, during the oxidation of leucine, an essential amino acid
(Wagenmakers, 1999).
In conclusion the metabolic (bGl) and fatigue (bLa) mechanisms used as diagnosing exercise
capacity has shown to be effective in enhancing exercise capacity by BCAA
supplementation. Therefore the null hypothesis can be rejected in accordance with the above
outlined glucose and lactate significance levels.
5.1 Limitations
The results from this study do go against some previous studies such as (Watson et al., 2004;
Cheuvont et al., 2004) but these studies were done less recently and thus may be irrelevant
when compared with those that have been done more recently. Nonetheless, the studies
included in this meta-analysis may show not significant measures compared with a placebo
due to a better understanding of how to perform trials or a possible ‘placebo effect’ (Laursen
et al., 2003).
This study uses incremental exercise to exhaustion and exercise lasting 60mins or more to
elicit rise in bLa. However, this measure of endurance capacity is hindered by many factors
that may influence overall lactate levels. The magnitude of endurance gains is difficult to
predict by using bLa (Faude et al., 2009) and may lead to this study to lack internal validity.
Nevertheless, studies have shown bLa to be a renowned measurement in the diagnoses of
exercise capacity (Yoshida et al., 1990; Bosquet et al., 2002; Faude et al., 2009).
23
6. Future recommendations
In terms of the consumption of BCAA’s, they are safe within recommended dosages and
have shown to improve exercise capacity within this meta-analysis if an average of 19.45g is
taken according to the protocols used in this study. The use of amino acid supplements is not
prohibited by the World-Anti-Doping Agency (WADA) and thus makes it a legal substance
(Williams, 2005). This study proves the increased effects of endurance-based exercise
capacity due to BCAA supplementation and thus may be an effective ergogenic aid for
marathoners, cyclists and those involved in endurance activities. Coaches may use such
supplements to quickly supply energy to athletes for endurance exercise as mentioned,
BCAA’s predominantly metabolise in the muscle and fat tissue (Zeigler & Filler, 1996).
However, the majority of participants in this study were healthy male adults and not elite
athletes. The results in this meta-analysis were also calculated using comparatively few
studies (n=6) which may make this study lack ecological validity. Furthermore, this study
does not consider time-trial based results for measurements and although stated, does not take
dosage effects into calculation. Findings of BCAA supplementation remain equivocal in
particular its physiological effects on exercise capacity. Bloomstrand, (2001) reported
reductions on perceived exertion ratings (RPE) and consequent central fatigue after BCAA
supplementation during prolonged exercise, suggesting improvements in physical
performance but the named study used central fatigue mechanisms to identify exercise
enhancements. This highlights the need for future studies investigating the effects of BCAA
supplementation on exercise capacity, in relation to peripheral fatigue mechanisms, is
necessary with dosage, time-trial (performance), different mode(s) of exercise (muscular
strength/power output) and different types as well as homogeneity amongst subjects as
factors taken into consideration.
24
7. Critical reflection
To undertake this research key life skills and technical skills had to be applied in order to
successfully and efficiently carry out work. In doing so, time management skills to stick to
deadlines had to be adhered to as well as excellent communication skills, which included
emails to the supervisor, were made throughout this research project. Discipline to carry out
work out of university hours was implied to keep up-to-date. The organisation of work was
parallel to a successful project; organising relevant and irrelevant research papers into folders
for future reference and usage within this analysis was very important to stay focused and
precise in this project.
Key transferable skills were also learnt and utilised such as the use of meta-analysis software
(Comprehensive Meta Analysis V2), Microsoft Word, Outlook and PowerPoint for a high
level of presentation of data, sending and constantly being aware of emails and presenting
work in a slide show using PowerPoint.
To be able to carry out this research project alongside other commitments such as a job has
increased my ability to juggle many works at a single time and not only plan, but effectively
manage time according to deadlines.
25
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NEW FINAL Effects of BCAA's on Exercise Capacity

  • 1. 1 FACULTY OF SCIENCE, ENGINEERING AND COMPUTING School of Life Sciences BSc (Hons) DEGREE IN Sports Science Hasan Mohammed K1153242 Effects of BCAA Supplementation on Exercise Capacity 13th August 2014 Supervisor: Dr. Hannah Moir WARRANTY STATEMENT This is a student project. Therefore, neither the student nor Kingston University makes any warranty, express or implied, as to the accuracy of the data or conclusion of the work performed in the project and will not be held responsible for any consequences arising out of any inaccuracies or omissions therein.
  • 2. 2 Table of Contents Acknowledgments.....................................................................................................................3 Abstract.....................................................................................................................................4 1. Introduction..........................................................................................................................5 1.1 BCAA Metabolism...............................................................................................................6 1.2 Central Fatigue.....................................................................................................................8 1.3 Physiological Affects...........................................................................................................9 1.4 Consumption and Dosage...................................................................................................11 1.5 Lactate and Exercise Capacity...........................................................................................12 1.6 Glucose and Exercise Capacity..........................................................................................12 2. Theoretical Framework……………………………………………………………….....14 2.1 Research Question..............................................................................................................14 2.2 Research Aims....................................................................................................................14 2.3 Objectives...........................................................................................................................14 2.4 Operational Definitions......................................................................................................14 3. Methods...............................................................................................................................15 3.1 Literature Review and Study Selection..............................................................................15 3.2 Inclusion/Exclusion Criteria...............................................................................................15 3.3 Data Analysis.....................................................................................................................16 4. Results.................................................................................................................................18 5. Discussion and Conclusions...............................................................................................21 5.1 Limitations.........................................................................................................................22 6. Future Recommendations..................................................................................................23 7. Critical Reflection...............................................................................................................24 References...............................................................................................................................25
  • 3. 3 Acknowledgements May I take this opportunity to show my gratitude to all those who assisted in this work. I am very grateful for the supervision, guidance and motivation provided throughout this project by Dr. Hannah Moir. Her invaluable character has provided me with faith in my own capabilities to carry out such project. She has been approachable and readily available throughout this project and did not spoon feed all of her knowledge, only to get the best out of myself and this project for which I thank her for. Her mentoring skills are second to none and I appreciate all the effort and time she has given.
  • 4. 4 Abstract Branched chain amino acids are becoming more renown in sports drinks as a quick and efficient supply of energy during exercise. There have been many studies which qualitatively review this topic but predominantly in relation to central fatigue opposed to peripheral (metabolic/fatigue) mechanisms. This meta-analysis quantitatively reviews all literature regarding this topic. Aim: To determine whether BCAA supplementation has an ergogenic effect on endurance-based exercise capacity. Methods: A search of the literature was carried out and this that met the inclusion criteria was used for analysis. Six published studies met the inclusion criteria. The studies included multiple exercise protocols and dosages but either or both lactate or glucose measures were tested in each study. A total of 9 effect sizes were established and a total of 67 participants, of which the majority were healthy adult males, were included in this Meta-analysis. The mean dosage of supplementation across all studies was 19.45g. Results: This study shows BCAA supplementation improved endurance-based exercise capacity has; p=0.001 for glucose measures and p=0.000 for lactate measures, whereby p≤0.05. Conclusion: BCAA supplementation does have an ergogenic effect on endurance-based exercise capacity thus the null hypothesis can be rejected.
  • 5. 5 1. Introduction Branched-chain-amino-acids (BCAA’s) are known as the umbrella term for three essential amino acids: leucine, isoleucine and valine. They are known as essential amino acids as they make up 40% of the daily requirement of all amino acids (Stowers, 2009).BCAA’s cannot be broken down in the body, rather they are oxidised during exercise (Saris et al., 1989).The recommended dose for leucine is about 40mg/kg of body weight per day and for isoleucine and valine approximately 10-30mg/kg per day(Hargreaves, 2006). Although the recommended daily allowance of BCAA’s were 20% per day, amendments after new research made the needs of BCAA’s increase to 40% (Stowers, 2009). However this research was carried out on those who were fasting and living normal lifestyles, not those in need of more energy or muscle, such as athletes or those carrying out exercise (Zeigler & Filler, 1996). Athletes are now using BCAA’s more commonly as ergogenic aids, thus as RDA levels have increased, it is likely that BCAA supplementation for athletes should also increase (Stowers, 2009). During exercise, it has been reported that proteolysis increases and consequently whole-body-protein and amino acids are used further for energy (De Feo et al., 2003). As well as the increase in use of BCAA’s during exercise, BCAA plasma concentrations have shown to be decreased during prolonged exercise, thus supplementation acts as replenishing this reduction in BCAA availability and consequent energy loss during prolonged exercise (Bloomstrand et al., 1991). Therefore the protein requirement during (endurance) exercise is increased and BCAA’s are oxidised further thus need to be replenished within the diet for increased exercise capacity (Gleeson, 2005).
  • 6. 6 1.1 BCAA Metabolism Once metabolised from protein, most free amino acids are transported to the liver and some metabolism takes place in the viscera and stomach mucosal areas(Zeigler & Filler, 1996). However, free BCAA’s are metabolised although metabolised via the liver, they are primarily oxidised within the muscle and fat (adipose) tissue (Zeigler & Filler, 1996).Some BCAA’s are exchanged in the intestinal viscera and then travel directly to the bloodstream which makes them so proficient as an energy source during exercise. Most amino acids can be broken down in the liver with the exception of BCAA’s. BCAA’s are oxidised from their converted form called oxo-Keto acids and this essentially means BCAA’s benefits for human function and exercise capacity is very quick and efficient. They are quick to supply energy during exercise because they do not have to take time to breakdown in the liver first, opposed to most other proteins. The enzymes needed to catabolise BCAA’s are known as mitochondrial dehydrogenase and branched-chain keto acid dehydrogenase (BCKADH).It has been shown in rats that chronic administration of BCAA’s and a high-protein diet increases hepatic activity of these enzymes (Shimomura et al., 2000). Keto acids can then be used by the muscle to resynthesise ATP for energy. However, the product of transiminated leucine is Alpha-Ketoisocaproic, which can inhibit the breakdown of BCKADH to branched chain oxo acid (BCOA), an acid that can be used as energy in the liver (Stowers, 2009).Some supplement companies mistakenly sell the keto acid version of BCAA’s and thus inhibit the natural breakdown of BCAA’s for energy(Hargreaves, 2006).Once BCAA’s are metabolised in the liver, the organ muscles or adipose tissue, keto acids can be used to fuel the Krebs cycle for ATP production and supply energy for all muscles and organs. Furthermore, BCAA’s can also be converted to glutamine or alinine in the muscle, these amino acids can then undertake glyconeogenesis within the liver to produce glucose, an essential energy source during exercise.
  • 7. 7 BCAA’s form 35% of muscle tissue and are actively used by the muscle and liver as energy. Out of all six amino acids, BCAA’s have the most potential as a metabolic energy source for muscles (Stowers, 2009). Muscle tissue naturally attains 60% of necessary enzymes to metabolise BCAA’s and it is estimated 3%-18% of energy for all exercise is supplied by BCAA’s. However, intensity and duration levels can alter this estimate accordingly (Stowers, 2009) and it is often the case that oxidation of BCAA’s succeeds the catabolic capacity during prolonged endurance exercise (Ohtani et al., 2001). In essence, leucine is one of the predominant ‘foods’ used as an energy source for muscle during exercise. The bodies needs for leucine, in the form of BCAA’s, is 25 times greater than the readily available leucine within the body gained from what is known as the ‘free amino acid pool’. The body gains this extra need of leucine from supplementation of BCAA’s, from the free amino pool or breaks down the muscle during workouts for the luecine necessary for exercise (Zeigler & Filler, 1996).When BCAA’s are taken in the form of a supplement, the free forms they are taken in surpass the gut and liver and directly enter the bloodstream. Free forms of BCAA’s are quick to increase blood supply and effect BCAA circulation for immediate effect on exercise capacity, although this is more prominent when low levels of glycogen are present (Hargreaves, 2006). Nonetheless, large doses of leucine are not recommended and Cynober, (2013) states the use of BCAA supplementation should be in combination with carbohydrates. Two primary ways in which BCAA’s help reduce the effects of fatigue are its ability to reduce central fatigue via the nervous system and help produce energy via muscle oxidisation or as a key Krebs cycle component (Cynober, 2013) and increasing lactate threshold during endurance exercise (Matsumoto et al., 2009).
  • 8. 8 1.2Central Fatigue 5-hydroxytryptamine is a serotoninreceptor that is synthesised by the catalyst tryptophan hydroxylase. As this enzyme is not saturated with substrate, the rate at which 5-HT synthesises is relevant to the transport of tryptophan across the blood-brain barrier (BBB) as well as the blood tryptophan concentration within the blood (Young, 1986). The transport of tryptophan across the BBB is dependent on the amount available for transportation, the capacity of the BBB transporter, the plasma concentration of tryptophan and the concentration of other large neutral amino acids (LNAA’s and BCAA’s) which are carried by the same carrier (Pardridge, 1998). Approximately 10% of total plasma tryptophan is in free from and the remaining 90% is transported whilst bounded to the protein albumin. Tryptophan is the only amino acid that binds to albumin. During prolonged endurance exercise, when free fatty acid levels are elevated in the blood, the level of plasma tryptophan also increases as free fatty acids and tryptophan compete for the same binding site; albumin (Curzon et al., 1973). Thus the favourability of the transport of tryptophan into the brain, when the plasma ratio of tryptophan to free BCAA’s increases, makes the release of 5-HT from neurons more prominent (Bloomstrand, 2006) and therefore increases central fatigue. Human studies have shown that during exercise the ratio of plasma tryptophan to BCAA’s increases and that tryptophan is taken-up by the brain particularly during endurance exercise, possibly increasing the synthesis of 5-hydroxytryptamine (5-HT), a serotonin receptor. Once tryptophan enters the brain it causes the brain to release serotonin, an important hormone that produces fatigue and tiredness (Cynober, 2013). Theory suggests the ingestion of BCAA’s increases tryptophan/5-HT’s metabolism, which may reduce its uptake within the brain and thus delay fatigue. A study by Bloomstrand et al., (1997) showed that when BCAA’s supplied to humans during a standardised cycle ergometer exercise, their ratings of perceived exertion (RPE) and consequent central fatigue reduced. Cognitive test
  • 9. 9 performances also improved after a competitive 30km cross-country race, suggesting enhanced brain activity which may relate to less fatigue. Furthermore Mittleman et al., (1998) suggested that central fatigue is increased in heat and thus BCAA’s effectiveness in delaying fatigue also increases. In this study physical performance measured by, the time to exhaustion during cycling, improved by approximately 16 minutes for men and women during endurance exercise at 40% VO2max in the heat. Subjects were given 5ml-kg-1 body weight of a solution containing either 5.88 g-L-1 of polydextrose as the placebo or 5.88 g-L-1 of BCAA’s every 30 minutes during exercise. However, another similar study by Watson et al., (2004) where male participants cycled to volitional exhaustion at 50% VO2max in a warm environment consumed four aliquots of 250ml (3g) of a 12g/L-1 BCAA or placebo solution 30mins prior to exercise and 150ml every 15min during exercise. The participants had shown no effect in the delay of fatigue; placebo – 26.9min opposed to BCAA – 29.2min. Thus causing indefinite effects of BCAA supplementation and exercise capacity. 1.3 Physiological Affects The effects BCAA’s have on energy is approximately 3-18% of total energy. The amino acids; isoleucine, leucine and valine can also act as key components of the Krebs cycle to supply energy, making the three components of BCAA’s a valuable energy source (Cynober, 2013).The uses of BCAA’s have been shown to be effective predominantly with endurance- based activities and in terms of its physiological effects; one month’s oral supplementation of 9 essential amino acids, including BCAA’s, had increased fasting glucose and decreased creatine phosphokinase (a key enzyme which breaks down creatine and to resynthesise ATP and supply energy via the predominantly anaerobic ATP-PC energy system)activity in middle-long distance runners. Taken before and after workouts, marathoners as well as cyclists have shown positive effects during and before events where improvements in cognitive performance and reductions in time have been shown. Reductions in lactate, a key
  • 10. 10 component of exercise capacity and muscle mass loss has also shown to be reduced (Ohtani et al., 2001). BCAA’s enter the Krebs cycle directly as acetyl-CoA and not via the glycolytic pathway whereby pyruvate is converted to lactate by lactate dehydrogenase, a wasteful by-product that may inhibit exercise capacity (Harper et al., 1984). A study by Matsumoto et al., (2009) shows BCAA metabolism does not produce lactate and is considered to decrease lactate levels after supplementation. As lactate is a by-product of glycolysis during energy metabolism, ones lactate threshold and lactate concentration are considered as predictors of endurance exercise capacity (Yoshida et al., 1987). A study carried out by De Palo et al., (2001) reported lactate levels are suppressed within the blood during exercise following the chronic supplementation of BCAA’s. The study led by De Palo et al., (2001) had given triathletes 30 days of chronic BCAA supplementation (0.2g/kg-1) and 9.64g of BCAA oral- supplementation before exercise consisting of 60mins at 70% VO2max.Furthermore,a study carried out by Matsumoto et al., (2009) proved that 6 day supplementation of a BCAA drink increased lactate threshold by increasing workload levels at LT as well as VO2maxagainst a placebo. The study exercise test was an incremental loading exercise until exhaustion using a cycle ergometer; the test drink amounted to 500ml and was given 15min prior exercise. Participants took 1500ml/d during the 6 prior days to testing and it was concluded BCAA ingestion before exercise increases BCAA supply as an energy source to the muscle during endurance exercise. Thus it was theorised that the increase of acetyl-CoA to the Krebs cycle, via the BCAA catabolic pathway, inactivates the glycolytic pathway and consequently suppresses lactate production during the test. This suggests that a reduction in carbohydrate breakdown and lactate production increases endurance exercise capacity by inactivating the glycolytic energy system.
  • 11. 11 1.4 Consumption and Dosage On average, BCAA’s make up about 15% of total amino acid content within food protein (Gleeson, 2005). A Tour De France cyclist averages 25 Mj/d over a 2-3week period (Saris et al., 1989). Although protein in a tour de Frances’ diet may be relatively less, much energy is consumed in the form of carbohydrates, the protein is about 12% of total body energy for a tour de France cyclist. The elite cyclist consumes about 3000 kJ as protein thus about 19g of this protein are BCAA’s (Saris et al., 1989). Some studies have compared combined and separated glucose and BCAA supplementation (Madsen et al., 1996; Calders et al., 1999) and formulas are found to have both ergogenic aids within. The recommended dosage of BCAA’s is 3-20g a day, before and after workouts. A study carried out on 23 rugby players, who carried out intensive training, identifying the effects of a mixture of 9 essential amino acids, including leucine, isoleucine, valine and carbohydrates, showed significant improvements in vigour and earlier recovery from fatigue after 90 days of supplementation. They were given 3.6g twice, daily, for 90 days. This amount has been credited by most studies indicating 7-12g during endurance events, mixed into carbohydrate solution (Bloomstrand & Newsholme, 1992; Davis et al., 1999; Mero, 1999; Koba et al., 2007). A study by Tipton et al., (2004) showed whey protein ingestion resulted in a higher amount of blood BCAA concentrations, suggesting an increase in absorption when taken with whey protein or carbohydrates. Thus this study specifically looks at post lactate and glucose concentrations after BCAA supplementation as an indicator of endurance exercise capacity. However, dosage amount may critically vary amongst individuals which studies did not account for.
  • 12. 12 1.5 Lactate and Exercise Capacity The blood lactate curve and lactate thresholds have recently become an important factor in the assessment of exercise capacity (Faude et al., 2009). Lactate threshold, inversely related to blood lactate concentrations (bLa), has been always used as an index of endurance exercise capacity. In exercise intolerant mice with disrupted branched chain amino acid metabolism, increased rates of lactate release from skeletal muscle during exercise were described (She et al., 2010). It is a trend with many studies that graded incremental tests eliciting a rise in blood lactate concentration have been used to determine lactate thresholds or curves which indicate exercise capacity (Faude et al., 2009). During the first half of the 20th century VO2 max levels were the most common means of evaluating endurance capacity (Faude et al., 2009) and was developed by Hill et al., (1923). However, in the 1960’s the method by which lactate concentrations were measured was by capillary blood samples, which led to an increased popularity of bLa to assess endurance capacity (Hollmann, 2001) and has now become the most influential factor in the diagnosis of endurance performance/capacity in sport (Jones, 2006).Therefore this study uses bLa as a means of assessing exercise capacity and the improvements it may have due to BCAA supplementation, as it is generally accepted the lower the bLa concentration at a given workload the better ones endurance (exercise) capacity (Yoshida et al., 1990; Bosquet et al., 2002). 1.6Glucose and Exercise Capacity It has been outlined by Peronnet & Thibault, (1989) that the physiological basis of endurance capacity, determined by aerobic endurance, is not clearly grasped. It is, however, a combination of several factors one of which includes the capacity to spare carbohydrates in the form of glucose. The capacity to save carbohydrate as a reserve fuel by using more fatty acids as energy substrates increases ones (endurance) exercise capacity (Foster et al.,
  • 13. 13 1978).Thus this study uses the means of blood glucose levels (bGl) as a diagnosis of endurance-based exercise capacity. However, overall bLa are known to be influenced by depleted glycogen stores (proceeding exhaustive/endurance exercise) (Reilly & Woodbridge, 1999). For example, low bLa at the same work rates have been shown in glycogen-depleted subjects compared with a subject in normal condition. This may lead to a lower bLa and should not be interpreted as an increased exercise capacity (Maassen & Busse, 1989). Although studies suggest BCAA supplementation positively increases endurance exercise capacity, other studies have shown no effect as mentioned above by Watson et al., (2004) and as a result, disagreements remain with the effectiveness of BCAA ingestion on endurance exercise (Matsumoto et al., 2009). However, these studies used different timing and dosages thus affecting the comparison in relation to the effects of acute BCAA supplementation. Many studies have identified BCAA’s effects on central fatigue mechanisms opposed to peripheral fatigue mechanisms such as metabolism or fatigue. As no (Meta) analysis has been carried out to determine whether BCAA supplementation has an ergogenic effect on exercise capacity. This study looks at the effects BCAA supplementation has on metabolic (bGl) and fatigue (bLa) mechanisms in relation to exercise capacity.
  • 14. 14 2. Theoretical Framework 2.1 Research Questions Does BCAA supplementation have an effect on exercise capacity? 2.2 Research Aims To determine whether BCAA supplementation has an ergogenic effect on endurance-based exercise capacity. 2.3 Objectives To quantitatively review the research of BCAA supplementation and its effects on exercise capacity. H1- BCAA supplementation will enhance exercise capacity. H0 - BCAA supplementation will have no ergogenic effect on exercise capacity. 2.4 Operational Definitions Endurance exercise defined as any exercise mode in excess of 60 minutes work Work defined as an increase in HR due to physical activity Exercise capacity is defined as work until exhaustion measured by lactate or glucose
  • 15. 15 3. Methods 3.1 Literature Review and Study Selection The literature used within this meta-analysis was searched using the databases of GoogleScholar, SPORTDiscuss and PUBMED in July 2014. A variety of key search words and terms were used, these included but were not limited to: ‘BCAA’, ‘Branched-Chain- Amino-Acids’, ‘supplementation’, ‘exercise’, ‘capacity’, ‘endurance’, ‘lactate’, ‘glucose’. Once all potential studies had been identified, studies were included/excluded for analysis in accordance with the inclusion/exclusion criteria outlined below. Characteristics included relevant exercise modes such as endurance exercise and lactate/glucose measures. Homogeneity of participants was considered but inclusion criteria remained as outlined. 3.2 Inclusion/Exclusion Criteria This meta-analysis was limited to journal articles using human participants in a placebo or controlled double-blinded condition. These articles were published in English peer-reviewed journals which had a BCAA supplementation group, possibly including a carbohydrate solution and a placebo group (PLA). A summary of the processes used to select relevant articles to be used in the current meta-analysis are outlined in Fig. 1. Studies that were not included in the bLa analysis had no relevant info (Matsumoto et al., 2009; Greer et al., 2011; Pasiakos et al., 2011). Other studies that were not included had an inappropriate testing strategy (MacLean et al., 1996, where an anaerobic-based dynamic knee extensor exercise was used as a dependant variable), those with inappropriate experimental groups such as using rats or mice (Shimomura et al., 2000), or no specific numerical data in a table to use for analysis (Madsen et al., 1996; Coombes et al., 2000; Shimomura et al., 2004; Koba et al., 2007), or those that only seem to be available as abstracts (Varnier et al., 1994; Gualano et
  • 16. 16 al., 2011) and one that was only available in a Chinese journal hence was not accessible (Jianhong & Zhihong, 2005). 3.3 Data Analysis Pre- and Post-supplementation mean and standard-deviations (SD) were obtained from the original data from the studies used to establish effect size (ES).The effectiveness of the supplement was quantified by establishing the effects size(Δ) which was calculated by the difference between the post and pre-supplementation measures divided by the SD of the control group (Gene Glass’s approach). The control’s group standard deviation is used as it is not affected by the treatment (Glass et al., 1981).A forest plot as well as 95% confidence intervals (p≤0.05) displaying the findings of BCAA supplementation to be effective or not (on exercise capacity) was created by inputting raw data into Comprehensive Meta Analysis V2 software. Where shown in Fig. 1, n=number of study’s. Two out of six studies did not provide data indicating clear numerical lactate measurements, these data were instead given in a figure which was difficult to use for accurate lactate measures to use in this analysis, hence were excluded. Where information was not available, ‘no relevant data’ was stated, see Table 1.
  • 17. 17 Fig. 1. Flow chart showing the systematic review of studies and selection strategy Studiesthatwere excludeddue to irrelevantsample,predictors and outcomes(n=16) Studiesusedforfurtherevaluation (n=10) Total studies excluded(n=5) No relevantavailabledatafor lactate (n=2) Studiesincludedincurrent meta-analysis(n=5) Potential relevantstudies identified(n=26)
  • 18. 18 4. Results Table 1.Effects of BCAA supplementation on glucose (G) and lactate (L) levels on endurance-based exercise capacity. Study Sample Exercise Protocol Supplementation Protocol Total Dosage of BCAA (g) Effect Size Cheuvront et al., (2004) Watson et al., (2004) Matsumoto et al., (2009) 7 Healthy Males 8 Males 21 Trained Males 60min cycling @ 50% VO2 max. Then, 30min time-trial Cycling to volitional exhaustion @ 50% VO2 max. Incremental loading exercise test with cycle ergometer until exhaustion 60g/l glucose, 10g/l BCAA. Total of 1.4L; 200ml at 15min intervals 4 250ml aliquots of a 12g/l BCAA solution every 30min prior exercise for a total of 120min 150ml every 15min throughout exercise 6ml BCAA,60ml Carbohydrate solution; 1500ml/d for 6d 500ml 15min prior exercise test 24.4 54 2 (3.23-0.9)/1.27 ES = 1.83 (L) (6.15-5.02)/1.1 ES = 1 (G) (1.53- 0.59)/0.23 ES = 4.1 (L) (4.1-4)/1 ES = 0.1 (G) (114-101)/15 ES = 0.86(G) No Relevant info for Lactate Hsu et al., (2011) 14 Healthy Males 5min warm-up @ 55% VO2 max on a treadmill Then, ran @ 75% VO2 max for 30min. Thereafter,intensity increased by 1% until exhaustion 200ml of BCAA drink; valine (0.5g), leucine (1.0g), isoleucine (0.5g) and carbohydrate (12.1g) consumed post-exercise 2 (3.6-1)/0.4 ES = 6.5 (L) (80.3-78.1)/1.5 ES = 1.47 (G)
  • 19. 19 Study Sample Exercise Protocol Supplementation Protocol Total Dosage of BCAA (g) Effect Size Greer et al., (2011) Pasiakos et al., (2011) 9 Untrained Males 8 Adults 3 90min Cycling bouts @ 55% VO2 max Cycle ergometer @ 60% VO2 max BCAA drink; valine (7.3g), leucine (12.2g), isoleucine (4.8g). Administered 5min prior exercise and at 60min during exercise bout 125ml of 10g BCAA drink (leucine enriched) throughout exercise 24.3 10 (102.36- 102.34)/18.84 ES = 0.001 (G) No relevant info for Lactate (4.9-4.7)/0.1 ES = 2 (G) No relevant info for Lactate
  • 20. 20 Study name Statistics for eachstudy Stddiffinmeans and95%CI Stddiff Standard Lower Upper inmeans error Variance limit limit Z-Value p-Value Cheuvrontetal., (2004) 0.576 0.545 0.298 -0.493 1.645 1.056 0.291 Watsonetal., (2004) 0.050 0.500 0.250 -0.930 1.030 0.100 0.920 Matsumotoetal., (2009) 1.036 0.329 0.108 0.392 1.680 3.152 0.002 Hsuetal., (2011) 0.802 0.393 0.154 0.032 1.572 2.042 0.041 Greeretal., (2011) 0.002 0.471 0.222 -0.922 0.926 0.005 0.996 Pasiakos etal., (2011) 0.316 0.503 0.253 -0.670 1.302 0.629 0.530 0.576 0.178 0.032 0.227 0.925 3.237 0.001 -1.00 -0.50 0.00 0.50 1.00 Treatment effect p<0.05 Fig 2. Statistical findings of BCAA supplementation on Glucosemeasures Evaluation copy Studyname Statisticsfor each study Std diffin meansand 95% CI Std diff Standard Lower Upper in means error Variance limit limit Z-Value p-Value Cheuvront et al., (2004) 1.105 0.574 0.329 -0.020 2.230 1.926 0.054 Watson et al., (2004) 0.980 0.529 0.280 -0.057 2.017 1.852 0.064 Hsu et al., (2011) 1.554 0.431 0.186 0.709 2.399 3.603 0.000 1.269 0.289 0.083 0.703 1.835 4.394 0.000 -1.00 -0.50 0.00 0.50 1.00 Treatment effect p<0.05 Fig 3. Statistical findings of BCAA supplementation on Lactate measures Evaluation copy
  • 21. 21 5. Discussion and Conclusions This meta-analysis shows the overall effect of BCAA supplementation on endurance-based exercise capacity has a significant effect where p≤0.05; p=0.001 for glucose measures and p=0.000 for lactate measures (see fig. 2. and fig.3.). The analysis indicates that a dosage of 19.45g (mean dosage across all studies) of BCAA supplementation will have a positive effect on lower bLa and higher bGl indicating an enhanced exercise capacity for endurance-based activities. The results of this study are in agreement with previous studies including (Hsu et al., 2011; Matsumoto et al., 2011). In accordance to conventional effect size values, (Cohen, 1962) glucose may consider a ‘medium’ effect size for BCAA supplementation to be applicable and lactate may be considered to have a ‘large’ effect size; (d=0.032, d=0.083 respectively). The results for this study may be surprising as only two of six studies show significant differences between treatment and control groups in relation to lactate (Hsu et al., 2011; Motsumoto et al., 2011). However, the glucose trials identify significant differences which may be the reason for the significance of this study. It is important that these measurements are used together to determining the effects of BCAA supplementation as Cynober, (2013) states the use of BCAA’s should be in combination with carbohydrates. A study by Hsu et al., (2011) showed a BCAA drink containing 24.2g of carbohydrates may have enhanced the insulin response during post-exercise period. This suggests when BCAA and carbohydrates are mixed into a solution; it enhances anabolic responses during recovery (Hsu et al., 2011). Hsu et al., (2011) also found BCAA drink did not stimulate an increase in bLa, which is consistent with this study. The reason metabolism (of glucose) may be significant is the fact that BCAA’s may act as a substrate for muscle metabolism (Hsu et al., 2011). Amino acids have recently become more prominent in sport beverages due to an apparent reduction on muscle damage when consumed before or after exercise (Greer et al., 2007).
  • 22. 22 However, a concern arises for adding BCAA’s to spots drinks as it may add a potential carbon drain on the Krebs cycle. In the process of transimination, the first stop of BCAA catabolism, leucine is accepted and forms glutamate. When pyruvate (which forms lactate) is not available to supple alinine aminotransferase reaction, it may lead to a depletion of the Krebs cycle flux to produce ATP, during the oxidation of leucine, an essential amino acid (Wagenmakers, 1999). In conclusion the metabolic (bGl) and fatigue (bLa) mechanisms used as diagnosing exercise capacity has shown to be effective in enhancing exercise capacity by BCAA supplementation. Therefore the null hypothesis can be rejected in accordance with the above outlined glucose and lactate significance levels. 5.1 Limitations The results from this study do go against some previous studies such as (Watson et al., 2004; Cheuvont et al., 2004) but these studies were done less recently and thus may be irrelevant when compared with those that have been done more recently. Nonetheless, the studies included in this meta-analysis may show not significant measures compared with a placebo due to a better understanding of how to perform trials or a possible ‘placebo effect’ (Laursen et al., 2003). This study uses incremental exercise to exhaustion and exercise lasting 60mins or more to elicit rise in bLa. However, this measure of endurance capacity is hindered by many factors that may influence overall lactate levels. The magnitude of endurance gains is difficult to predict by using bLa (Faude et al., 2009) and may lead to this study to lack internal validity. Nevertheless, studies have shown bLa to be a renowned measurement in the diagnoses of exercise capacity (Yoshida et al., 1990; Bosquet et al., 2002; Faude et al., 2009).
  • 23. 23 6. Future recommendations In terms of the consumption of BCAA’s, they are safe within recommended dosages and have shown to improve exercise capacity within this meta-analysis if an average of 19.45g is taken according to the protocols used in this study. The use of amino acid supplements is not prohibited by the World-Anti-Doping Agency (WADA) and thus makes it a legal substance (Williams, 2005). This study proves the increased effects of endurance-based exercise capacity due to BCAA supplementation and thus may be an effective ergogenic aid for marathoners, cyclists and those involved in endurance activities. Coaches may use such supplements to quickly supply energy to athletes for endurance exercise as mentioned, BCAA’s predominantly metabolise in the muscle and fat tissue (Zeigler & Filler, 1996). However, the majority of participants in this study were healthy male adults and not elite athletes. The results in this meta-analysis were also calculated using comparatively few studies (n=6) which may make this study lack ecological validity. Furthermore, this study does not consider time-trial based results for measurements and although stated, does not take dosage effects into calculation. Findings of BCAA supplementation remain equivocal in particular its physiological effects on exercise capacity. Bloomstrand, (2001) reported reductions on perceived exertion ratings (RPE) and consequent central fatigue after BCAA supplementation during prolonged exercise, suggesting improvements in physical performance but the named study used central fatigue mechanisms to identify exercise enhancements. This highlights the need for future studies investigating the effects of BCAA supplementation on exercise capacity, in relation to peripheral fatigue mechanisms, is necessary with dosage, time-trial (performance), different mode(s) of exercise (muscular strength/power output) and different types as well as homogeneity amongst subjects as factors taken into consideration.
  • 24. 24 7. Critical reflection To undertake this research key life skills and technical skills had to be applied in order to successfully and efficiently carry out work. In doing so, time management skills to stick to deadlines had to be adhered to as well as excellent communication skills, which included emails to the supervisor, were made throughout this research project. Discipline to carry out work out of university hours was implied to keep up-to-date. The organisation of work was parallel to a successful project; organising relevant and irrelevant research papers into folders for future reference and usage within this analysis was very important to stay focused and precise in this project. Key transferable skills were also learnt and utilised such as the use of meta-analysis software (Comprehensive Meta Analysis V2), Microsoft Word, Outlook and PowerPoint for a high level of presentation of data, sending and constantly being aware of emails and presenting work in a slide show using PowerPoint. To be able to carry out this research project alongside other commitments such as a job has increased my ability to juggle many works at a single time and not only plan, but effectively manage time according to deadlines.
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