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Eur J Appl Physiol (2006) 97: 664672

DOI 10.1007/s00421-005-0036-1

O R I GI N A L A R T IC L E

Melissa J. Crowe Jarrad N. Weatherson


Bruce F. Bowden

Effects of dietary leucine supplementation on exercise performance

Accepted: 6 July 2005 / Published online: 29 October 2005


 Springer-Verlag 2005

Abstract Branched chain amino acids (BCAA), particularly leucine, have been suggested to be ergogenic for
both endurance and strength/power performance. This
study investigated the eects of dietary leucine supplementation on the exercise performance of outrigger
canoeists. Thirteen (ten female, three male) competitive
outrigger canoeists [aged 31.6 (2.2) year, VO2max 47.1
(2.0) ml kg 1 min 1] underwent testing before and after
6-week supplementation with either capsulated L-leucine
(45 mg kg 1 d 1; n=6) or placebo (cornour; n=7).
Testing included anthropometry, 10 s upper body power
and work and a row to exhaustion at 7075% maximal
aerobic power where perceived exertion (RPE), heart
rate (HR) and plasma BCAA and tryptophan concentrations were assessed. Leucine supplementation resulted
in signicant increases in plasma leucine and total
BCAA concentrations. Upper body power and work
signicantly increased in both groups after supplementation but power was signicantly greater after leucine
supplementation compared to the placebo [6.7 (0.7) v.
6.0 (0.7) W kg 1]. Rowing time signicantly increased
[77.6 (6.3)88.3 (7.3) min] and average RPE signicantly
decreased [14.5 (1.5)12.9 (1.4)] with leucine supplementation while these variables were unchanged with the
placebo. Leucine supplementation had no eect on the
plasma tryptophan to BCAA ratio, HR or anthropometric variables. Six weeks dietary leucine supplemenParts of this work have previously been presented in abstract form:
Crowe MJ, Weatherson JN (2002) The eects of dietary L-leucine
supplementation on exercise performance. Sports Medicine and
Science at the Extremes. Australian Conference of Science and
Medicine in Sport. 1216 October, Melbourne, Australia
M. J. Crowe (&) J. N. Weatherson
Institute of Sport and Exercise Science, James Cook University,
Townsville, QLD 4811, Australia
E-mail: Melissa.Crowe@jcu.edu.au
Tel.: +61-7-47815610
Fax: +61-7-47816688
B. F. Bowden
School of Pharmacy and Molecular Sciences,
James Cook University, Townsville, QLD 4811, Australia

tation signicantly improved endurance performance


and upper body power in outrigger canoeists without
signicant change in the plasma ratio of tryptophan to
BCAA.
Keywords Branched-chain amino acids Central
fatigue Supplements Tryptophan

Introduction
Research has shown that branched chain amino acids
(BCAA), particularly leucine, undergo increased oxidation during prolonged endurance exercise (Henderson
et al. 1985) and promote skeletal muscle protein synthesis (Layman 2002). For these reasons, there has been
interest in the potential of BCAA supplementation to
enhance both endurance and strength/power exercise
performance (Blomstrand et al. 1991, 1997; Davis et al.
1999; Mittleman et al. 1998; Pitkanen et al. 2003).
Research interest in BCAA has focussed on the
potential of the BCAA to prevent central fatigue
(Blomstrand et al. 1991, 1997) by competing with freetryptophan (f-TRP) for uptake from the plasma into the
brain. Increased brain f-TRP concentrations lead to increased serotonin (5HT) synthesis which is associated
with lethargy, fatigue and decreased exercise performance (see Davis et al. 2000). BCAA may limit central
fatigue as they compete with f-TRP for uptake into the
brain via the same transport mechanism (Pardridge and
Oldendorf 1975). Therefore, the ratio of plasma f-TRP
and BCAA may be important in preventing increases in
brain 5-HT and fatigue during exercise. Human and
animal studies support the central fatigue hypothesis
(see Davis et al. 2000). However, evidence that BCAA
supplementation can reduce central fatigue and improve
performance is less convincing. BCAA supplementation
studies have reported either decreased perceived exertion
and increased endurance performance (Blomstrand
et al. 1991, 1997; Mittleman et al. 1998) or no ergogenic
eects (Davis et al. 1999; Verger et al. 1994). The

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inconsistencies in BCAA supplementation studies may


relate to the level of carbohydrate ingestion which reduces the competition between plasma-free fatty acids
and f-TRP for the protein carrier, albumin, therefore,
inhibiting the increase in plasma f-TRP with exercise.
Thus, the potential for BCAA to enhance performance
above that achieved with carbohydrate alone has been
questioned (Davis et al. 1999; Madsen et al. 1996).
The few research studies that have investigated the
eects of BCAA or leucine supplementation on body
composition, strength or power have reported no eect
on anaerobic capacity, leg power or strength (Mourier
et al. 1997; Pitkanen et al. 2003). Abdominal fat was
reported to decrease after BCAA supplementation (Pitkanen et al. 2003) but muscle size was unaected when
BCAA supplementation was combined with other amino acids (Godard et al. 2002). Therefore, further studies
into the eects of BCAA on strength, power and body
composition are warranted.
The purpose of this study was to investigate the effects of dietary leucine supplementation on perceived
exertion and exercise endurance, anthropometric characteristics and upper body power of competitive outrigger canoeists. Outrigger canoeists were recruited to
the study as they commonly compete in both endurance
and sprint events, thereby providing an appropriate
population for evaluation of both endurance and power.
The leucine was provided as a daily dietary supplement
for 6 weeks to evaluate the eects on power and the
anthropometric characteristics as well as endurance
performance. It was hypothesised that leucine would
improve anthropometric characteristics, endurance and
power performance and reduce perceived exertion in
comparison to a placebo. It was also hypothesised that
leucine supplementation would increase performance via
an increase in plasma BCAA concentrations and a decrease in the plasma ratio of f-TRP to BCAA.

Methods
Subjects
Female (n=10) and male (n=3; one male participant
withdrew after injury unrelated to the study) outrigger
canoeists gave their written informed consent to participate in this study. The participant characteristics are
presented in Table 1. All the participants all had a

Table 1 Mean (SE) participant characteristics


Parameter
Number
Age (year)
Weight (kg)
Height (cm)
VO2max (mL kg

Males

3
27.3 (3.9)
96.3 (6.2)
185.9 (1.4)
min 1) 55.3 (1.4)

Females

All participants

10
32.9 (3.0)
67.5 (3.3)
166.8 (1.8)
44.6 (2.0)

13
31.6 (2.2)
74.2 (4.5)
171.2 (2.7)
47.1 (2.0)

minimum of 6 month outrigging experience and were


actively competing in regional outrigging canoeing
regattas on a monthly basis in both sprint (5003,000 m,
315 min) and marathon (1215 km, 12 h) events. All
participants underwent medical prescreening prior to
participation in the study and ethical approval to conduct the study was granted by the James Cook University Human Ethics Committee. The participants did not
take any medication during the study.
Experimental protocol
A randomised, double-blind design was employed
whereby the participants underwent pre-supplementation testing, a 6-week dietary supplementation period
and post-supplementation testing. The participants were
familiarised with each testing protocol and all testing
occurred at the same time of day pre- and post-supplementation. The participants were matched for age,
gender, mass and VO2max and randomly allocated to a
leucine (Musashi, Notting Hill, UK) supplementation
group (n=6; 45 mg kg 1 d 1) or a placebo group (n=7;
45 mg kg 1 d 1 cornour). Both the leucine and placebo were provided in 1,000 mg gelatin capsules and
consumed on a daily basis. To assess any potential adverse eects of supplementation, the participants were
verbally questioned about side eects during the postsupplementation testing session. The participants were
encouraged to maintain their normal training regime
throughout the testing and supplementation periods
with the exception of avoiding intense physical activity
during the 24 h prior to each testing session. Adherence
to this request was veried by the use of activity diaries.
Analysis of the activity diaries also showed that there
was no signicant dierence in the training volume between the participants in the leucine and placebo groups
in the week before pre-supplementation testing and
during the 6 weeks of supplementation (P=0.998). The
average duration of training was 3.7 (0.3) and 3.8
(0.2) h week 1 of outrigging training (the majority at
high intensity), plus 4.5 (2.1) and 4.3 (1.6) h week 1 of
cross training (resistance training, cycling, jogging) for
the leucine and placebo groups, respectively. In addition, all participants recorded their daily food intake
during the week before pre-supplementation testing and
throughout the supplementation period. Dietary analysis of daily protein intake was completed using Food
Wise software (McGraw-Hill, New York). The food
intake records also veried that the participants did not
consume any other supplements for the duration of the
study.
Testing commenced in the week prior to the supplementation period and included a VO2max test, upper
body 10 s power test, anthropometric characteristics and
a row to exhaustion at 7075% maximal aerobic power
with blood testing. All tests were repeated post-supplementation with the exception of the VO2max test. To
minimise any fatigue eects of the VO2max test on the

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power test and row to exhaustion at pre-supplementation, the anthropometry followed by the VO2max test
were performed on one day with the upper body power
test and row to exhaustion performed on a separate day,
17 days later. As the VO2max test was not repeated at
post-supplementation, the anthropometry was performed followed by the upper body power test and row
to exhaustion on the same day. At least 30-min rest was
provided between the 10 s upper body power test and
commencement of the row to exhaustion at both the preand post-supplementation sessions. Details of these
assessments are provided below.
VO2max assessment
Maximal oxygen consumption was determined using an
incremental rowing protocol modied from that of
Yoshiga and Higuchi (2003) on a rowing ergometer
(ConceptII, Morrisville, VT, USA). Female participants
commenced rowing at 100 W and increased the workload by 15 W every 2 min until volitional exhaustion.
Male participants underwent the same protocol with the
exception of commencing on 150 W with 50 W increments. All participants received verbal encouragement
throughout the duration of the test. Heart rate was recorded telemetrically (Polar Electro OY, Kempele,
Finland) every 30 s. The participants breathed through a
two-way valve (Hans Rudolph Inc.,Kansas City, MO,
USA) with gas analysis via a metabolic cart (Quinton
Instrument Company, Seattle, WA, USA) which was
calibrated with a 3 l syringe (Hans Rudolph Inc.,) and
standard alpha gas mixture (BOC Gases, Sydney, Australia). Oxygen consumption, CO2 production, respiratory exchange ratio (RER) and minute ventilation were
continuously recorded and averaged every 15 s of the
test. Verication of VO2max was established by the
achievement of three of the following four criteria: a
decrease or plateau in VO2 with an increase in the
workload; an RER greater than 1.15; HRmax > 85%;
and failure to maintain the specied workload (American College of Sports Medicine 2000).
Anthropometric characteristics
Anthropometric characteristics were determined in line
with the standards endorsed by the International Society
for the Advancement of Kinanthropometry (Norton and
Olds 1996) and performed by the same investigator
(technical error of measurement <5%). Height was assessed using a wall-mounted stadiometer and body mass
assessed using digital weighing scales (50 g; Precision
Health Scale VC-300, A & D Company Ltd, Japan).
Percentage body fat was calculated using LifeSize software (Version 1.0 Human Kinetics, Champaign, IL,
USA) from nine skinfold (bicep, tricep, subscapular,
iliac crest, supraspinale, abdominal, mid axilla, thigh
and calf), and ve girth (arm relaxed and exed, waist,

gluteal and calf) measurements. Skinfold and girth


measurements were performed using Harpenden skinfold callipers (British Indicators Ltd., West Sussex, UK)
and a metal tape measure (KDS Measure, Kyoto Measuring Instruments, Kyoto, Japan), respectively. The
mean of two measurements for each parameter was used
in the data analysis.
Upper body power and work
Relative peak power (W kg 1) and total work (J kg 1)
were determined via a 10 s arm crank test performed in a
standing position on a modied, air-braked cycle
ergometer attached to a power monitor (Supermonitor,
Repco Cycle Co., Huntingdale, Australia). Each subject
was tested in the same standing position at pre- and
post-supplementation with individual positions ranging
from shoulders level to shoulders 35 cm above the peak
of the crank rotation. Following a 5 min warm up at 50
100 W on the ergometer, the participants performed a
10 s all-out eort from a stationary start. Verbal
encouragement and time elapsed were provided
throughout the test.
Row to exhaustion
A fasting (10 h) pre-exercise blood sample (5 ml) was
taken from the antecubital vein and drawn into a lithium
heparin vacutainer that was stored at 4C. To simulate
competition conditions, the participants were provided a
pre-exercise meal that consisted of a carbohydrate/electrolyte drink (Powerade) and fruit bars (Uncle Tobys
Real Fruit Bars) to deliver approximately 1 g kg 1
carbohydrate (Burke 2000). The last dose of leucine or
placebo was consumed with the drink after the blood
sampling. The need for a fasting blood sample therefore
resulted in a period of approximately 24 h between the
previous intake of supplement and blood sampling.
Twenty minutes after ingestion of the carbohydrate
meal, the participants commenced a 5-min warm up on
the rowing ergometer at 80120 W. Following the warm
up, the participants performed a row to exhaustion at
7075% maximal aerobic power as determined from the
VO2max test with HR and rating of perceived exertion
(RPE; Borg 1978) recorded every 5 min. To simulate
competition conditions during the row, the participants
were provided a diluted carbohydrate/electrolyte drink
(Powerade, 4% carbohydrate) ad libitum using a handsfree backpack system. The drink volume consumed
during the row was recorded. The participants had visual access to power output (W) and were provided
verbal encouragement throughout the row. Exhaustion
was determined upon failure to maintain 7075% maximal power output in excess of 10 s. Upon completion of
exercise, time to exhaustion was recorded and a postexercise blood sample obtained in the same manner as
the pre-exercise sample.

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Blood testing
Blood samples were analysed using high performance
liquid chromatography (HPLC) for plasma concentrations of leucine, isoleucine, valine and tryptophan.
Within 4 h of collection, the blood samples were centrifuged (3,000 rpm for 25 min at 4C) and the plasma
frozen at 20C for later analysis. Following thawing,
plasma preparation was carried out in accordance with
the methods of Gratseld-Hueesgen (1998) to precipitate
protein and extract lipophilic materials. To precipitate
proteinaceous material, 2 ml of methanol was added to
the samples with 4 ml of 1:1 diethyl ether and hexane to
extract lipophilic materials. The sample was then mixed
in a vortex for 60 s and centrifuged (3,000 rpm for
10 min at 4C). After centrifugation, the upper hexane/
diethyl ether phase was removed and discarded. A further 4 ml of the diethyl ether/hexane mixture with 1 ml
of methanol was added and the sample vortexed and
centrifuged (3,000 rpm for 15 min at 4C). The supernatant was removed and retained. The pellet was washed
twice more with 1 and 0.5 ml methanol and then discarded. The sample was divided into two 2 ml aliquots
that were snap-frozen using liquid nitrogen and placed
on a freeze drier overnight.
Amino acid derivatisation and HPLC analysis were
performed with modications to the methods of Carroll
et al. (1996). Following freeze-drying, the aliquots were
treated with 500 ll distilled water. A 200 ll aliquot was
then derivatised with 2.5 mg of 1-uoro-24-dinitophen5-yl-L-alanine amide (FDAA) in 250 ll of acetone and
1N sodium bicarbonate (30 ll) at 50C for 4 h. Samples
were acidied with 125 ll hydrochloric acid and ltered
through 0.22 ll lters (Millipore Corporation, Bedford,
MA, USA) to remove all the remaining proteins and
transferred into 2 ml glass ampoules. High performance
liquid chromatography analysis was performed on a
Reverse Phase Luna phenyl-hexyl column (4.6250 mm;
Phenomonex, Auckland, New Zealand) with an autosampler injector (ICI Instruments, Hubbartson, MA,
USA). The best conditions for providing separation
were found to be a mobile phase of 72:28% triethylammonium phosphate (pH 3.0):acetonitrile, at a ow
rate of 2.0 ml min 1 and UV detection at 340 nm.
Each peak in the chromatographic trace was identied
by comparing the retention time with that of standards of
L-leucine, L-isoleucine, L-valine and L-tryptophan (British
Drug Houses Ltd., Poole, UK). The standards were
prepared by serial dilution and derivatised, acidied and
run on the column in the same manner as the plasma
samples. The concentrations of each amino acid in the
plasma were determined by comparing the integral of the
peak with that of the standard.

data was analysed using an independent samples t test


(Howell 1992). The dietary protein intake, exercise volume, anthropometry, upper body peak power and total
work, exercise time to exhaustion and RPE at exhaustion
data were analysed using a 22 (grouptime) factorial
ANOVA with repeated measures for time (Howell 1992).
Heart rate and RPE during the row to exhaustion and the
plasma parameters (leucine, isoleucine, valine, tryptophan, BCAA:tryptophan ratio) were analysed using a
3-way factorial ANOVA (timegroupexercise) with repeated measures for time and exercise. The Greenhouse
Geiser correction was used where assumptions of
sphericity were not met (Howell 1992). Post hoc analysis
was performed using the Tukey HSD test. Due to an
unequal sample size, the harmonic mean of the samples
sizes was used in place of n in the Tukey test (Howell
1992). Statistical power is reported for each of the variables that changed signicantly and data are presented as
mean (SE). Alpha was set at 0.05.

Results
VO2max
There was no signicant dierence in the VO2max of the
two groups prior to supplementation (P=0.681) with
the leucine and placebo groups averaging 46.1 (3.3) and
47.9 (2.7) mL kg 1 min 1, respectively.
Dietary analysis
The dietary protein intake of the leucine and placebo
groups did not dier signicantly during the supplementation period [leucine group 0.85 (0.06) g kg 1 d 1;
placebo group 0.85 (0.05) g kg 1 d 1; P=0.775]. Furthermore, dietary intake of protein did not change from
the week preceding pre-supplementation testing to the
supplementation period for either group (P=0.570).
Anthropometric characteristics
There was no signicant dierence between the leucine
and placebo groups in body mass at pre-supplementation [leucine 73.0 (5.1) kg, placebo 75.2 (7.5) kg] or postsupplementation [leucine 73.1 (5.0) kg, placebo 75.1
(7.5) kg; P>0.05]. Similarly, percentage body fat was
not signicantly dierent between the leucine and placebo groups at pre-supplementation [leucine 25.1
(3.9)%, placebo 26.4 (1.6)%] or post-supplementation
[leucine 25.7 (3.6)%, placebo 26.6 (1.6)%; P>0.05].

Statistical analysis

Upper body power and work

All data were analysed using SPSS for Windows software


(Version 11; SPSS Inc., Chicago, II, USA). The VO2max

Relative peak power signicantly increased from pre- to


post-supplementation for both the leucine and placebo

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groups (P<0.001; power=1.00), however, the postsupplementation peak power achieved by the leucine
group was signicantly higher than that achieved by the
placebo group (P=0.045; power=0.54; Fig. 1a). Total
work achieved during the 10-s-test also increased signicantly for both groups from pre- to post-supplementation (P=0.001; power=0.99). However, there was
no signicant dierence between the leucine and placebo
groups at post-supplementation (P=0.277; Fig. 1b).
Row to exhaustion
A main eect for time showed that regardless of supplementation group, there was an overall improvement
in the rowing time to exhaustion from 72.2 (4.4) min at
pre-supplementation to 76.3 (5.8) min at post-supplementation (P=0.036; power=0.59). The interaction effect (P=0.008; power=0.84) showed a signicant
increase in time to exhaustion for the leucine group
whereas the placebo group showed no improvement
(Fig. 2).
Exercise time to exhaustion varied between 55 and
110 min. Therefore, to analyse a complete data set for
all participants, only the rst 55 min of RPE and HR
data were analysed. There was a signicant main eect
for time on RPE with a signicant increase in RPE
every 10 min throughout the row to exhaustion test
(P<0.001; power=1.00). The average RPE was not
signicantly dierent between the leucine and placebo
groups at pre-supplementation [14.5 (1.5) and 14.6
(1.2), respectively]. However, the average RPE for the
leucine group was signicantly lower than that of the
placebo group during the post-supplementation row to

Fig. 1 Upper body a peak power and b total work for the leucine
and placebo groups at pre- and post-supplementation. Data are
mean (SE). *P<0.05 signicantly dierent from leucine at presupplementation and placebo at pre- and post-supplementation

Fig. 2 Row time to exhaustion for the leucine and placebo groups
at pre- and post-supplementation. Data are mean (SE). *P<0.05
signicantly dierent from leucine at pre-supplementation and
placebo at pre- and post-supplementation

exhaustion [12.9 (1.4) and 15.0 (1.4), respectively;


P=0.031; power=0.62]. A signicant 3-way interaction eect showed that the RPE for the leucine group
at 25 and 50 min of the row was signicantly lower at
post-supplementation compared to the same times
during pre-supplementation testing (P=0.001; power=0.98; Fig. 3). This interaction eect also showed
that the RPE of the leucine group was signicantly
lower than that of the placebo group at all times between 30 and 55 min of the row at post-supplementation (Fig. 3). The average RPE at exhaustion was
not signicantly dierent between the leucine and
placebo groups at pre-supplementation [leucine, 19.7
(0.3); placebo 19.7 (0.2)] or post-supplementation
testing [leucine, 19.5 (0.4); placebo, 19.6 (0.4);
P=0.968].
The participants maintained the prescribed exercise
intensity during the row to exhaustion with no signicant dierence in the average exercise HR between the
two groups during either pre-supplementation [leucine,

Fig. 3 Rating of perceived exertion (RPE) for the leucine and


placebo groups at pre- and post-supplementation. Data are mean
(SE). Plac placebo; Leu leucine; Pre pre-supplementation; Post
post-supplementation. *P<0.05 leucine signicantly dierent from
placebo at post-supplementation;# P<0.05 signicantly dierent
from pre-supplementation for the leucine group

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Table 2 Mean (SE) plasma amino acid concentrations for all participants (n=13) prior to and following the row to exhaustion
Amino acid (lmol)

Pre-exercise

Post-exercise

Leucine
Isoleucine
Valine
f-tryptophan
Total BCAA
f-TRP:BCAA

145.1 (4.0)
76.8 (2.3)
262.0 (4.6)
13.4 (0.8)
483.9 (10.4)
0.0280 (0.0020)

143.2 (4.0)*
75.0 (2.2)*
250.0 (4.5)*
14.0 (0.7)*
478.2 (10.3)*
0.0294 (0.0017)*

*P<0.05 signicantly dierent from pre-exercise

158.2 (9.3) beats min 1; placebo, 161.8 (7.0) beats


min 1] or post-supplementation testing [leucine, 156.3
(9.8) beats min 1; placebo 161.5 (6.6) beats min 1; P=
0.508].
The volume of carbohydrate/electrolyte solution ingested during the row to exhaustion was not signicantly dierent between the leucine and placebo groups
at pre- [leucine, 261 (40) ml; placebo 267 (37) ml] or
post-supplementation testing [leucine, 264 (41) ml;
placebo, 261 (38) ml; P=0.800].
Blood parameters
There were no signicant dierence between the leucine
and placebo groups in any of the plasma amino acid
concentrations prior to supplementation (P>0.05).
There was a signicant main eect for exercise on the
plasma concentrations of each BCAA, f-tryptophan,
total BCAA concentrations and the ratio of f-tryptophan to BCAA (Table 2). The exercise resulted in a
signicant decrease in plasma leucine (P<0.001; power=1.00), isoleucine (P<0.001; power=1.00), valine
(P<0.001; power=1.00) and total BCAA (P<0.001;
power=1.00; Table 2). In contrast, the exercise resulted
in a signicant increase in plasma f-tryptophan
(P=0.037; power=0.58) and the ratio of f-tryptophan
to BCAA (P=0.011; power=0.79; Table 2).
A main eect for time showed that regardless of
supplementation group, there was an overall increase in
plasma leucine concentrations following supplementation [pre-supplementation, 139.4 (4.5) lmol; post- supplementation 148.9 (3.6) lmol; P=0.0002; power=
1.00]. A signicant interaction eect showed a signicant
increase in plasma leucine concentrations for the leucine
group from pre- to post-supplementation but no significant change for the placebo group (P<0.001; power=0.99; Fig. 4).
Plasma isoleucine concentrations did not change
signicantly from pre- to post-supplementation in either
the leucine or placebo groups (P=0.369; Table 3). A
signicant 3-way interaction eect indicated a signicant
decrease in plasma isoleucine concentrations from
pre- to post-exercise for the leucine group at pre-supplementation and the placebo group at post-supplementation (P=0.042; power=0.55; Table 3). However,
there were no signicant dierence in the plasma

Fig. 4 Plasma leucine concentrations for the leucine and placebo


groups at pre- and post-supplementation prior to and following
exercise. Data are mean (SE). *P<0.05 signicantly dierent from
leucine at pre-supplementation and placebo at pre- and postsupplementation

isoleucine concentrations between the leucine and placebo groups pre- or post-exercise following the supplementation period (Table 3).
Plasma valine concentrations did not change signicantly from pre- to post-supplementation in either the
leucine or placebo groups (P=0.632; Table 3). Similarly, plasma f-tryptophan concentrations were unaffected by leucine supplementation with no signicant
dierences from pre- to post-supplementation in the
leucine or placebo groups (P=0.671; Table 3).
The total BCAA concentrations showed a signicant
main eect for time with a signicant increase from preto post-supplementation [476.5 (11.5) lmol v. 485.5
(9.2) lmol; P=0.014; power=0.75]. A condition by
time interaction showed that BCAA were signicantly
increased from pre- to post-supplementation in the leucine group [479.3 (16.9) v. 497.4 (13.5) lmol, respectively] and that the post-supplementation levels in the
leucine group were signicantly greater than those of the
placebo group at both pre- and post-supplementation
[473.7 (15.7) v. 473.6 (12.5) lmol, respectively;
P=0.014; power=0.76].
The ratio of f-tryptophan to BCAA was unaected
by leucine supplementation with no signicant dierence
between the leucine and placebo groups at pre- or postsupplementation before or after exercise (P=0.438;
Table 3).
Side eects
None of the participants reported any side eects as a
result of dietary supplementation.

Discussion
The results of this study supported the hypotheses that
leucine supplementation would increase plasma leucine

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Table 3 Mean (SE) plasma concentrations of isoleucine, valine, free-tryptophan (f-TRP) and the ratio of f-TRP to BCAA for the leucine
and placebo groups

Pre-supplementation
Leucine (Pre-exercise)
Leucine (Post-exercise)
Placebo (Pre-exercise)
Placebo (Post-exercise)
Post-supplementation
Leucine (Pre-exercise)
Leucine (Post-exercise)
Placebo (Pre-exercise)
Placebo (Post-exercise)

Isoleucine (lmol)

Valine (lmol)

f-TRP (lmol)

f-TRP:BCAA

76.7
74.5
77.4
76.1

(3.4)
(3.5)*
(3.1)
(3.2)

263.7
262.2
260.4
257.6

(6.7)
(6.8)
(7.4)
(7.0)

13.5
14.3
14.1
14.4

(1.5)
(1.3)
(1.5)
(0.9)

0.028
0.030
0.029
0.031

(0.029)
(0.030)
(0.040)
(0.032)

76.3
75.0
76.9
74.3

(3.3)
(3.2)
(3.1)
(2.9)*

263.0
261.3
260.7
258.9

(5.8)
(5.7)
(6.1)
(8.0)

13.0
13.2
13.4
13.9

(0.9)
(0.7)
(0.9)
(0.6)

0.026
0.027
0.029
0.030

(0.022)
(0.018)
(0.025)
(0.017)

*P<0.05 signicantly dierent from corresponding pre-exercise concentration

and total BCAA concentrations, and improve upper


body power output, RPE and exercise time to exhaustion in outrigger canoeists. However, the data suggest
that the increased performance following leucine supplementation was not likely to be mediated by a reduction in central fatigue as there was no signicant
reduction in the plasma ratio of f-TRP to BCAA.
The results of the current study are in agreement with
past studies (Rennie et al. 1981; Watson et al. 2004) with
signicant decreases in leucine, isoleucine and valine
concentrations and a signicant increase in f-TRP following exercise, regardless of supplementation with either placebo or leucine. A small number of past studies
(Blomstrand et al. 1991; Mittleman et al. 1998) have
shown that supplementation with BCAA can limit the
decrease in plasma BCAA concentrations during exercise and thus prevent the increase in the ratio of f-TRP
to BCAA. Leucine supplementation in the current study
resulted in a signicant elevation in the plasma concentrations of leucine and total BCAA. Furthermore,
RPE during the row to exhaustion was also signicantly
decreased in the leucine but not the placebo group following supplementation. Both these results indicate a
reduction in central fatigue. A previous investigation
into the eects of BCAA on perceived exertion during
exercise suggested that BCAA had a central eect which
resulted in reduced RPE and mental fatigue (Blomstrand
et al. 1997). In further support of a central mechanism,
Blomstrand et al. (1997) reported that during exercise
the plasma ratio of f-TRP to BCAA was maintained at
pre-exercise concentrations or decreased. However, in
contrast to the Blomstrand et al. (1997) study, leucine
supplementation in the current study did not signicantly alter the exercise-induced increase in plasma fTRP or the ratio of f-TRP to BCAA. Therefore, the
plasma ratio of f-TRP to BCAA may not have played a
role in mediating the ergogenic eects of leucine in the
current study.
Previous studies investigating leucine and BCAA
supplementation with endurance performance have
shown equivocal results (Blomstrand et al. 1991, 1997;
Mittleman et al. 1998; Verger et al. 1994; Watson
et al. 2004). Methodological dierences between these

studiesparticularly with reference to the exercise protocol, BCAA dose and carbohydrate availabilitymake
comparisons dicult. Past BCAA supplementation
studies have reported an increase in marathon performance in slower but not quicker runners (Blomstrand
et al. 1991) and an increase in exercise time to exhaustion in the heat (Mittleman et al. 1998). The latter study
reported a decrease in concentrations of f-TRP and the
ratio of f-TRP to BCAA in response to BCAA supplementation (Mittleman et al. 1998). In contrast, glycogen-depleted subjects who exercised to exhaustion in the
heat did not benet from BCAA supplementation
(Watson et al. 2004). Plasma ammonia concentrations
were also elevated following exercise with BCAA in this
study (Watson et al. 2004). The dose of BCAA has been
implicated in the outcome of BCAA supplementation
studies with the suggestion that higher doses increase
plasma ammonia concentrations and therefore result in
fatigue (Blomstrand 2001). Consistent with this suggestion, the current study utilised a dose of approximately
3.3 g d 1 and other studies reporting positive eects of
BCAA supplementation have utilised doses of approximately 68 g (Blomstrand et al. 1997) and 9 and 16 g
(Mittleman et al. 1998). In contrast, those studies
reporting no eect of BCAA supplementation on performance utilised approximately 1730 g BCAA (Madsen et al. 1996; Varnier et al. 1994; Watson et al. 2004).
However, one of these studies (Varnier et al. 1994) did
not detect a dierence in plasma ammonia concentrations between the BCAA and placebo treatments. A
dose-response study of BCAA supplementation and
subsequent alterations in plasma BCAA, the ratio of
f-TRP to BCAA and ammonia may assist in clarifying
the exercise response to BCAA supplementation.
Supplementation of BCAA with carbohydrate has
previously been reported to provide no additional performance benets compared to carbohydrate supplementation alone (Davis et al. 1999; Madsen et al. 1996).
The current study provided a pre-exercise carbohydraterich meal in order to simulate competition conditions. It
was therefore interesting that after 6-week leucine supplementation, endurance performance was improved to
a greater extent than after the placebo where the same

671

dose of pre-exercise carbohydrate was ingested. The


improvement in endurance performance with leucine
supplementation despite carbohydrate would indicate
that these eects were mediated by a mechanism other
than a reduction in central fatigue.
In contrast to the current study, where leucine was
administered chronically for a 6-week period, the
majority of past endurance exercise studies have
administered single or multiple doses of the BCAA
supplement on the day of exercise testing (Blomstrand
et al. 1991, 1997; Madsen et al. 1996; Mittleman et al.
1998; Watson et al. 2004). The last dose of leucine/placebo in the current study was administered prior to
exercise testing with ingestion of the carbohydrate meal.
The signicant increase in plasma leucine concentrations
in the current study was of much lower magnitude [14.0
(3.2)%] than other studies (100%; Mittleman et al.
1998; Watson et al. 2004). It is therefore not surprising
that the ratio of f-TRP to BCAA was not signicantly
altered. Supplementation for a chronic 6-week period
would have increased the daily dietary intake of protein
for the participants in the leucine group compared to
those in the placebo group. Both groups averaged a
daily protein intake of 0.85 g kg 1 from dietary sources
which is relatively low for endurance athletes who are
usually advised to consume approximately 1.2 g kg 1
protein per day (Tarnopolsky 2000). However, the dose
of leucine consumed would only have increased the total
daily protein intake by approximately 5%. Therefore,
the dierence in daily protein intake between the leucine
and placebo groups over the 6-week supplementation
period may have had a small eect but is unlikely to
fully explain the ergogenic eects of leucine in this study.
In contrast with the majority of past studies investigating strength, power or anaerobic performance
(Godard et al. 2002; Mero et al. 1997; Mourier et al.
1997; Pitkanen et al. 2003), the results of the current
study showed a signicant increase in upper body
power after supplementation with leucine. Wingate test
performance and knee extensor strength in wrestlers
(Mourier et al. 1997), muscle size and strength in older
men (Godard et al. 2002) and countermovement jump
performance in power athletes (Mero et al. 1997) were
unaected by varying periods of dietary leucine or
BCAA supplementation when compared to placebo
conditions. An acute dose of BCAA prior to the
exercise also had no eect on maximal anaerobic running or countermovement jump performance (Pitkanen
et al. 2003). Leucine supplementation in the current
study had mixed eects on power performance with a
signicant increase in peak power but no eect on the
total work achieved in the 10 s arm crank test. Direct
comparisons between studies into the eects of BCAA
on strength, power and anaerobic performance are
dicult because of the variation in supplement mixture,
population utilised and duration of supplementation.
The Mero et al. (1997) study utilised a similar dose and
time period of leucine supplementation to that of
the current study but reported no improvement in

countermovement jump performance in power athletes.


Although this study tested only male athletes (Mero
et al. 1997), the majority of participants in the study
were female. Males have previously been reported to
oxidise leucine at a greater rate during exercise compared to females (Lamont et al. 2001). Nonoxidative
disposal of leucine has also been reported to be signicantly lower in males compared to females leading
to a diminished rate of protein synthesis during exercise
in men (Lamont et al. 2001). Comparison of gender
dierences in response to leucine was not possible in
the current study due to the low number of male participants. However, it could be speculated that over a
6-week period, greater benets from training occurred
in the current study as the majority of participants
were female compared to past studies which only utilised males. However, the total number of participants
in the current study was low and these results should
be replicated in a larger sample with particular
emphasis on gender dierences and long-term training.
The mechanism underlying the improvement in
endurance and upper body power with leucine supplementation is unclear. The signicant increase in plasma
leucine and total BCAA concentrations and signicant
decrease in exercise RPE after leucine supplementation
indicate a decrease in central fatigue. However, the lack
of change in the ratio of f-TRP to BCAA with leucine
supplementation does not support this hypothesis.
Alternative leucine mechanisms may involve direct effects of leucine on skeletal muscle. Past studies have
reported that leucine signicantly increased skeletal
muscle protein synthesis (see Layman 2002) and that
BCAA supplementation reduced serum indicators of
muscle damage following prolonged endurance exercise
(Coombes and McNaughton 2000). It is therefore possible that 6-week dietary leucine supplementation in the
current study reduced training-induced muscle damage
and enhanced recovery resulting in improved benets
from training and increased performance. Alternatively,
BCAA supplementation has been hypothesised to alter
plasma concentrations of anabolic hormones (Coombes
and McNaughton 2000). However, increases in anabolic
hormone concentrations are unlikely to have mediated
the ergogenic eects of leucine in the current study as the
anthropometric characteristics did not show any anabolic eects. Although the authors acknowledge the
limitations of using anthropometry to detect changes in
body composition, the body mass and percentage body
fat data indicated no signicant change over the 6-week
period in either group. Furthermore, testosterone and
human growth hormone (hGH) responses to BCAA or
leucine supplementation are not well characterised (Carli
et al. 1992; Mero et al. 1997). Therefore, the ergogenic
eects of leucine were more likely related to a reduction
in skeletal muscle damage with training and/or an increase in skeletal muscle protein synthesis. Future research should further explore the mechanisms by which
leucine or BCAA can prevent muscle damage and increase muscle synthesis.

672

In conclusion, upper body power, time to exhaustion


and perceived exertion were signicantly improved after
6-week dietary leucine supplementation compared to a
placebo. Although leucine supplementation signicantly
elevated plasma leucine and total BCAA concentrations,
there was no signicant dierence in the elevation in
plasma f-TRP and the ratio of f-TRP to BCAA with
exercise. Therefore, dietary leucine supplementation was
eective at improving performance but this eect was
unlikely to have been mediated by a reduction in central
fatigue. Future studies should further examine the
capacity of leucine to improve skeletal muscle synthesis
and reduce muscle damage during training and investigate gender dierences in response to leucine supplementation during long-term training.

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