J Exerc Nutr Biochem 2013;17(4):169-180

ISSN : 2233-6834 (Print) ISSN : 2233-6842 (Online)

http://dx.doi.org/10.5717/jenb.2013.17.4.169

ORIGINAL PAPER
*

Effect of BCAA intake during endurance exercises on fatigue substances, muscle

damage substances, and energy metabolism substances
Dong-Hee Kim1, Seok-Hwan Kim1*, Woo-Seok Jeong1 and Ha-Yan Lee2
1
Department of Physical Education, Chonnam National University, Gwangju, Korea
2
Department of Sports Leisure, Nambu university, Gwangju, Korea

(Received: 2013/10/30, Revised: 2013/11/19, Published online: 2013/11/28)

ABSTRACT
The increase rate of utilization of branched-chain amino acids (BCAA) by muscle is reduced to its plasma concentration during
prolonged exercise leading to glycogen. BCAA supplementation would reduce the serum activities of intramuscular enzymes associated
with muscle damage. To examine the effects of BCAA administration on fatigue substances (serotonin, ammonia and lactate), muscle
damage substances (CK and LDH) and energy metabolism substances (FFA and glucose) after endurance exercise. Subjects (n =
26, college-aged males) were randomly divided into an experimental (n = 13, EXP) and a placebo (n = 13, CON) group. Subjects
both EXP and CON performed a bout of cycle training (70% VO2max intensity) to exhaustion. Subject in the EXP were administrated
BCAA (78ml/kg·w) prior to the bout of cycle exercise. Fatigue substances, muscle damage substances and energy metabolism substances
were measured before ingesting BCAAs and placebos, 10 min before exercise, 30 min into exercise, immediately after exercise,
and 30 min after exercise. Data were analyzed by two-way repeated measure ANCOVA, correlation and statistical significance was
set at p < 0.05. The following results were obtained from this study; 1. In the change of fatigue substances : Serotonin in the EXP
tended to decreased at the 10 min before exercise, 30 min into exercise, post exercise, and recovery 30 min. Serotonin in the CON
was significantly greater than the EXP at the10 min before exercise and recovery 30. Ammonia in the EXP was increased at the
10 min before exercise, 30 min into exercise, and post exercise, but significantly decreased at the recovery 30min (p < 0.05). Ammonia
in the CON was significantly lower than the EXP at the 10 min before exercise, 30 min into exercise, and post exercise (p < 0.05).
Lactate in the EXP was significantly increased at the 30 min into exercise and significantly decreased at the post exercise and
recovery 30 min. Lactate in the CON was significantly lower than the EXP at the post exercise (p < 0.05). 2. In the change of
muscle damage substances : CK in the EXP was decreased at the 10 min before exercise and increased at the 30 min into exercise
and then decreased at the post exercise and recovery 30 min. CK in the CON was greater than the EXP. LDH in the EXP was
decreased at the 10 min before exercise and increased at the 30 min into exercise and then decreased at the post exercise and
recovery 30 min. LDH in the CON was higher than the EXP. 3. In the change of energy metabolism substances :Glucose in the
EXP tended to decrease at the 10 min before exercise, 30 min into exercise, post exercise and recovery 30 min. Glucose in the
CON was significantly greater than the EXP at the recovery 30 min (p < .05). FFA in both EXP and CON was increased at the
post exercise and recovery 30 min. % increase for FFA in the EXP was greater than the CON at the post exercise and recovery
30 min. 4. The relationship of the fatigue substances, muscle damage substances and energy metabolism substances after endurance
exercise indicated strongly a positive relationship between LDH and ammonia and a negative relationship between LDH and FFA
in the EXP. Also, there were a strong negative relationship between glucose and FFA and a positive relationship between glucose
and serotonin in the EXP. There was a strong positive relationship between CK and LDH and a strong negative relationship between
FFA and glucose in the CON.
These results indicate that supplementary BCAA decreased serum concentrations of the intramuscular enzymes as CK and LDH
following exhaustive exercise. This observation suggests that BCAA supplementation may reduce the muscle damage associated with
endurance exercise.

Keywords: BCAA, fatigue substances, muscle damage substances, energy metabolism substances

* Corresponding author: Seok-Hwan Kim, Tel. 82-62-513-5636, Email. [email protected]

ⓒ2013 The Korean Society for Exercise Nutrition
170 BCAA intake during endurance exercise

INTRODUCTION exercise begins and glycogen stored in the liver is mobilized.

However, when BCAA is ingested, the BCAA moves to the
Recently, ordinary people as well as athletes consume more muscle and is oxidized to supply additional energy. As a
and diverse ergogenic aids to improve density of skeletal result, the amount of glucose that is decomposed in the liver
muscle and exercise performance [2]. As prolonged exercise and released into the blood is reduced and accordingly, the
depletes carbohydrate and mobilizes fat as an energy source, level of glucose in the blood is decreased. It was reported
fat acts as an important energy substrate for muscle function that the intake of BCAA helped to prevent physical
[3]. In order to improve exercise performance, carbohydrates performance from deteriorating, which is usually caused by
and amino acids which are the source of protein, especially muscle glycogen depletion in the later stages of endurance
BCAA (Branched Chain Amino Acid; BCAA) are critical. exercises [3].
Amino acids help enhance metabolism which promotes cell However, there is no enough research for the BCAA dosage
proliferation and improve exercise performance as well as depending on forms of exercise, as well as changes in fatigue
functional recovery during exercise [18,8]. Also, they help substances, muscle damage substances, and energy meta-
perform better by contributing as the energy source and bolism substances. Thus, this study was conducted not only
reducing the accumulation of 5-HT (serotonin), the central to analyze how the intake of BCAA during endurance exercise
fatigue substance. Blood CK (creatine kinase; CK) concen- affects fatigue substances, muscle damage substances, and
tration and blood LDH (lactate dehydrogenase; LDH) energy metabolism substances but also to identify interrelated
concentration are the indicators that reflect the degree of factors.
muscle damage and physical fitness from long-term physical
activity [19]. CK is the main enzyme that controls the
ATP-PC system and LDH is the main enzyme that maintains RESEARCH METHOD
the balance of sugar catabolism and anabolism. Coombes and
McNaughton[4]. reported that CK and LDH concentrations
Research subjects
had decreased after they exercised on a bicycle ergometer
with an intensity of 70 percent of their maximal oxygen In this study, 30 male subjects were primarily selected from
uptake for a long period of time after following a common student volunteers attending J University. The 30 subjects then
diet and taking 64mg of BCAA in proportion to body weight. had to undergo a physical examination to determine whether
Greer[7]. reported that those who exercise on a bicycle they had metabolic anomalies, cardiovascular anomalies, or
ergometer with an intensity of 50 percent of their maximal any disorder associated with muscle as well as to ascertain
oxygen uptake after taking BCAA showed a lower increase whether they had ever used muscle enhancers. Students who
in CK and LDH concentrations compared with people in the were found to have diseases or to have taken muscle enhancers
placebo group. Free fat acid (FFA), which is released from were excluded from the experiment. Before entering this
the adipose tissue and produces energy, is a major source experiment, the height, the weight, and the makeup of the
of energy during exercise that requires long-term endurance. subjects were measured to select the final 26 through the
The amount of FFA during exercise or after exercise is totally process to minimize errors. This was to minimize potential
affected by glycerol. Muscle capillaries are extended at the physical differences among the individual subjects in the
beginning of exercise to promote the use of FFA and this clinical trial. All the process also received approval by the
metabolic phenomenon ends at the end of the exercise. ethics committee. Thirteen participants were randomly
Glucose functions a basic role in carbohydrate metabolism assigned to the BCAA intake group and the others were
of skeletal muscle and is generated from muscle glycogen assigned to the placebo group. All the subjects were educated
and blood. In particular, glucose produced from the liver to understand the behavior that could affect the experiment
during exercise with intensity of about 60 percent of maximal and submitted the experimental agreement. Physical
oxygen uptake starts to decrease around 90 minutes after the characteristics of the subjects are shown in the <Table 1>.

Table 1. Physical characteristics of subjects

Item group Age (yrs) Height (cm) Weight (kg) Rest Heart rate (beat) BMI (kg/m2)
Experimental group (n = 13) 23.72 ± 2.34 171.69 ± 3.86 67.42 ± 8.56 65.01 ± 5.25 22.93 ± 4.28
Placebo group (n = 13) 22.45 ± 2.46 168.17 ± 3.39 66.21 ± 7.93 64.14 ± 7.06 23.40 ± 5.36
Values are Means ± SD.
 Dong-Hee Kim et al. / J Exerc Nutr Biochem 17(4):169-180, 2013 171

Experimental design BCAA ingestion

BCAA containing isoleucine (20%), valine (24%), and
In this study, the subjects rode on the bicycle ergometer
(Corival, Monark Inc., USA) and started gradual maximum leucine (46%) was dissolved in 500 ml of reverse osmosis
water and 80 mg/kg (Lee Han and others, 2002; Shimomura,
exercise after having taken BCAAs, which was dissolved in
et al., 2010) BCAA of weight was administered. The subjects
bottled water, and having drunken placebos (bottled water).
Dependent variables were measured five times; before drank reverse osmosis water as the placebos. Aspantam (15
mg/100 ml of reverse osmosis water) was added to both the
ingesting BCAAs and placebos; 10 minutes before exercise;
BCAA and the placebo so that the subjects could not
30 minutes into exercise; immediately after exercise; 30
minutes after exercise. Then the impact of the variables, which distinguish taste. As the double blind crossover method was
used, neither the subjects nor the researchers were able to
are related to fatigue substances, muscle damage substances,
know which liquid was the BCAA or the placebo. The
and energy metabolism substances, was analyzed. Preliminary
experiments were conducted to all the subjects one week subjects arrived at the laboratory in a fasting state and lied
down to rest for 30 minutes. Then they drank 500 ml of
before the experiment. Drinking, smoking, and caffeine intake
BCAAs or placebos 50 minutes before exercise in accordance
were banned three days before the experiment. All of the
research was conducted at the same time every day in order with studies (Leibetseder, et al., 2006) suggesting any longer
than this result in the bodily elimination of the BCAAs.
to minimize the effects of variables including the experimental
time, conditions, and the subjects’ circadian rhythm. Before
the experiment, the researchers randomly determined the order Taking blood samples
The subjects rested 48 hours before the experiment and
in advance and proceeded with every experiment in that order.
fasted 12 hours before the experiment. They were asked to
BCAAs and placebos were taken in accordance with a
double-blind trial. The experiment was performed with an arrive at the laboratory three hours before the experiment to
measure their heights and weights. This was followed by a
interval of one week to minimize the effects of previous exercise.
30-minute rest. Catheters were inserted into the veins in the
cubital fossas of the subjects who wore Polar (FIN-90440,
Experimental procedure and method Filand) watches. The experiment started 50 minutes after the
Exercise Method subjects took BCAAs or placebos. Ten milliliters of blood
Subjects fasted on the day of the experiment and got plenty was taken from the veins in the cubital fossa using a
of rest three hours before exercise. A week before the disposable syringe five times; before ingesting BCAAs or
experiment, each individual subject’s maximal exercise capacity placebos; 10 minutes before exercise; 30 minutes into exercise;
(maximal oxygen uptake, VO2max) was measured. The immediately after exercise; 30 minutes after exercise. The
Astrand-Rhyming protocol (1965) was used . The first starting analysis was conducted by The Clinical Pathology Center of
load was 600 kg·m/min (50 rpm, 2 kp; 100 w) and the load J University.
was increased by 300 kg·m/min (50 w) every two minutes.
For endurance test, the subjects started exercise with an Test items and methods
intensity of 1 kp (50 w, 50 rpm) and the intensity was
increased by 2 kp (100 w, 50 rpm) five minutes later. Ten The test methods to analyze fatigue substances include a
minutes later, the intensity (kp) was increased to 70 percent specific vehicle of EDTA-2Na with 8% Hypochlorite acid4
of individual maximal oxygen uptake (VO2max) and (HCIO4), 1% EDTA and 1% Ascorbic acid as a phosphorous
continued until the person could not work out any longer. acid at a pH of 3.5 in order to remove protein in Serotonin
Each individual’s point of exhaustion was determined as (5-HT). The study collected blood samples to agitate them
follows: when the subjects themselves said that they could in the centrifuge (HANIL Centrifuge MF_80) at a rate of
not continue exercising at the intensity corresponding to the 12,000 rpm for 20 minutes, and moved only the necessary
19th level of the Borg Scale (1982): when they were not able amounts of them to the sample injection tube to measure them
to maintain 50 rpm on the bicycle ergometer for five seconds based on a L-8200 type of HPL (high performance liquid
or longer: their heart rates were greater than the maximum chromatography). To measure ammonia, this study used a
heart rate (220-age) minus 10bpm (Heyward, 2010). During spectrophotometer (CL-750) to observe Berthelot responses,
the experiment, an average temperature of 20-24℃ was and employed the spectrophotometer to measure lactate in the
maintained in the laboratory and an average, relative humidity blood based on an enzyme method. To analyze CK, a muscle
of 40-60% was maintained. damage substance, the study used vacuum tubes for blood
172 BCAA intake during endurance exercise

test to collect blood samples, which were then stirred at rates Data processing
of 2500-3000 rpm for 15-20 minutes. The necessary amounts
The study measured the mean and standard deviation of
of bloods for measurement were once again extracted to
each variable by using the SPSS (ver 17.0). The study
analyze them with a dry biochemical analyzer (Kodak
conducted the repeated ANCOVA with ‘before the intake of
EKTACHEM DTSC Ⅱ, USA). To measure LDH, 2.70ml of
BCAA’ set up as a common variant to verify mean differences
Trisbuffer (57.5mmol/l), 0.1ml of NADH solution and 0.1ml
between groups and times. After having verified the gradients
extracted tissues were mixed evenly in a cuvette with a
of the data prior to the test, the study found that the gradients
diameter of 1cm, and were then warmed up in a water at
of the data were all identical. In addition, in case that a
30℃ for 10-20 minutes. The reactants was added with 0.2ml
significant difference happened, the study carried out a post
of pyruvate solution and evenly stirred before being measured
verification test based on the Duncan method. The analysis
with the dry bio-chemical analyzer (Kodak EKTACHEM
on the correlation between the variables within each subject
DTSC Ⅱ) equipped with a heating device at a rate of 340nm
group was done with the Pearson correlation method.
at intervals of half or 1 minute for 3-6 minutes. To determine
FFA, the study an energy metabolism substance, the study
collected blood samples with the vacuum tube equipped with
a clot activator designed to precipitate fibrinogen, agitated
STUDY RESULTS
them in a centrifuge at rates of 2,500-3,000 rpm for 15-20
minutes, added 50 µl STD solution to 0.5 ml extracted blood Changes in fatigue substances, muscle damage Substances
serums to mix them a SICDIA NEFAZYME test reagent, and and energy metabolism substances after endurance exercises
finally used Hitachi 7150 (Hitachi, Japan) to measure FFA
Changes in Fatigue Substances after Endurance Exercises
after putting them in a cold water for 5 minutes. To measure
The serotonin levels in the experimental group showed a
glucose, the study agitated the collected blood samples in a
tendency of decreasing at 10 min before exercise, 30 min
centrifuge at rates of 2,500-3,000 rpm for 15-20 minutes and
into exercise, immediately after exercise, and until 30 min
analyzed the supernatant liquid with a dry bio chemical
after exercise, in comparison with those before the intake of
analyzer (Kodak EKTACHEM DTSC Ⅱ, USA).
Table 2. The change of fatigue substances by the endurance exercises
10 min before 30 min into Immediately after 30 min after F
Item Group Berore ingesting post-hoc
exercisea exerciseb exercisec exercised Pr > F
serotonin EG 16.00 8.63 7.02 7.80 3.77
(µg/dl) 21.87 7.25 4.27 4.15 3.81 3.424
-
8.62 7.04 7.80 3.75 .324
MV ± SE
6.43 1.22 1.01 1.21
PG 15.77 19.93 9.45 12.40 10.95
8.52 30.02 6.22 2.54 5.81 4.104
-
19.95 9.43 12.40 10.97 .278
MV ± SE
6.43 1.22 1.01 1.21
ammonia EG 241.17 212.50 268.50 301.67 179.67
(umol/L) 57.64 35.98 42.42 72.48 56.56 9.09 A:C
217.41 275.25 304.08 182.36 .0001 B,C:D
MV ± SE
6.93 11.12 18.18 12.12
PG 269.00 169.00 218.00 248.83 165.67
113.69 42.55 69.41 52.02 27.02 5.621
A,B,D:C
164.10 211.25 246.42 162.97 .001
MV ± SE
6.93 11.21 18.18 12.12
lactate EG 13.43 10.60 47.62 14.88 13.18
(mmol/L) 8.57 1.33 13.42 2.47 2.44 54.251
B:A,C,D
10.59 47.39 14.88 13.10 .0001
MV ± SE
.82 4.70 2.77 .70
PG 9.65 12.08 47.92 31.32 16.12
1.79 3.59 17.40 12.67 2.21 32.026
A,D:BC
12.08 48.14 31.32 16.20 .0001
MV ± SE
.82 4.70 2.77 .70
EG, experimental group; PG, placebo group.
 Dong-Hee Kim et al. / J Exerc Nutr Biochem 17(4):169-180, 2013 173

the BCAA, while the ammonia levels tended to be increased dropped by a statistically significant degree at immediately
at10 min before exercise, 30 min into exercise and after exercise and 30 min after exercise. In regard to the
immediately after exercise, compared with those before the differences between the subject groups, the serotonin levels
intake of the BCAA, but showed a statistically significant in the experimental group were recorded lower at 10 min
drop at 30 min after exercise. The lactate levels showed a before exercise and 30 min after exercise than the placebo
statistically important increase at 30 min into exercise, group, while their ammonia levels were higher than the
compared with those before the intake of the BCAA, but placebo group at 10 min before exercise, 30 min into exercise
dropped by a statistically important degree at immediately and 30 min after exercise. The lactate levels in the
after exercise and 30 min after exercise. A statistically experimental group were lower than the placebo group at
significant difference in the interaction between the subject immediately after exercise. The results of the covariance
groups was observed at immediately after exercise. The analysis for the difference verification on changes in serotonin
serotonin levels in the placebo group were decreased at 10 showed a statistically important difference in the interaction
min before exercise and 30 min into exercise compared with between the subject groups, while there was no statistically
those before the intake of the BCAA, while their ammonia meaningful disparity in regard to the interactions among the
levels were increased at 10 min before exercise and 30 min times and between the groups and times <Table 3>. The
into exercise, in comparison with those before the intake of results of the covariance analysis for the difference
the BCAA, but dropped by a statistically important degree verification on changes in ammonia levels indicated a
at 30 min after exercise. The lactate levels showed a statistically important difference in the interactions between
statistically important increase at 30 min into exercise, the subject groups and among the times, while there was no
compared to those before the intake of the BCAA, but statistically meaningful disparity in the interaction between

Table 3. The result of repeated ANCOVA on the change of serotonin between groups and time
Source DF TypeⅢSS Mean Square F-Value Pr > F
covariance (pre-administration) 1 11.453 11.453 .090 .767
group 1 978.152 978.152 7.704 .011
error (group) 21 2666.228 126.963
time 3 432.495 144.165 1.037 .383
time×covariance (pre-administration) 3 471.951 157.317 1.131 .343
group×time 3 265.522 88.507 .636 .594
error (time) 63 8761.844 139.077

Table 4. The result of repeated ANCOVA on the change of ammonia between groups and time
Source DF Ⅲ
Type SS Mean Square F-Value Pr > F
covariance (pre-administration) 1 64832.093 64832.093 16.271 .001
group 1 55227.681 55227.681 13.861 .001
error (group) 21 83673.324 3984.444
time 3 25781.634 8593.878 6.964 .001
time×covariance (pre-administration) 3 11474.725 3824.908 3.100 .033
group×time 3 6983.682 2327.894 1.887 .141
error (time) 63 77739.525 1233.961

Table 5. The result of repeated ANCOVA on the change of lactate between groups and time
Source DF Ⅲ
Type SS Mean Square F-Value Pr > F
covariance (pre-administration) 1 6.194 6.194 .065 .802
group 1 645.923 645.923 6.735 .017
error (group) 21 2014.035 95.906
time 3 3822.763 1274.254 14.872 .001
time×covariance (pre-administration) 3 7.203 2.401 .028 .994
group×time 3 893.500 297.833 3.476 0.21
error (time) 63 5397.815 85.68
174 BCAA intake during endurance exercise

the groups and times <Table 4>. The results of the covariance exercise and 30 min after exercise. The LDH levels were
analysis for the difference verification on changes in lactate diminished at 10 min before exercise in comparison with those
levels showed that there were statistically important differences before the intake of the BCAA and started to increase at 30
in the interactions between the subject groups, among the min into exercise before falling again at immediately after
times, and between the groups and times <Table 4>. exercise and 30 min after exercise. The CK levels in the
placebo group were lowered compared with those before the
Change in muscle damage substances after endurance intake of the BCAA at 10 min before exercise but started
exercise to increase at 30 min into exercise before being dropped again
The CK levels in the experimental group were decreased at immediately after exercise and 30 min after exercise. Their
at 10 min before exercise compared with those before the LDH levels were decreased at 10 min before exercise in
intake of the BCAA but started to increase at 30 min into comparison with those before the intake of the BCAA and
exercise before being dropped again at immediately after started to increase at 30 min into exercise before falling again

Table 6. The change of muscle damage substances by the endurance exercise

10 min before 30 min into Immediately after 30 min after F
Item Group Berore ingesting post-hoc
exercisea exerciseb exercisec exercised Pr > F
CK EG 351.33 312.83 336.33 313.50 292.50
(U/L) 122.16 102.23 117.67 106.91 113.29 3.677
-
278.47 292.04 276.96 253.84 .161
MV ± SE
13.32 10.26 16.69 12.24
PG 251.00 251.50 271.00 253.33 235.33
84.16 59.07 73.84 76.91 57.30 4.448
-
285.87 315.29 289.87 273.99 .121
MV ± SE
13.32 10.26 16.69 12.24
LDH EG 1175.17 985.17 1115.17 770.17 748.67
(U/L) 754.89 363.62 629.88 370.51 238.69 16.612
B:C,D
982.24 1065.52 783.04 734.53 .0001
MV ± SE
125.93 132.69 136.02 90.79
PG 1002.83 1056.00 1266.17 949.67 942.33
483.60 477.88 519.06 547.97 389.39 16.612
B:C,D
1058.93 1315.82 936.79 956.47 .0001
MV ± SE
125.93 132.69 136.02 90.79

Table 7. The result of repeated ANCOVA on the change of CK between groups and time
Source DF Ⅲ
Type SS Mean Square F-Value Pr > F
covariance (pre-administration) 1 569156.025 569156.025 96.570 .001
group 1 4871.398 4871.398 .827 .374
error (group) 21 123767.809 5893.705
time 3 3175.882 1058.627 1.877 .143
time×covariance (pre-administration) 3 5240.271 1746.757 3.097 .033
group×time 3 735.651 245.217 .435 .729
error (time) 63 3538.562 564.104

Table 8. The result of repeated ANCOVA on the change of LDH between groups and time
Source DF Ⅲ
Type SS Mean Square F-Value Pr > F
covariance (pre-administration) 1 862916.144 862916.144 1.822 .191
group 1 726004.264 726004.264 1.533 .229
error (group) 21 9945873.689 473613.033
time 3 891650.255 297216.752 3.698 .016
time×covariance (pre-administration) 3 2517931.007 839310.336 10.419 .001
group×time 3 105798.690 35266.230 .438 .727
error (time) 63 5075125.826 80557.553
 Dong-Hee Kim et al. / J Exerc Nutr Biochem 17(4):169-180, 2013 175

at immediately after exercise and 30 min after exercise <Table Change in energy metabolism substances after endurance
6>. The results of the covariance analysis for the difference exercises
verification on changes in CK did not show any statistically The glucose levels in the experimental group were
important difference in the interactions between the subject decreased at 10 min before exercise, 30 min into exercise,
groups, among the times and between the groups and times at immediately after exercise and 30 min after exercise,
<Table 7>. The results of the covariance analysis for the compared to those before the BCAA intake, while their FFA
difference verification on LDH changes did not indicate any levels were dropped at 10 min before exercise, immediately
statistically meaningful disparity in the interactions between after exercise and 30 min after exercise, in comparison with
the subject groups, nor between the groups and times, while those before the intake of the BCAA. The glucose levels in
a statistically meaningful difference was observed in the the placebo group were diminished at 10 min before exercise
interaction among the times. and until the recovery 30 min during exercise compared to
those before intake and then started to increase again after

Table 9. The change of energy metabolism substances by the endurance exercises

10 min before 30 min into Immediately after 30 min after F
Item Group Berore ingesting post-hoc
exercisea exerciseb exercisec exercised Pr > F
Glucose EG 96.83 78.83 82.83 72.67 68.50
(mg/dl) 6.34 10.29 6.94 6.89 9.43 4.987
.063 -
78.98 80.71 71.47 67.42
MV ± SE
2.56 1.99 1.71 2.38
PG 89.33 88.67 86.50 76.67 78.67
4.70 3.75 6.68 3.60 4.54 5.084
-
88.52 88.62 77.87 79.74 .058
MV ± SE
2.56 1.99 1.72 2.38
FFA EG 642.17 395.67 415.17 1605.67 1582.50
(µEq/L) 390.04 204.72 266.78 548.03 766.65 19.713
A,B:C,D
322.37 331.05 1401.71 1328.15 .0001
MV ± SE
71.17 44.13 63.54 80.61
PG 352.17 479.17 433.33 1029.33 968.00
113.79 319.61 153.50 32.82 225.70 20.693
A,B:C,D
552.46 517.45 1233.29 1222.35 .0001
MV ± SE
71.17 44.13 63.54 80.61

Table 10. The result of repeated ANCOVA on the change of glucose between groups and time
Source DF Ⅲ
Type SS Mean Square F-Value Pr > F
covariance (pre-administration) 1 219.797 219.797 3.227 .087
group 1 1314.072 1314.072 19.293 .001
error (group) 21 1430.369 68.113
time 3 136.279 45.426 1.175 .326
time×covariance (pre-administration) 3 127.075 42.358 1.096 .357
group×time 3 77.345 25.782 .667 .575
error (time) 63 2434.758 38.647

Table 11. The result of repeated ANCOVA on the change of glucose between groups and time
Source DF Ⅲ
Type SS Mean Square F-Value Pr > F
covariance (pre-administration) 1 8188714.939 8188714.939 81.166 .001
group 1 23757.429 23757.429 .236 .632
error (group) 21 2117892.478 100852.023
time 3 825018.266 275006.089 9.817 .001
time×covariance (pre-administration) 3 2069691.575 689897.192 24.627 .001
group×time 3 573690.998 191230.333 6.826 .001
error (time) 63 1764842.008 28013.365
176 BCAA intake during endurance exercise

30 min after exercise, while their FFA levels were decreased relationship, while those between LDH and FFA and between
at 10 min before exercise and 30 min after exercise but started FFA and Glucose showed a negative relationship <Table 12>.
to increase again at immediately after exercise and 30 min
after exercise. In regard to the differences between the subject Correlation of fatigue substances, muscle damage substances,
groups, the glucose levels of the experimental group were energy metabolism in placebo group
lower than those of the placebo group at 30 min after exercise. The analysis results on the correlation among fatigue,
The results of the covariance analysis for the difference muscle damage and energy metabolism substances in the
verification on changes in glucose showed a statistically experimental group showed that the correlation between CK
important difference in the interaction between the subject and LDH showed a very strong positive relationship, while
groups, while the interaction among the times or between the that between FFA and glucose showed a very strong negative
groups and times did not indicated any statistically meaningful relationship <Table 13>.
disparity <Table 10>. The results of the covariance analysis
for the difference verification on FFA changes did not show
any statistically important difference in the interaction DISCUSSION
between the subject groups, while there was a statistically
meaningful difference in the interactions among the times and
Exercise and fatigue substances
between the groups and times <Table 11>.
The BCAA is the most easily oxidized type of amino acid
in the skeletal muscles and serves as an important energy
Analysis on correlation between fatigue substances, muscle
substrate when carbohydrate becomes exhausted after a
damage substances and energy metabolism substances
prolonged exercise, as the contribution rate of fat as an energy
Correlation between fatigue substances, muscle damage source increases [21]. Serotonin is known to be involved in
substances and energy metabolism substances body temperature, blood pressure, internal secretion, appetite,
According the analysis results on the Pearson correlation sexual behavior, exercise, aggression, and pains [10] and
coefficient among fatigue, muscle damage and energy adjust various biological functions such as memory and
metabolism substances in the experimental group, the cognition [1]. According to the previous studies on serotonin
correlations between LDH and ammonia, between CK and associated with exercise, the results of the measurement for
glucose and between glucose and serotonin showed a positive the activation level of serotonin at the raphe at the rate of

Table 12. Correlation of fatigue substances, muscle damage substances, energy metabolism substances in experimental group
CK LDH Ammonia FFA Lactate Glucose Serotonin
CK 1
LDH .119 1
Ammonia -.154 .466** 1
FFA -.095 -.371** -.215 1
Lactate -.044 .125 .189 -.224 1
Glucose .300* .079 -.042 -.451** .018 1
Serotonin -.113 -.216 .129 -.201 -.197 .411** 1
**p < 0.01

Table 13. Correlation of fatigue substances, muscle damage substances, energy metabolism in placebo group
CK LDH Ammonia FFA Lactate Glucose Serotonin
CK 1
LDH .591** 1
Ammonia .249 .167 1
FFA .217 -.157 .111 1
Lactate .128 .171 .219 .077 1
Glucose -.195 .072 -.164 -.635** -.213 1
Serotonin -.023 -.177 .233 .172 -.132 .177 1
**p < 0.01
 Dong-Hee Kim et al. / J Exerc Nutr Biochem 17(4):169-180, 2013 177

5-HIAA/5-HT showed that an animal group which worked endurance exercise rose due to the activation of fat
out up to 50% of the maximum exercise capacity showed oxidization, while that of the protein as an energy source was
a significantly higher activation percentage compared with the relatively dwindled, reducing the concentration of ammonia
other animal group which exercised up to the maximum in the blood. Lactate was formed as a result of anaerobic
exercise capacity [5]. Moreover, an increase in serotonin was metabolism of glucose during exercise, and is the important
reported in white rats after exercise, according to the study barometer in determining the limitation factors of muscle
on the effects of intensity of treadmill exercise and duration fatigue and muscle activity. In this study, the lactate levels
of mild-treadmill exercise on serotonin activity in the brain of the two groups, respectively, showed a statistically
of rats [11]. This study measured the levels of serotonin, significant increase at 30 min into exercise. The lactate levels
ammonia and lactate, with an aim to assess the effect of the in the BCAA intake group indicated a statistically meaningful
intake of BCAA on fatigue substances, and found that the decrease at immediately after exercise and returned to a stable
serotonin in a group that consumed BCAA was decreased level at 30 min after exercise. In regard to the variances
at 10 min before exercise and 30 min into exercise but slightly between the groups, the BCAA intake group showed a lower
increased at immediately after exercise before being dropped concentration of lactate at immediately after exercise than the
again at 30 min after exercise. In the placebo group, the level placebo group. The reason why the lactate level in the blood
of serotonin was increased at 10 min before exercise but reached the peak at the end of exercise is because the amounts
started to be decreased at 30 min into exercise being escalating of oxygen uptake and that oxygen which transported to the
again at immediately after exercise and 30 min after exercise. muscle tissues were reduced as the exercise proceeds, which
In regard to the variances between the subject groups, the in turn caused the accumulation of lactate in the body by
placebo group showed a statistically significant difference; it hindering pyruvate from entering the TCA cycle efficiently
indicated a higher serotonin concentration at 10 min before and transforming it into lactate.
exercise and 30 min after exercise than the BCAA intake
group, while the serotonin level in the BCAA intake group
Exercise and muscle damage substances
was lower than the placebo group at all test times, which
is identical to the results of the study by Lee Han, et al. The concentrations of CK and LDH, both of which play
[12]. The study results by Wagenmakers, et al. [20]. which an important role in adjusting the energy metabolism required
administered the BCAA into those patients who suffered from for muscle activity, can be used as a crucial indicator in
McArdle's disease and had them to ride a bicycle ergometer, determining the levels of muscle damage and body robustness
suggested that the ammonia concentration after exercise had [19]. Moreover, LDH, an indicator of muscle damage, is
increased by about 10 times compared to that in a stable known to play a crucial role in formation and transformation
condition. MacLean and Graham[15] administered the BCAA of lactate in muscle cells, among muscle activities. As an
into healthy people with a normal level of glycogen oxidization and reduction enzyme, it converts pyruvic acid
concentration and had them ride a bicycle ergometer. The into lactate in the process [17]. Liu, et al. [14] reported that
study results showed that the BCAA intake group indicated the CK and LDH levels in the group who consumed the
a significantly higher ammonia concentration in the blood BCAA for 5 weeks were measured after endurance and elastic
after exercise than the non-BCAA intake group. In this study, band exercises. Greer, et al. [7] reported that the BCAA intake
the ammonia levels in the both groups were increased at 10 group showed lower concentrations of CK and LDH after
min before exercise, 30 min into exercise and immediately they rode a bicycle ergometer at a rate of 50% of maximum
after exercise, but were decreased by a statistically significant oxygen uptake capacity for a prolonged time than otherwise.
degree at 30 min after exercise. In regard to the variances Matsumoto, et al. [16] reported that a group of 12 long
between the groups, the placebo group indicated a statistically distance runners who consumed beverages containing the
meaningful disparity with a higher concentration of ammonia BCAA while undergoing a three-day intensive training
at 30 min after exercise compared with that of the BCAA showed lower concentrations of both CK and LDH compared
intake group, while the BCAA intake group recorded a higher with the placebo group. The study indicated that the CK levels
concentration of ammonia at nearly all test times, which is of both groups were increased at 30 min into exercise but
similar to the results of the studies by MacLean and Graham reduced to lower than pre-exercise level at immediately after
[15]. The ammonia concentration was increased during exercise and 30 min after exercise. The CK concentration
exercise but was reduced to a stable level during recovery. change patterns depending on test times of both groups
It is presumed that the ratio of fat as an energy source during showed similar results, but the placebo group had a higher
178 BCAA intake during endurance exercise

concentration of CK than the BCAA intake group at all test during exercise but is preplaced with FFA at immediately after
times. The LDH levels of both groups were higher at 30 min exercise, for example, in case of more than one hour of
into exercise but were decreased at immediately after exercise exhaustive exercise. If all the above mentioned study results
and 30 min after exercise, compared with those before the are taken into consideration, the BCAA intake can lower the
intake of the BCAA. The LDH concentration change patterns concentration of serotonin, a central fatigue substance, which
depending on test times showed the same results. However, can in turn reduce the concentrations of muscle damage
the placebo group had a higher concentration of LDH than substances such as CK and LDH. That is to say that the less
the BCAA intake group. If all the above mentioned study secretion of fatigue substances means the less secretion of
results are taken into account, it is presumed that an reduction those enzymes that can cause muscle damages, and subsequently
of concentration of muscle damage substances such as CK can enhance exercise performance. Therefore, the intake of
and LDH can help contribute to enhancing exercise the BCAA is presumed to contribute to reducing the
performance. concentrations of muscle damage substances such as CK and
LDH and enhancing the exercise performance. Meanwhile,
an increase in the ammonia concentration can deteriorate
Exercise and energy metabolism substances
exercise performance, while a reduced secretion of FFA
Exercise can increase the influx of glucose into cytoplasm means a decreased secretion of LDH. In addition, the study
by rapidly stimulating the glucose transport activity in a confirmed that glucose is used as an energy source during
muscular fibrous coat [6]. The concentration levels of plasma exercise but is replaced with FFA at immediately after
glucose and FFA during exercise have a strong correlation. exercise, for example, in case of more than one hour of
Meanwhile, Gualano, et al. [8] reported that a group which exhaustive exercise, which in turn can improve exercise
consumed 300mg of BCAA per unit weight on a daily basis performance and plays an effective role in preventing muscle
showed a higher concentration of glucose after an exhaustive damages. However, as this study was based on the results
exercise. However, this study observed that the glucose levels of those previous studies which suggested that the intake of
in the BCAA intake group were slightly increased at 10 min the BCAA before exercise is the most effective in enhancing
before exercise and 30 min into exercise and were decreased exercise performance, more follow-up studies should be
at immediately after exercise and 30 min after exercise, in conducted in consideration of other variables such as gender,
comparison with those before the intake. When it comes to age, disease and nutrition status, and also more studies on
the variances between the groups, the placebo group showed the effect of the BCAA administration conditions (dosage,
a statistically meaningful disparity with a higher concentration duration, administration type) on exercise types, intensity and
of glucose at 30 min after exercise compared to the BCAA times need to be carried out in the future.
intake group. Although the glucose concentration change
patterns depending on test times showed similar results in
both of the groups, the placebo group showed a higher CONCLUSION
concentration of glucose at all test times than the BCAA
intake group. The concentration levels of FFA in the BCAA This study came to the following conclusions after
intake group showed a statistically meaningful increase at 10 analyzing the effects of the intake of BCAA on fatigue
min before exercise, 30 min into exercise, immediately after substances, muscle damage substances and energy metabolism
exercise and 30 min after exercise. The FFA concentration substances during endurance exercises.
change patterns depending on test times showed that the
BCAA intake group showed a lower concentration at 10 min 1. When it comes to fatigue substances, the placebo group
before exercise and 30 min into exercise, in comparison with showed a higher level of serotonin and lower levels of
the placebo group, but indicated a higher concentration of ammonia and lactate than the BCAA intake group.
FFA at immediately after exercise and 30 min after exercise 2. In regard to muscle fatigue substances, the placebo group
than the BCAA intake group. Moreover, the BCAA intake showed higher levels of CK and LDH at all test times than
group showed a higher concentration of FFA at immediately the BCAA intake group.
after exercise especially after one hour of exhaustive exercise, 3. When it comes to energy metabolism substances, the
which is identical to the results of the studies by Zhang, et placebo group showed a higher level of glucose than the
al. [22] and by Gualano, et al. [8]. It is presumed that this BCAA intake group, while the BCAA intake group showed
is due to the fact that the glucose is used as an energy source a higher increase in the FFA concentration than the BCAA
 Dong-Hee Kim et al. / J Exerc Nutr Biochem 17(4):169-180, 2013 179

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