Nutrients 2010, 2, 75-98; doi:10.

3390/nu2010075


nutrients
ISSN 2072-6643
www.mdpi.com/journal/nutrients
Review
Effects of Beta-Alanine on Muscle Carnosine and Exercise
Performance: A Review of the Current Literature

Julie Y. Culbertson
1,
*, Richard B. Kreider
1
, Mike Greenwood
2
and Matthew Cooke
3


1
Exercise and Sport Nutrition Laboratory, Department of Health and Kinesiology, Texas A&M
University, College Station, TX 77845, USA; E-Mail: [email protected]
2
Department of Health, Human Performance and Recreation, Baylor University, Waco, TX 73019,
USA; E-Mail: [email protected]
3
Schools of Medicine & Health Movement Studies, The University of Queensland, Herston,
Queensland, Australia; E-Mail: [email protected]

* Author to whom correspondence should be addressed; E-Mail: [email protected];
Tel.: +1 (979) 458-1484.

Received: 16 November 2009 / Accepted: 6 January 2010 / Published: 25 January 2010

Abstract: Muscle carnosine has been reported to serve as a physiological buffer, possess
antioxidant properties, influence enzyme regulation, and affect sarcoplasmic reticulum
calcium regulation. Beta-alanine (-ALA) is a non-essential amino acid. -ALA
supplementation (e.g., 26 grams/day) has been shown to increase carnosine
concentrations in skeletal muscle by 2080%. Several studies have reported that -ALA
supplementation can increase high-intensity intermittent exercise performance and/or
training adaptations. Although the specific mechanism remains to be determined, the
ergogenicity of -ALA has been most commonly attributed to an increased muscle
buffering capacity. More recently, researchers have investigated the effects of co-ingesting
-ALA with creatine monohydrate to determine whether there may be synergistic and/or
additive benefits. This paper overviews the theoretical rationale and potential ergogenic
value of -ALA supplementation with or without creatine as well as provides future
research recommendations.
Keywords: creatine monohydrate; anaerobic capacity; muscular fatigue; ergogenic aids


OPEN ACCESS
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1. Introduction

During moderate to high-intensity exercise, hydrogen ions (H+) begin to accumulate leading to a
drop in intramuscular pH and ultimately influencing muscle performance [1]. The greater the reliance
on glycolysis as the primary energy system (as seen with high-intensity exercise), the greater
production of lactic acid and H+, thus leading to further decreases in intramuscular pH. This decrease
in intramuscular pH has been suggested to be linked to fatigue-induced increases in muscle activation
and electromyographic (EMG) amplitude [2,3]. Thus, if the intramuscular pH decline can be prevented
or delayed, the fatigue induced EMG increase may also be delayed [4]. Beta-alanine (-ALA)
supplementation has been shown to increase muscle carnosine levels, which can act as a buffer to
reduce the acidity in the active muscles during high-intensity exercise [5-7]. -ALA supplementation
has been shown to have beneficial effects on exercise performance variables such as cycling
capacity [6], ventilatory threshold, and time to exhaustion [8]. For this reason -ALA has become a
widely used nutritional supplement for improving high-intensity exercise performance [4-6,9,10].
Creatine monohydrate supplementation has also been shown to have ergogenic effects by increasing
the availability of phosphocreatine (PCr), total creatine concentrations in the muscle, high intensity
exercise performance, and training adaptations [11]. For this reason, several studies have assessed
whether co-ingesting -ALA with creatine may have synergist and/or additive effects on exercise
capacity and/or training adaptations [4,10,12]. The purpose of this article is to review the theoretical
rationale and available scientific evidence assessing the potential ergogenic value of supplementing the
diet with -ALA with or without creatine. In addition, to discuss areas that future research should
address. This was accomplished by conducting a thorough review of the published literature related to
the physiological effects of carnosine and the role of -ALA and creatine supplementation on
carnosine levels, creatine levels, and exercise performance.

2. Carnosine

Carnosine (-alanyl-L-histidine) is a naturally-occurring histidine-containing compound found in
many animal tissues, including skeletal muscle, which is the most abundant source. Carnosine is a
multifunctional dipeptide with many roles including buffering [13,14], fighting free radicals [15,16],
enzyme regulation [17] and sarcoplasmic reticulum calcium (Ca
2+
) regulation [18,19]. Carnosine is
broken down in the body by carnosinase, which is found in most tissues except skeletal muscle,
partially explaining why carnosine concentrations are highest in this tissue [19]. Figure 1 shows the
chemical structure of carnosine.
Carnosine in human skeletal muscle generally ranges between 5-10 mM wet weight or
1540 mmol/kg dry weight [5]. Concentrations differ among animal species, in part due to the
differences in muscle mass [20]. For example, horses have been reported to have higher carnosine
concentrations than greyhound dogs [21]. Carnosine levels are typically higher in fast-twitch muscle
fibers compared to slow-twitch, which corresponds to the observation that animals exposed to frequent
sprints, explosive flight behaviors and prolonged hypoxic dives have higher initial carnosine
concentrations [5,21,22]. Human athletes involved in anaerobic sports such as sprinters [23,24] and
bodybuilders [25] have also been found to have higher intramuscular concentrations of carnosine.
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77
Exercise training has been reported to increase resting muscle carnosine concentrations in these athlete
types. For example, Gardner and colleagues [26] reported that exercise training increased the plasma
carnosinase activity and decreased carnosine excretion leading to greater muscle carnosine
concentrations [26]. Moreover, Suzuki and colleagues [27] examined the effects of sprint training on
muscle carnosine concentrations. Six male subjects performed sprint training twice a week for a total
of 16 training sessions. Each session involved either single (for weeks one and two) or a double (for
weeks three through eight) bout of 30 seconds of maximal sprinting on a cycle ergometer with 20
minutes of rest between sprints on the double bout days. Muscle samples were collected from the
vastus lateralis one week before training and again two days following the training protocol. Results
revealed that muscle carnosine content and mean power output significantly increased after the eight
weeks of training [27]. Tallon and coworkers [25] suggested the greater muscle carnosine content in
bodybuilders may be due to the chronic exposure to lower pH environments due to their training,
differences in their diet such as increased protein intake where carnosine can be found,
supplementation use, and/or possible anabolic androgenic steroid use [25].
Figure 1. Chemical structure of carnosine.


Carnosine was first discovered as an intracellular pH buffer in 1953 by Severin and colleagues [28]
using frog muscle tissue. Subsequent studies examining this relationship in human muscle tissue
followed thereafter [13,14,29-32]. When skeletal muscles are involved in moderate to intense exercise,
there is typically a generation of lactic acid and subsequent dissociation into lactate and H+, which can
alter the pH levels. It had previously been reported that the majority of protons produced during
exercise in the blood were buffered by the bicarbonate buffering system [33]. The pKa of this system
is 6.1, which is less than that of carnosine (pKa of 6.83), and thus a greater pH change is needed to
elicit benefits from this system. Since the pKa of carnosine is closer to the physiological pH, it is likely
that this is utilized sooner as a buffer during high-intensity exercise [1]. The imidazole group on the
histidine containing molecules, such as carnosine, makes it especially effective as a buffer. This group
has a pKa value close to that of the intracellular pH, therefore one of the nitrogens from the imidazole
ring can be used to accept a proton [34].
Early studies examined the role of carnosine in animal models. One study, utilizing chromatography
methodology to analyze rabbit and pigeon muscle samples, reported muscle dipeptides (mainly
carnosine and anserine) accounted for approximately 40% of the pH buffering capability in skeletal
muscle [13]. Later, Bump and colleagues [35] examined the carnosine concentrations in different
breeds of horses. They compared quarter horses (QH), thoroughbreds (TB) and standardbreds (SB) in
order to correlate buffering capabilities of the muscle to fiber type composition. The QH demonstrated
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78
less slow-twitch muscle fibers, greater fast-twitch glycolytic fibers, and fewer fast-twitch oxidative
muscle fibers compared to the other horses. Results showed QH had significantly greater amounts of
carnosine in their muscle. The researchers reported a positive correlation between carnosine
concentrations and fast-twitch glycolytic fibers and a negative correlation between carnosine and fast-
twitch oxidative fibers. The investigators inferred that intramuscular carnosine acted as an intracellular
buffer, although this was not directly measured. A later study conducted by Sewell and associates [36]
specifically examined the buffering capability of carnosine in different fiber types of horses. These
researchers found that carnosine contributed about 20% of the buffering in type I fibers and up to 46%
in Type IIb fibers. These findings are consistent with the findings that less lactic acid is accumulated in
Type I fibers due to the lower intensity muscle activity involved with this fiber type.
An early study by Kraemer and associates of humans utilizing carnosine supplementation [37]
reported no effect on the acid-base status or exercise performance using four subsequent 30 second
Wingate tests with only two minutes of rest between exercise bouts. In this regard, the researchers
evaluated ten trained and ten untrained males who consumed a total of 15 capsules of a supplement
containing 1000 mg dibasic sodium phosphate, 204 mg potassium bicarbonate and 12.5 mg L-
carnosine over a 3.5 day period. Placebo capsules were matched in sodium and potassium content.
Blood samples were taken at baseline prior to any exercise, immediately after each Wingate test, and at
three minutes after all exercise was completed. Though intramuscular carnosine levels were not
measured, the authors suggested that the amount of carnosine provided to subjects (about 185 mg) may
have been too low to have an impact on intramuscular carnosine levels [26,38,39] particularly since
previous animal studies had shown increases following a daily dose between 50-200 mg/kg of body
weight [38,40].
Human studies have shown that lowered pH levels can also negatively affect the excitation-
contraction coupling in the skeletal muscle [38,41]. The buffer efficacy in human muscle was
examined by calculating the buffering ability over the physiological pH range of 7.16.5. This study
involved 50 healthy active individuals who underwent a muscle biopsy from the lateral portion of the
quadriceps femoris muscle. Anserine and carnosine were analyzed in neutralized perchloric acid
extracts using high performance liquid chromatography (HPLC) methods. The Henderson-Hasselbach
equation was then used to indirectly calculate the buffer contribution across the pH range of 7.1 to 6.5.
It was estimated that carnosine was able to buffer between 2.4 and 10.1 mmol H
+
kg
-1
dry mass, which
corresponded to about 7% of the total muscle buffering [9]. Therefore, these results indicated that
carnosine played a minimal role in buffering pH.
Suzuki and coworkers [42] examined the effects of the non-bicarbonate buffers carnosine and
anserine. They had eight active males supplement with either a placebo or chicken breast extract
(CBEX) soup that contained 1.5 g carnosine and anserine. Subjects then performed ten sets of five
second maximal cycle sprints at 7.5% of their body weight as resistance. Blood samples were collected
at rest, one minute before exercise, after each exercise set, and immediately after the intervals to
measure blood-gas parameters, blood lactate and concentrations of carnosine and anserine. The
researchers found that supplementing the diet with CBEX delayed the decrease in bicarbonate during
intense exercise, but did not improve performance. These results support the initial use of carnosine as
a buffer instead of the bicarbonate system [42].
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79
Early studies with carnosine supplementation noted plasma carnosine levels failed to elevate due to
the high activity of carnosinase [26]. The researchers were able to measure only 14% of the ingested
carnosine in urine suggesting this was due to absorption in the gastrointestinal tract [26]. Later,
research pointed towards supplementing with -ALA and L-histidine instead to raise carnosine levels
since these are the precursors to carnosine. Dunnett and Harris [14] discovered that -ALA was able to
increase carnosine in muscle tissue. In their study, they supplemented horses with both -ALA and L-
histidine and found -ALA to have an additive response suggested to be due to the increase -amino
acid transport across the gastrointestinal tract. This was not observed for L-histidine, thus speaking to
the efficacy of -ALA instead to increase carnosine levels [14]. However, Tamaki and coworkers [43]
were able to show an increase in carnosine with histidine in rats [43].
Aside from the buffering effects, carnosine has shown to have other physiological roles, including
that of an effective antioxidant against oxidative stress [44]. Reactive oxygen species (ROS) can arise
from exercise in several proposed mechanisms including: an increase flow of electrons in the electron
transport system from increased respiration [45] or a decrease in pH can lead to oxygen being released
from hemoglobin and a subsequent increase in pO
2
in the tissues [46]. Some believe the development
of ROS to be related to muscle fatigue during activity [47,48].
Carnosine is also linked to enzyme regulation related to activation of myosin ATPase, which is used
to help maintain ATP stores [49]. Finally, carnosine has been noted to have a role in electron-
contraction (E-C) coupling in skeletal muscle. An early study by Lamont and Miller [50] showed 15
mM of carnosine resulted in a significant increase in Ca
2+
sensitivity in muscle fibers of Rana
temporaria [50]. More recently, Dutka and Lamb [51] examined if carnosine affects E-C coupling in
functional fibers under physiological conditions. They used mechanically skinned rat extensor
digitorum longus muscle fibers. Their results showed that carnosine did not affect Ca
2+
release from
the sarcoplasmic reticulum; however, carnosine was able to increase the Ca
2+
sensitivity of the
contractile components of the muscle fibers. Authors suggested the assistance in Ca
2+
sensitivity could
help maintain force production in the later stages of fatigue once Ca
2+
release begins to decrease.
Therefore, higher levels of carnosine can help offset the decrease in Ca
2+
as well as the accumulation
of H
+
ions during high-intensity exercise [51].
Since carnosine has a number of physiological roles, there are a number of future research
opportunities. Specifically, the exact mechanism of carnosine in its role to improve exercise
performance and/or reduce muscular fatigue needs to be studied. It will also be important to examine
how different nutritional strategies to increase carnosine levels in the muscle may optimize
physiological activity and/or exercise capacity.

3. Beta-Alanine

-ALA is a naturally occurring amino acid that is one of the precursors to carnosine, along with L-
histidine. Carnosine synthetase is the enzyme used to synthesize carnosine from -ALA and L-
histidine. -ALA is also likely to be the rate-limiting step in the synthesis of carnosine [14,52,53].
Carnosinase is the enzyme present in cells and serum that breaks down carnosine into -ALA and
L-histidine [54].
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80
-ALA supplementation in doses greater than 10 mg/kg of body weight has shown to cause a short
period of paraesthesia with increasing severity as the dose increases. However, when a large dose
around 40 mg/kg of body weight is ingested with CBEX, the paraesthesia did not occur. It is
hypothesized that this side effect is a result of the rapid high peak blood plasma concentrations of -
ALA with supplementation alone, since it is not experienced when -ALA is ingested through the diet
with histidine containing dipeptides such as carnosine in meat products [7].

3.1. Beta-Alanine and Carnosine

As previously mentioned, -ALA supplementation has recently been shown to significantly increase
intramuscular carnosine levels, which then corresponds to improvements in exercise performance [8].
Harris and colleagues [7] examined the effects of -ALA supplementation on human skeletal muscle
carnosine concentration in a series of studies. In one study, investigators examined the effects of four
weeks of -ALA or carnosine supplementation on muscle carnosine concentrations. The
supplementation protocol included consuming 800 mg of -ALA four times a day (i.e., 2.3 g/day) for a
total intake of approximately 90 g over the four week period (group I) or increasing doses of -ALA
through the supplementation period (average 6.4 g/day) for a total intake of about 146 g over the four
week period (group II). The carnosine supplementation group involved consuming increasing doses of
L-carnosine through the supplementation period for a total intake of 364 g of L-carnosine over the four
week period which corresponded to an intake of about 143 g of -ALA. A final group supplemented
with maltodextrin as a placebo in the same frequency as the -ALA and L-carnosine supplementation
groups. A muscle biopsy was taken before and after supplementation. Results revealed that each
supplement group showed significant increases in carnosine content. Mean carnosine content increase
(measured in mmolkg
-1
dm) was greatest with L-carnosine and was followed by groups II and I of -
ALA with values of 16.37 3.03 (p < 0.05), 11.04 2.68 (p < 0.05) and 7.80 0.36 (p < 0.05)
mmolkg
-1
dm, respectively. There was no change in the placebo group (1.87 1.73, p>0.05 mmolkg
-
1
dm). This corresponded to percent changes of 66%, 64%, 42%, and 10% for L-carnosine, group II,
group I and placebo group, respectively. They also indirectly calculated the contribution of carnosine
to buffering capacity between pH levels of 7.1 and 6.5 using the Henderson-Hasselbach equation. They
found that after four weeks of supplementation, carnosine accounted for about 14.2%, 14.3%, and
12.6% of the total muscle buffering capacity in the L-carnosine group, group II, and group I,
respectively [7].
Studies have also suggested that there does not appear to be an upper limit on increasing muscle
carnosine concentrations. For example, Derave and colleagues [5] supplemented trained male sprinters
with -ALA or placebo (maltodextrin) for four to five weeks. The supplementation protocol included
six daily doses of 400 mg capsules of either -ALA or maltodextrin totaling 2.4 g/day for the first four
days, 3.6 g/day for the next four days, and 4.8 g/day for the duration of the study. Interestingly, muscle
carnosine levels were increased even in individuals with high resting muscle carnosine
concentrations [5].
While -ALA and carnosine supplementation have been reported to increase muscle carnosine
levels, less is known about the time course of carnosine degradation. Carnosinase is responsible for the
hydrolyzation of carnosine and is mainly present in human plasma, which is why carnosine levels are
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81
much lower in the blood than in skeletal muscle, where this enzyme is not present [26]. -ALA
supplementation in doses of 4-6 g/day over time has been shown to increase carnosine by 20-30% after
two weeks, by 40-60% after four weeks, and up to 80% by ten weeks [6,55]. A study by Baguet and
colleagues [56] sought to determine the loading phase of carnosine and the time course of removal.
They included 20 males who supplemented with either -ALA or maltodextrin as a placebo for five to
six weeks. The investigators provided doses of 2.4 g/day for days one and two, 3.6 g/day for days three
and four, and 4.8 g/day for the remainder of the study duration. Using a proton magnetic resonance
spectroscopy (MRS), they measured the carnosine content in three different muscles (soleus, tibialis
anterior, and gastrocnemius) at four time points (pre-supplementation, during the last week of
supplementation and at weeks three and nine following the cessation of supplementation). They
determined that carnosine elimination occurs relatively slowly and in a linear pattern at an average rate
of 0.21 mM/week in both type I and II fibers. Authors suggest the slow clearance of carnosine is
indicative of the high stability of the metabolite [56]. Table 1 provides a summary of recent studies
examining the effects of -ALA supplementation on carnosine concentrations.
Table 1. Summary of the effects of -ALA supplementation on muscle carnosine concentrations.
Authors Population Supplementation Protocol Muscle Carnosine
Concentration Effects
Performance
Results
Baguet et al.,
2009 [56]
20 physically
active males
5-6 weeks of -ALA or
placebo (maltodextrin)
2.4 g/day first 2 days
3.6 g/day days 3-4
4.8 g/day to end of study


In soleus, carnosine increased
30% (p=0.003) with -ALA
and remained stable with
placebo (p=0.867)
In tibialis anterior, carnosine
increased 27% (p=0.005) with
-ALA and decreased 17%
(p=0.05) with placebo
In gastrocnemius, carnosine
increased 23% (p=0.038) and
did not change with placebo
(p=0.740).
Carnosine elimination was
measured at 3 and 9 weeks
after supplementation period
At 3 weeks, only 26.1% (in the
soleus), 20.1% (in tibialis
anterior) and 44.7% (in the
gastrocnemius) of the increase
had disappeared. There was
no difference between -ALA
and placebo at this point
(p=0.431)
At 9 weeks, carnosine levels in
all 3 muscles returned to
initial values
None
measured
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82
Table 1. Cont.
Harris et al.,
2006 [7]
Study 3:
21 physically
active males
Ages 26.1 5.6
yrs
4 weeks, 4 groups (I IV):
I) 800mg -ALA x 4
daily (avg. 3.2g daily
and 89.6g 4wk total)
II) 8 daily doses of
either 400 or 800mg
-ALA (avg. 6.4g
daily and 145.6g 4wk
total)
III) 8 daily doses of
1000 or 2000 mg L-
carnosine (364g 4wk
total L-carnosine,
corresponding to
143.3g -ALA)
IV) Placebo of
maltodextrin at
doses to match
groups II and III

Increase in carnosine
concentration greatest with
carnosine supplementation,
followed by group II, then
group II -ALA protocols.
Mean increase over 4 weeks
(mmolkg
-1
dm)
I) 7.80 .36 (p < .05)
II) 11.04 2.68 (p < .05)
III) 16.37 3.03 (p < .05)
IV) 1.87 1.73 (p>.05)
None measured
Derave et al.,
2007 [5]
15 male track
athletes
(sprinters) 18-24
yrs
4-5 weeks -ALA or
placebo (maltodextrin)
2.4g/day first 4 days
3.6g/day days 5-8
4.8g/day to end of study

Soleus:
47% with -ALA
No change with placebo
Gastrocnemius:
37% with -ALA
No change with placebo

No difference
between groups
for 400m running
performance
Hill et al.,
2007 [6]
25 physically
active males
10 weeks
-ALA:
4g/day wk 1
4.8g/day wk 2
5.6g/day wk 3
6.4g/day wk 4-10
-ALA group, from 19.0 to
30.1 mmol/kg (58.8%) at 4
weeks and up to 34.7
mmol/kg (80.1%) at 10 weeks
Not significant between weeks
4 and 10
No effect on
body mass
cycling
capacity time
at 110% with
-ALA

3.2. Beta-Alanine and Exercise Performance

Increases in muscle carnosine due to -ALA supplementation have resulted in significant effects on
several variables related to exercise performance. Some of these include improved time to fatigue on a
maximal cycle test [6], increased ability to sustain power output in the final ten seconds of the Wingate
test [31], delayed onset of neuromuscular fatigue during incremental cycle ergometry tests as noted by
increased physical working capacity (PWC
FT
), increased ventilatory threshold (VT) and time to
exhaustion (TTE) [8], and improvements in muscle torque during repeated bouts of intense dynamic
contractions [5].
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Since studies have reported that muscle carnosine levels are typically higher in fast-twitch muscle
fibers, which are most predominantly used in high-intensity anaerobic exercise bouts, it has been
hypothesized that -ALA supplementation could aid in anaerobic performance. In 2002, Suzuki and
colleagues [31] performed a study that did not involve any nutritional supplementation, but simply
analyzed muscle biopsy samples from the vastus lateralis before and after a 30-second maximal cycle
sprint Wingate test. The muscle samples were analyzed for carnosine content. Analysis showed a
direct relationship between carnosine concentration in skeletal muscle and performance on the 30-
second Wingate exercise test. This relationship lends itself to the question of efficacy of -ALA
supplementation in further improving anaerobic exercise performance.
Hill and coworkers [6] examined the effects of four and ten weeks of -ALA supplementation on
muscle carnosine concentration and high-intensity cycling capacity. They also sought to discover
whether the effects were muscle type specific. Physically active males supplemented with either -
ALA or maltodextrin as a placebo. -ALA was given in eight doses per day with increasing dose
amounts during the first four weeks ranging from 250-750 mg per dose. Subjects underwent muscle
biopsies and maximal cycle performance tests at various points during the study. The group
supplementing with -ALA had significantly greater muscle carnosine concentrations at four and ten
weeks from 19.9 1.9 to 30.1 2.3 (30.4%) and 34.7 3.7 (35.1%) mmolkg
-1
dm. There was no
significant change with placebo. The change between four and ten weeks with -ALA was not
significant despite the small increase (p~0.07). The results also indicated no difference between fiber
types, in that each showed similar increases in carnosine as measured by HPLC with fluorescence
detection. The authors suggested that the possible benefits from -ALA supplementation may be
limited to four weeks, which is in agreement with previous findings by Suzuki and coworkers [31] who
showed an increase in the ability to sustain power output after four weeks of supplementation with no
additional benefits observed at ten weeks [31].
Limited research has examined the effects of -ALA on sport-specific anaerobic performances.
Derave and colleagues [5] studied the effects of a four week supplementation period on athletic
performance, using a 400 m running race time trial. The researchers found no significant differences in
performance after supplementation, but suggested this may have been due to the short time period of
supplementation since it takes several weeks to induce carnosine loading. Using a proton MRS to
detect muscle carnosine concentrations, investigators showed an increase in carnosine concentrations
of 47% in the soleus muscle after -ALA supplementation with no significant increase after placebo
supplementation (8%). Both groups showed significant increases in carnosine concentrations in the
gastrocnemius but subjects supplementing their diet with -ALA observed a greater increase (37%
versus 16%) [5]. This is in contrast to the previously discussed study that reported performance
improvements after four weeks of supplementation [6]. The researchers suggested that this may be due
to the possibility that in trained athletes, a 400 m running performance is not necessarily limited by the
intracellular pH decrease, and therefore the buffering capabilities of the increased carnosine
concentrations would not be as critical of a component [5].
Another recent study sought to determine whether -ALA supplementation would affect endurance
cycling performance. Van Thienen and colleagues [57] evaluated whether -ALA supplementation
would enhance the final sprint performance during endurance cycling since many competitions are
won in the final seconds of the race after an all-out sprint. They studied 21 trained males who
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84
supplemented their diet with either -ALA or a maltodextrin placebo for eight weeks. The dose
gradually increased from 2 g/day for the first two weeks, 3 g/day for weeks three and four, to 4 g/day
for weeks five to eight. The exercise test involved 110 minutes of cycling in ten minute stages with
increasing intensity between 5090%. Following this, the subjects performed a 30 second all-out
sprint. The researchers reported that -ALA supplementation increased sprint peak power after a two
hour endurance exercise bout by 1115% (p=0.0001) and mean power output by 58%
(p = 0.005) [57].
In contrast to trained individuals, Smith and colleagues [58] recently examined the combined effects
of six weeks of -ALA supplementation and high-intensity interval training on endurance performance
in recreationally active males. In this study, 46 participants were randomly assigned to either -ALA
or placebo supplementation groups. Both groups trained at 90110% of their peak oxygen utilization
(VO
2
peak) for the first three weeks, followed by three weeks of training at 115% VO
2
peak. During the
training, they continually supplemented with 6 g/day of -ALA or a dextrose placebo for the first three
weeks and 3 g/day for the second three weeks. They showed increases in both groups for VO
2
peak,
time to reach VO
2
peak, and total work done. However, the group ingesting -ALA observed a greater
increase in VO
2
peak and time to reach VO
2
peak during the second three weeks of the training protocol
(p < 0.05), with no change in the placebo group. They also noted a significant increase in lean body
mass for the -ALA group after the first three weeks. These results suggest that -ALA
supplementation may enhance the effects of high-intensity interval training and improve endurance
performance in untrained individuals. Additionally, Smith and colleagues [59] examined the effects of
the same high-intensity interval training and -ALA supplementation protocol described above on
neuromuscular fatigue and function. The researchers reported that three weeks of the interval training
was sufficient to result in a significant increase in the EMG fatigue threshold (EMG
FT
). However, -
ALA supplementation did not promote greater benefits [59]. Table 2 presents a summary of recent
studies examining the effect of -ALA supplementation and carnosine loading on
exercise performance.
Table 2. Summary of recent -ALA supplementation and exercise performance studies.
Authors Population Supplementation
Protocol
Exercise Testing
Protocol
Performance Results
Baguet et
al., 2009
[60]
14 physically
active males
4 weeks of -ALA or
placebo
(maltodextrin)
2.4 g/day first 2
days
3.6 g/day days 3-4
4.8 g/day to end of
study

Maximal ramp exercise
test on cycle ergometer
to determine VO
2
peak,
VT and gas exchange
threshold

Pre and Post
supplementation: 3 x
6min cycle exercise
bouts at 50% power
output
Exercise-induced acidosis was
19% lower with -ALA
No difference in VO
2
throughout
exercise before or after
supplementation in either group
Time delay in the fast component
was significantly shorter with -
ALA than placebo
Does not support a role for
acidosis in O
2
deficit or the slow
component of VO
2
kinetics

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Table 2. Cont.
Stout et al.,
2006 [4]
51 males 4 groups:
Placebo 34 g
dextrose
Creatine 5.25 g
creatine monohydrate
and 34 g dextrose
-ALA 1.6 g -ALA
plus 34 g dextrose
-ALA+Creatine
5.35 g creatine
monohydrate, 1.6 g -
ALA and 34 g
dextrose
28 days of
supplementation:
4 doses/day - days 1-6
2 doses/day - days 7-
28

PWC
FT
test with EMG
measurements on a cycle
ergometer
-ALA may delay the onset
of neuromuscular fatigue,
but no additive effects of
creatine
Significant increase in
PWCFT with -ALA
(14.5%) and creatine plus -
ALA (11%) compared to
placebo
Stout et al.,
2007 [8]
22 females
Ages:
28.9 8.1
yrs (-ALA)
25.8 4.0
yrs (placebo)
4 weeks -ALA or
placebo
4 divided doses/day for
totals of:
3.2g/day days 1-7
6.4g/day days 8-28

Continuous graded
exercise test on cycle
ergometer for VO
2max
,
ventilatory threshold ,
PWC
FT
and TTE
-ALA delays onset of NMF
during incremental cycle
ergometry ( PWC
FT
, VT,
TTE)
Stout et al.,
2008 [61]
26 elderly
males and
females
90 days supplementation
with -ALA or placebo
(microcrystalline
cellulose)
3 doses/day of:
2.4 g -ALA or
2.4 g placebo

Continuous graded
exercise test on cycle
ergometer for PWC
FT

with EMG measurements
28,5% increase in PWC
FT

after 90 days of -ALA
Sweeney et
al., 2009
[62]
19 physically
active
college-aged
males
5 weeks -ALA or placebo
(rice flour)
4g/day week 1
6g/day weeks 2-5
2 sets of 5x5-sec sprints
with 45- sec recovery
between sprints and 2
min between sets
performed on non-
motorized treadmill at
15% body weight as
resistance
No between group
difference for peak or mean
horizontal power
No difference in % fatigue
No difference in blood
lactate pre- and post-testing
between groups

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86
Table 2. Cont.
Van Thienen
et al., 2009
[57]
17 healthy
young males
8 weeks -ALA or
placebo (maltodextrin)
2 g/day weeks 1-2
3 g/day weeks 3-4
4 g/day weeks 5-8
Simulated road race of
110 minutes intermittent
endurance with intensity
between 50% and 90% of
the maximal lactate
steady state (MLSS) in 10
minute stages.
Immediately after this,
they started a 10 minute
time trial at 100% MLSS
with voluntary increase of
intensity at each minute.

-ALA enhanced sprint
power output at the end of
the endurance race
compared to placebo
Zoeller et
al., 2007
[10]
55 males
ages 24.5
5.3 yrs
4 weeks, 4 groups (4
doses/day for first 6 days,
then 2 doses/day
Placebo 34g dextrose
Creatine 5.25g
creatine monohydrate
and 34g dextrose
-ALA 1.6g -alanine
and 34g dextrose
-ALA plus Creatine
5.25g creatine
monohydrate, 1.6g -
ALA and 34g dextrose

Continuous graded
exercise test on cycle
ergometer
in 5 cardio-respiratory
endurance variables with
creatine + -ALA
Combined supplementation
may delay the onset of VT
and lactate threshold during
incremental cycle exercise

3.3. Beta-Alanine and Exercise Training

Many athletes incorporate resistance exercise as part of their training. Resistance-exercise has been
reported to lower pH levels to around 6.8 during an exercise session [63,64]. Thus, -ALA
supplementation may provide ergogenic value to athletes engaged in resistance training due to the
heavy reliance on glycolytic systems in the exercises [9]. Several recent studies have examined this
hypothesis. For example, Kendrick and coworkers [9] examined the effects of ten weeks of resistance
training with and without -ALA supplementation on muscle carnosine concentration and performance
measures. Subjects consumed 6.4 g/day of -ALA or a maltodextrin placebo for ten weeks. Results
revealed that -ALA supplementation increased muscle carnosine levels by 12.8 8 mmol/kg dry
muscle weight in with previous research [6,7]. However, the researchers reported that -ALA
supplementation had no effects on whole body strength, isokinetic force production, muscular
endurance, or body composition [9].
In a follow-up study, Kendrick and colleagues [65] performed a study examining the effects of four
weeks of -ALA supplementation on isokinetic training adaptations and muscle carnosine content in
type I and II fibers. Fourteen male subjects were divided into two supplementation groups. Subjects
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87
ingested 800 mg of -ALA or a maltodextrin placebo eight times per day for four weeks (6.4 g/day).
Subjects trained three times a week for the first two weeks and four times a week for weeks three and
four. Each session consisted of ten sets of ten maximal 90 knee extension and flexion contractions at
180/sec on the right leg using a Kin-Com isokinetic dynamometer with one minute of rest between
sets. The left leg acted as the untrained control. Muscle biopsies were obtained from the trained and
untrained legs prior to and following the training and supplementation period. Results revealed that
carnosine content was increased in the trained (9.6 3.9 mmol/kg dry muscle) and untrained legs
(6.6 2.4 mmol/kg dry muscle) with no significant differences observed between groups. In addition,
no significant differences were observed between carnosine concentrations in type I and type II fiber
types. The researchers concluded that four weeks of isokinetic training is not effective in increasing
carnosine content and that -ALA supplementation serves to increase muscle carnosine concentration
in both untrained and trained type I and type II muscle fibers [65]. Other recent studies support
contentions that -ALA supplementation can enhance training adaptations [12,58,59]. Table 3 provides
a summary of recent studies on -ALA supplementation and exercise training.
Table 3. Summary of recent -ALA supplementation and exercise training studies.
Authors Population Supplementation
Protocol
Exercise Protocol Muscle Carnosine
Concentration Effects
Performance
Results
Hoffman
et al., 2006
[12]
33 male
strength
power
athletes
10 weeks
Creatine -
ALA (CA)
10.5g/day
creatine
monohydrate
and 3.2g/day -
ALA
Creatine (C)
10.5g/day
Placebo (P)
10.5g/day
dextrose
Resistance training program
4 days/week for 10 weeks
Not measured fatigue rate
in CA
lean
body mass
and % body
fat
No change in
power
measures
training
volume in
CA
Kendrick
et al., 2008
[9]
26 healthy
males, 19-
24 yrs
800mg x 8/day
for 4 weeks of -
ALA or placebo
(maltodextrin)
Resistance training
4days/wk for 10 weeks
-ALA 23.96
5.94 to 36.77 8.26
(p < 0.0001)
Placebo 29.17
9.82 to 27.29 9.52
(p > 0.05)
No difference in
whole body
strength or
isokinetic force

Kendrick
et al., 2009
[65]
14
Vietnamese
college
aged
students
4 weeks -ALA
or placebo
(maltodextrin)
800mg x 8/day
Single legged isokinetic
training
3 sessions weeks 1-2
4 sessions weeks 3-4
10 10 maximal 90
extension and flexion
contractions at 180/sec on
Kin-Com
Carnosine in both
trained and untrained
legs with -ALA
Training alone had
no effect on
carnosine levels
None measured.
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88
Table 3. Cont.
Smith et
al., 2009
[59]
46
recreational
ly active
young
males
6g/day for 3
weeks, then
3g/day for 2
nd
3
weeks of -ALA
or placebo
(dextrose)
High intensity interval
training
Not measured Training
increased
EMG
FT
, no
additive effect
with -ALA
Smith et
al., 2009
[58]
46
recreational
ly active
young
males
6g/day for 3
weeks, then
3g/day for 2
nd
3
weeks of -ALA
or placebo
(dextrose)
High intensity interval
training
Not measured VO
2
peak
and time to
reach
VO
2
peak
with -ALA
lean body
mass with -
ALA

3.4. Beta-Alanine and Muscular Fatigue

There are several factors that play a role in muscular fatigue with high-intensity exercise. Some
common theories include a disruption of the neuromuscular junction; a decrease in Ca
2+
release and
uptake leading to the inability of muscles to contract; a depletion of fuel stores such as ATP;
production of free radicals due to oxidative stress; and, the accumulation of metabolites such as
H
+
[38]. Carnosine has been implicated to play a role in each of these proposed mechanisms of fatigue,
but is most commonly researched for its effect on metabolite accumulation as a buffer.
The previously mentioned study by Derave et al. [5] also examined the effects of -ALA
supplementation on isokinetic and isometric fatigue. The isokinetic protocol involved performing five
sets of 30 maximal voluntary isokinetic knee extensions at 180/sec with one minute of recovery
between sets on the right leg. The isometric protocol was performed on the left leg and involved a
maximal static voluntary contraction (MVC) at 45. Once the MVC was determined, subjects
performed isometric contractions at 45% of the MVC for as long as possible. Results indicated that
carnosine loading significantly improved the latter stages of exercise (sets four and five of the
isokinetic test). The researchers noted that the observed response with -ALA supplementation had
similar results as muscle creatine loading on muscle fatigue [66]. The authors also suggested the
increase in carnosine attenuated fatigue by not only its buffering capacities, but also by its ability to
improve myofibrilar Ca
2+
sensitivity.
Neuromuscular fatigue is defined as an increase in electrical activity of a working muscle over
time [67,68,69]. The increase in electrical activity is observed by the increase in EMG amplitude and is
indicative of the recruitment of more motor units and/or the increase in firing rate of the active motor
units in order to attempt and sustain the given activity [69]. The accumulation of H
+
ions is one
possible explanation for this EMG response. Other possible explanations include depleted energy
stores and impaired regulation of muscle cations [2,70]. deVries and coworkers [67] developed a
protocol to assess neuromuscular fatigue threshold. It was termed the PWC
FT
and examines the
relationship between EMG amplitude and fatigue during cycle ergometry. This specifically measures
Nutrients 2010, 2


89
the power output at the point of neuromuscular fatigue [8]. Subsequent studies have shown
relationships between PWC
FT
and VT as well [69,71].
Since it has been established in previous research that -ALA supplementation has enhanced
buffering capabilities during exercise by the subsequent increase in muscle carnosine
content [5,6,7,9,31], it has been hypothesized that -ALA supplementation may delay fatigue [8]. Until
recently, this had only been shown in trained and untrained men [7]. Stout and coworkers [8] examined
the effects of 28 days of -ALA supplementation in women on PWC
FT
, VT, VO
2
max, and TTE during
a cycle ergometry protocol. Subjects were assigned to supplement with either -ALA or placebo
(maltodextrin) in doses of 3.2 g daily for days one through seven and 6.4 g daily for days eight through
28. Subjects were tested prior to and following supplementation. Results showed that -ALA
supplementation increased PWC
FT
by 12.6%, VT by 13.9%, and time to exhaustion by 2.5%.
Stout and colleagues [61] also recently examined the effects of three months of -ALA
supplementation on PWC
FT
in elderly men and women. Participants supplemented with either 2.4 g -
ALA or placebo (microcrystalline cellulose) three times per day for the duration of the study. Results
revealed that -ALA supplementation increased physical working capacity in an elderly population by
28.5%. The researchers attributed these findings to an increase in muscle carnosine concentrations
leading to an enhanced buffering capacity, although carnosine was not directly measured in this
study [61]. The data related to by -ALA and muscular fatigue show promise for improvements with
supplementation, but still requires future research.

3.5. Beta-Alanine and Creatine Supplementation

Approximately 95% of the total creatine found in the body is located in skeletal muscles, of which
40% is free creatine and 60% is phosphorylated creatine [72]. Creatine has several roles in the body
during exercise, with one of the most important being as an energy source for high-intensity exercise
bouts. Performances that require immediate energy (such as maximal sprints) utilize high energy
phosphate, ATP and PCr that are stored in the muscles. The reversible reaction in which this energy is
released is: PCr + ATP creatine kinase ATP + creatine [73]. Creatine supplementation enhances
the initial stores and availability of PCr and therefore, theoretically would enhance the mechanisms of
the phosphagen system used in high-intensity exercise and improve the shuttling of high-energy
phosphates in the creatine phosphate shuttle that may potentially improve anaerobic and aerobic
capacity [74,75].
During short-duration high-intensity exercise, ATP is rapidly consumed to provide energy for the
given activity. In order to continue at the same intensity, the body must quickly resynthesize ATP from
its byproducts. At maximal intensities, this is primarily achieved by anaerobic degradation of PCr and
glycogen. The main function of PCr breakdown in this case is to act as an initial buffer and delay the
reliance on glycogenolysis [66]. The decrease in maximal force production has been linked to PCr
stores in a direct relationship [76]. Creatine supplementation in doses of 2030 g/day have shown to
increase skeletal creatine content by about 20%, where 2030% of this is as PCr [77]. Creatine
supplementation also shows to speed the PCr resynthesis within the first minute of recovery from
intense muscular activity [78].
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90
Creatine supplementation has been extensively studied and is known to have ergogenic properties in
power and strength athletes, with recent studies showing supplementation resulting in increases in
muscular strength, anaerobic power, and body mass [66,79-81]. In fact, the majority of long term
training studies with creatine suggests an ergogenic effect with supplementation in a variety of
populations including trained adolescents, adults and the elderly [11]. For example, Kreider and
colleagues [82] examined the effects of 28 days of creatine supplementation during training for college
football players. Subjects supplemented their diet with either a carbohydrate electrolyte placebo or this
same supplement containing 15.75 g/day creatine monohydrate for 28 days while engaged in
resistance-training and agility exercises. The researchers reported that the group supplementing with
creatine had greater gains in fat free mass, bench press lifting volume, and repetitive sprint
performance on a cycle ergometer compared to the placebo [82]. More recently, studies have examined
the effects of supplementing the diet with creatine monohydrate and -ALA on exercise performance
and training adaptations.
A study by Hoffman and colleagues [12] used male power athletes and supplemented their diet with
creatine or a combination of -ALA and creatine. The supplementation doses were 10.5 g daily of
creatine monohydrate; 10.5 g daily of creatine monohydrate in combination with 3.2 g daily of -ALA;
or, 10.5 g daily of dextrose as a placebo. In addition to supplementation, subjects were involved in a
ten week detailed resistance training program with workouts four days a week. The researchers
reported significant improvements in body composition after ten weeks of the combined
supplementation of -ALA and creatine in conjunction with resistance training compared to creatine
alone or placebo. Additionally, they showed the addition of -ALA to creatine was able to reduce
fatigue rates during training compared to creatine alone. These findings suggest that there may be
additive effects of supplementation of creatine and -ALA.
Stout and coworkers [4] examined the effects of 28 days of -ALA and creatine supplementation on
neuromuscular fatigue and PWC
FT
. In the study, 51 men supplemented their diet with either 34 g of a
dextrose placebo; 5.25 g of creatine with 34 g of dextrose; 1.6 g of -ALA with 34 g of dextrose; or,
1.6 g of -ALA with 5.25 g of creatine and 34 g of dextrose. Subjects ingested this dose four times a
day for the first six days, and then only twice a day for the remainder of the study. Results revealed
that PWC
FT
increased in the -ALA group with no additive effect of creatine. The researchers
suggested that 28 days of -ALA supplementation was able to delay neuromuscular fatigue during
incremental cycling but this was independent of the inclusion of creatine [4].
A study by Zoeller and associates [10] examined the effects of four weeks of creatine and -ALA
supplementation on VO
2
peak, lactate threshold (LT), VT and TTE. This study had four
supplementation groups including a placebo of 34 g dextrose; 5.25 g creatine monohydrate plus 34 g
dextrose; 1.6 g -ALA plus 34 g dextrose; and, a combination of 5.25 g creatine monohydrate and 1.6
g -ALA plus 34 g dextrose. Subjects ingested these supplements four times a day for six days and
then twice a day for the duration of the study. The combined creatine and -ALA supplementation
resulted in significant increases in five of the eight cardiorespiratory endurance variables tested (VO
2

and power output at LT and VT, and percent VO
2
peak at VT). Individually, results revealed
improvements in power output at VT and total TTE for creatine alone group and improvements in
power output at LT for -ALA alone group. However, no significant effects were noted between
groups. Therefore, it was concluded that the combination of creatine and -ALA supplementation may
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91
potentially be beneficial in improving submaximal exercise performance when measured at the lactate
and ventilatory thresholds [10]. Collectively, these findings suggest that there may be benefit of
supplementing the diet with creatine and -ALA but it is unclear whether these benefits are
independent or additive in nature.

3.6. Summary of Beta-Alanine Supplementation

The use of -ALA in recent research has shown to increase muscle carnosine concentrations in as
short as two weeks, with increasing levels with longer supplementation periods [6,55]. However,
although there is strong support that -ALA supplementation during training possesses ergogenic
value, the specific mechanism of action and ergogenic value remains to be fully examined. Some
studies show that -ALA supplementation can improve high intensity exercise capacity, delay VT
and/or neuromuscular fatigue, promoted greater gains in lean body mass during training, and increase
VO
2
peak or time to exhaustion. On the other hand, other studies show limited effects of -ALA
supplementation on exercise performance. The combination of -ALA and creatine monohydrate
supplementation is still a new field of research with conflicting results. Additive effects were shown in
one study for improving fatigue rates with a resistance training program as well as for increasing lean
body mass [12]. Combined supplementation was also shown to improve VT and LT during
incremental cycle exercise [10]. Other studies failed to show additive effects for variables such as
anaerobic power [12] and PWC
FT
[4]. However, dosing patterns differed in these studies so it is
difficult to draw definitive conclusions.

4. Future Directions

Future research is needed to examine the effects of -ALA supplementation on muscle carnosine
concentrations as well as the physiological effects of increasing muscle carnosine. In this regard, more
research should be conducted to understand the effects of -ALA supplementation and corresponding
increases in muscle carnosine concentrations on muscle buffering capacity, antioxidant properties,
enzyme regulation, calcium regulation, exercise capacity, performance outcomes, and neuromuscular
fatigue. An important direction for future research is the determination of an optimal dosing strategy of
-ALA in order to optimize increases in muscle carnosine concentrations, physiological adaptations,
and performance. The current literature shows many variations in the amount and length of -ALA
supplementation; therefore, a standard strategy is still pending. Studies should also examine whether
different types of exercise training may influence muscle carnosine to a greater degree in order to
determine the most effective method of raising carnosine levels. Determining the correct combination
of training and supplementation dose may be especially important in the athletic populations. It will
also be important to study the long-term safety and efficacy of -ALA supplementation.
Further research is clearly warranted to assess the efficacy of -ALA and other ergogenic nutrients
such as creatine. Creatine loading significantly increases muscle phosphagen levels within a few days
whereas it has been determined that -ALA supplementation takes several weeks to increase muscle
carnosine concentrations. Therefore, future research should examine effective dosing strategies to
optimize the benefits of both supplements. It is also possible that different types of athletes may benefit
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92
from both -ALA and creatine supplementation. Therefore, studies need to be conducted to examine
the potential ergogenic value in trained athletes with supplementation. In addition, studies examining
the effects on exercise recovery may be useful since -ALA and creatine supplementation has been
reported to delay fatigue. The majority of current research has focused on the effects in young men,
with the exception of the studies by Stout and associates [8,61] which examined the effects in women
and the elderly. Nevertheless, additional research is needed to examine whether age and/or gender may
influence results. Another area that should be investigated is supplementing the diet with -ALA may
provide some therapeutic benefit for patients with various neuromuscular and/or muscle wasting
diseases as has been reported with creatine supplementation. Finally, additional research should
examine the possible synergistic effects of -ALA with other nutrients.

5. Conclusion

-ALA supplementation is a relatively recent and growing area of research. It carries potential
beneficial effects with high-intensity exercise including anaerobic sprints and resistance training.
There is also potential for additive effects of -ALA and creatine, along with other supplements, to
further enhance the possible ergogenic effects. There are many possibilities for future research
opportunities regarding the use of this supplement. The future of -ALA may potentially open the door
to further improvements in high-intensity exercise and sport performance in a wide range
of individuals.

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