Annals of RSCB Vol.

XVIII, Issue 2/2013

RED BULL INDUCES BIOCHEMICAL CHANGES IN WISTAR RAT

LIVER

Mihaela Crişan, Camelia Munteanu, Corina Roşioru, Camelia Lang

FACULTY OF BIOLOGY AND GEOLOGY, BABEŞ-BOLYAI UNIVERSITY,

CLUJ-NAPOCA, ROMANIA

Summary
Despite the increasing popularity of energy drinks’ consumption, there are concerns
regarding the safety and effectiveness of this type of drink. The aim of the present study
was to evaluate the changes in blood and liver biochemical parameters in male Wistar rats,
after 15 days of ad libitum Red Bull energy drink administration. Increased serum
transaminases and LDH activities, and a high glycaemia were observed. An alarmingly
high liver glycogen deposition was also recorded. These results showed that Red Bull
consumption may lead to hepatic injuries and to unexpected metabolic modifications.
Further studies are required to elucidate whether energy drinks are a safe choice for athletic
and cognitive performance enhancement.
Key words: Red Bull, energy drink, liver
camelia.lang@hasdeu.ubbcluj.ro

et al., 2000), but also studies that showed

Introduction
The raising interest in improvement
of sport performance led to an increasing
demand for ergogenic supplements. Energy
drinks, promoted as cognitive and athletic
performance enhancers, are some of the
most popular supplements worldwide.
Red Bull is the best sold energy drink in the
world, with 4.5 billion cans sold per year,
and the number is ever increasing
(http://www.reportbuyer.com).
According to a recent EFSA
(European Food Safety Authority) report,
12% of european adults (18-65 years) and
12% of teenagers (10-18 years) are high
chronic energy drinks consumers, with 7
liters of energy drinks consumed in one
month (aproximately a can of 250 ml/day).
Despite the popularity of these
drinks, their effects on consumers’ health
are quite controversial and not sufficient
research on energy drinks’ safety has been
conducted yet.
There are several studies recording a
modest improvement of energy drinks
consumption on physical endurance (Alford
et al., 2001, Baumm and Weiss, 2001, Seidl
118
no significant enhancement of endurance
related to the consecutive energy drinks
consumption (Carvajal-Sancho and
Moncada-Jiménez, 2005; Eckerson et al.,
2013, etc).
Some of the compounds found in
energy drinks are used in therapy for
treating certain disorders, like taurine and
niacin for dyslipidemias. Other compounds,
like glucuronolactone, are not well studied.
An alarming number of side effects
and even deaths were reported, as a
consequence of energy drinks consumption.
Arrythmia, cardiac arrest and hepatitis are
some of the quoted side effects (Ballard et
al., 2010; Clauson et al., 2008;
Vivekanandarajah et al., 2011).
The present study sought to observe
changes of some biochemical parameters in
liver and serum in Wistar rats, following
Red Bull ingestion ad libitum for 15 days.
This study was designed as the pilot phase
of a more complex project concerning the
effects of Red Bull consumption on
biochemical parameters in liver, brain,
skeletal and cardiac muscle.
Annals of RSCB
Material and methods
All chemicals used in this study
were of analytical grade, purchased from
Sigma-Aldrich Chemie GmbH, Germany,
Nordic Invest S.R.L., Romania and S.C.
BioZyme S.R.L, Romania.
Red Bull energy drink was
purchased from the local market.
16 male Wistar albino rats,
weighing 186.60±3.15 g were used.
Animals were housed in the zoobase of
Molecular Biology & Biotechnology
Department, School of Biology and
Geology, Babeş-Bolyai University, Cluj-
Napoca. They were kept in hygienic
conditions and received a standard diet.
Animals were organized in two
groups: the control group (n=6), received
tap water and the Red Bull group (n=10),
received Red Bull energy drink ad libitum.
After 15 days, the animals were
killed and blood and liver samples were
harvested for biochemical assays:
glycaemia, glucose and glycogen hepatic
content, total serum and liver proteins, ALT
(alanine aminotransferase), AST (aspartate
aminotransferase), LDH (lactate
dehydrogenase) enzymatic activities in
serum.
Blood and liver glucose was
determined using the Somogy-Nelson
method (Somogy, 1945; Nelson, 1944).
Glycogen content was measured using Lo’s
method (Lo et al., 1970). Total protein
concentration was assayed with Bradford
reagent (Bradford, 1976).
ALT and AST activities were
assessed according to the photocolorimetric
method described by Reitman and Frankel
(1957), and for LDH activity, the method of
Bergmeyer and Bernt (1974) was used.
Results were analyzed using two
tailed t test and considered statistically
significant at p ≤ 0.05.
Results and discussion
As expected, Red Bull
administration led to a significant increase
in glycaemia (Fig. 1). A 250 ml can of Red
119
Vol. XVIII, Issue 2/2013
Bull has 27 g carbohydrates, and other
compounds that may affect carbohydrates
metabolism, such as niacin.
According to Zhou et al. (2010) and
Li et al. (2009), niacin supplementation in
high carbohydrate diets may be one of the
causes of diabetes “epidemic”. Moreover,
niacin can induce insulin resistance
(Chang et al., 2006; Greenbaum et al.,
1996).
A can of Red Bull assures 100% of
niacin RDA (Recommended Daily
Allowance). Even if this quantity is not
usually seen as a risk, it should be
considered that high carbohydrate content
of Red Bull may amplify niacin’s side
effects. A normal diet assures niacin RDA
and there is no need for external
supplementation in healthy subjects.
Glycaemia
300
***
250
mg/dl
200
150
100
50
0
Control RedBull
Fig. 1. Glycaemia in rats treated with Red Bull
energy drink ad libitum (n=10) compared with
C group (n=6). The results are expressed as
mean value ± standard error; significant
difference by t test (*** p ≤ 0.005).
Significant liver glucose rise seen in
this experiment (Fig. 2) is also a
consequence of the high carbohydrate
content in Red Bull energy drink. It was
demonstrated that in conditions such as type
2 diabetes, the depletion of skeletal muscle
glycogen is associated with a rise in liver
carbohydrates (Eldar-Finkelman and
Yarkoni, 2002).
Annals of RSCB Vol. XVIII, Issue 2/2013

Liver glucose In the present study, serum and liver

protein concentrations were not affected by
45
40 ** Red Bull administration (Figs. 4 and 5).
35
mg/g tissue

Serum protein
30
25
20 8
15 7
10 6
5

mg/dl
5
0 4
3
Control RedBull 2
1
Fig. 2. Liver glucose in rats treated with Red 0
Bull (** p ≤ 0.01). Control RedBull

The glycogen content in liver was Fig. 4. Serum protein in rats treated with Red
Bull
tripled by Red Bull ad libitum
Liver protein
administration (Fig. 3).
Abnormal glycogen deposition in 450
400
liver due to Red Bull administration has 350
been noted for the first time in the present
mg/g tissue
300
250
study. 200
Hepatic glycogen deposition is seen 150

in 80 % of chronic diabetic patients. It is 100

50
believed that this condition is closely 0

related to liver enzymes abnormalities Control RedBull

(Levinthal and Tavill, 1999).
In a study conducted by Clore et al.
(1992) in humans, after 3 days of fasting,
hepatic glycogen content of type 2 diabetic
patients was 2-fold higher than that of
controls.
Liver glycogen
Fig. 5. Liver protein in rats treated with Red
Bull
As seen below, ingestion of Red
Bull ad libitum in rats led to a significant
rise of serum AST, ALT and LDH
activities (Figs 6, 7, and 8).
AST activity in serum
µ g pyruvate/ ml serum/ hour

250
3 *** *
200
2.5
150
mg/g tissue

1.5 100

1 50
0.5
0
0
Control RedBull
Control RedBull
Fig. 6. Serum AST activity in rats treated with
Fig. 3. Liver glycogen in rats treated with Red with Red Bull energy drink (* p ≤ 0.05).
Bull (*** p ≤ 0.005).

Ebuehi et al. (2011) reported an

increase in serum protein after
administration of 5 ml Red Bull daily, for
36 days, in rabbits.

120
Annals of RSCB Vol. XVIII, Issue 2/2013

ALT activity in serum

350 * References
µ g pyruvate/ g tissue/ hour

300 Akande I.S., Banjoko O.A., Assessment of

250 Biochemical Effect of “Power Horse”
200 Energy Drink on Hepatic, Renal and
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100 Rats, Annu. Rev. & Res. Biol., 1(3): 4556,
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0 Alford C., Cox H., Wescott R., The effects of
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Fig. 7. Serum ALT activity in rats treated with and mood, Amino Acids, 21(2): 139–150,
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Our results are supported by other studies, Carvajal-Sancho A., Moncada-Jiménez J., The
acute effect of an energy drink on physical
which reported hepatic injuries related with
and cognitive performance of male athletes,
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Red Bull sales report,
http://www.reportbuyer.com
EFSA, www.efsa.europa.eu/efsajournal
Red Bull official site, http://www.redbull.com