JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL SCIENCES
Emmanuel I. Ugwuja, Nicholas C. Ugwu, Uche E. Nwachi. Hematological and biochemical
parameters of diabetic rats administered either energy drink alone or energy drink mixed
with alcohol. Journal of pharmaceutical and biomedical sciences (J Pharm Biomed Sci.) 2013
October; 35(35): 1808-1813.
The online version of this article, along with updated information and services, is located on
the World Wide Web at: www.jpbms.info
Journal of Pharmaceutical and Biomedical Sciences (J Pharm Biomed Sci.), Member journal.
Committee of Publication ethics (COPE) and Journal donation project (JDP).
ISSN NO- 2230 – 7885
CODEN JPBSCT
NLM Title: J Pharm Biomed Sci.
Emmanuel I. Ugwuja, Nicholas C. Ugwu, Uche E. Nwachi.
Original
article
Hematological and biochemical parameters of diabetic rats administered either
energy drink alone or energy drink mixed with alcohol
Emmanuel I. Ugwuja1,2 *, Nicholas C. Ugwu1, Uche E. Nwachi 3
Affiliation:1 Departments
of Chemical Pathology, Ebonyi State
University, P. M. B. 053 Abakaliki, Nigeria
2 Department of Biochemistry, Faculty of Biological
Sciences, Ebonyi State University, P. M. B. 053
Abakaliki, Nigeria
3 Department of Medical Biochemistry, Faculty of Basic
Medical Sciences, Ebonyi State University, P. M. B. 053
Abakaliki, Nigeria
The name of the Department and Institution to
which the work should be attributed:Departments of Chemical Pathology
Department of Biochemistry
Department of Medical Biochemistry
Ebonyi State University, P. M. B. 053 Abakaliki,
Nigeria
Core Idea:
Consumption of either energy drink alone or in
combination with alcohol is gaining prominence,
especially among the youths and athletes despite
their safety concern. The practice has permeated
the elderly population, who is more vulnerable
to diabetes mellitus, with unverified claims that
they contain low sugar and as well ameliorates the
toxic effect of alcohol. The idea of th e presen t
study was to highlight the haematological and
biochemical alterations associated with such
practices in diabetic model, as no scientific
evidence is available to unsuspecting consumers.
*Correspondence to:
Emmanuel I. Ugwuja.
Departments of Chemical Pathology, Ebonyi
State University, P. M. B. 053 Abakaliki, Nigeria;
E-mail: [email protected]
Abstract:
Consumption of either energy drink alone or in
combination with alcohol is gaini ng popularity
despite concerns of their s afety. The present study
investigated the hematological and biochemical
parameters of diabetic rats administered either
energy drink alone (ED) or energy drink mixed
with alcohol EDmA). Twenty male albino rats
weighi ng 172-185g grouped into four (A-D) of five
rats per group were investi gated. Rats in groups A,
B and C were made diabetic by intraperitoneal
injection of alloxan (200mg/Kg body weight).
Groups A and B rats were administered 3.75ml /Kg
Bullet® and 3.75ml/Kg Bullet® + 1.0g/Kg alcohol,
respectively while rats in groups C and D were
given water only and acted as diabetic (DC) and
non-diabetic (NDC) controls, res pectively. The
experiment lasted for 30 days after which the rats
were killed and their blood collected for
hematological and biochemical parameters using
standard methods. Results showed that di abetic
rats administered either energy drink alone or
energy drink mixed with alcohol exhibi ted
alterations in both haematological and biochemical
parameters, but significant effects were observed
only in rats admi nistered energy drinks mixed with
alcohol. It may be concluded that the biochemical
derangements associated with diabetes mellitus
and/or consumption of energy drink may be
exacerbated by alcohol.
Key words: Energy drinks ; alcohol; diabetes
mellitus, biochemical derangement; dyslipidaemia.
Article citation:Emmanuel I. Ugwuja, Nicholas C. Ugwu,Uche E. Nwachi. Hematological and biochemical
parameters of diabetic rats administered either energy drink alone or energy drink mixed with
alcohol. Journal of pharmaceutical and biomedical sciences (J Pharm Biomed Sci.) 2013 October; 35(35):
1808-1813. Available at http: //www.jpbms.info.
1808
ISSN NO- 2230 – 7885
CODEN JPBSCT
NLM Title: J Pharm Biomed Sci.
Emmanuel I. Ugwuja, Nicholas C. Ugwu, Uche E. Nwachi.
INTRODUCTION
onsumption of energy drinks alone or in
combination with alcohol is becoming a
common practice, especially among the
athl etes, teenagers and students 1. It is estimated
that energy drink cons umption among adol escent
and middle-aged popul ation between 2001-2008
ranged from 24-56% 2.
Also, a survey of energy
drink consumption among student athletes in
Ghana rev ealed that 62.2% consumed at l east one
can of energy dri nk in a week of which 53.6%
reported that they did so to replenish lost energy
after training and competition3.Other reasons
given for consumption of energy drinks included:
to provide energy and fluid to the body, to improv e
performance, and to reduce fati gue. Recently there
is preponderance of young adults and elderly
consuming energy drink mixed wi th alcohol with
unverified claims that they contai n low sugar and
as well ameliorates the toxic effect of alcohol. For
instance, it has been reported that about 20-40%
of young people consume energy drink mixed wit h
alcohol while partying4. Most energy drinks hav e
large amount of caffeine, tauri ne and sugar with
variable quantities of guarana, ginseng, and B complex vitamins as their constituents 2. Although
reports on the safety of energy drink are still
inconclusive, consumption of energy drinks
contai ning high and unregulated amounts of
caffeine has been associated with serious advers e
effects, such as s eizures, diabetes mellitus, cardiac
abnormaliti es or mood and behavioural disorders,
particularly in children, adolescents and young
adults and thos e who take certain medications 5.
With the consumption of either energy drink alone
or in combination with alcohol gaining popularity
among the young and elderly peopl e there is need
to provide scientific evidence of their safety or
otherwise. Previous study has shown that rats
administered energy dri nk had alterations in thei r
biochemical parameters without histopathological
abnormaliti es of the brain, heart and liver6.
We hypothesise that biochemical alterations in
diabetic rats administered either energy dri nk
alone or energy drink mixed with alcohol would be
exacerbated. Hence the present study aimed at
evaluating the haematological and biochemical
parameters of diabetic rats administered either
energy drink alone or energy drink mixed with
alcohol.
C
MATERIALS AND METHODS
Animals and induction of diabetes mellitus:
1809
Twenty male albino rats wei ghing 172-185g used
for this study were purchased from animal hous e
of the Department of Pharmacy, University of
Nigeri a Nsukka. The rats were randomly assigned
into four groups (A-D) of five rats per group in a
cage and housed in the animal hous e of
Biochemistry Department of Ebonyi State
University, Abakaliki. The animals were allowed to
acclimatise for 7 days under standard
environmental conditions and maintained on a
regular livestock feed (Pfizer Plc, Lagos, Nigeri a)
and water ad libitum. Rats in groups A, B and C
were made diabetic by intraperitoneal injection of
200 mg/Kg body weight of alloxan dissolved in
distilled water. Fasting blood glucose levels were
measured after three days of alloxan i njection with
a glucometer (ACCUTREND GC (Boerhinger,
Mannheim, Germany), using blood from the tail
tips and diabetes mellitus was confi rmed by
elevated blood glucose >7.8 mmol/l. Using a
modified protocol of Ferreira et al. , Seven(7) rats
in groups A (ED) were orally administered energy
drink only (3.75ml/Kg Bullet®) while group B rats
(EDmA) were admi nistered energy drink mixed
with alcohol (3.75ml/Kg Bullet® + 1.0g/Kg
alcohol). Rats in groups C (DC) and D (ND C) acted
as diabetic and non-diabetic control, respectively
and were given only water. The experiment las ted
for 30 days after which the animals were fasted for
12 hours before they were s acrificed and thei r
blood collected for haematological and biochemical
analyses. All the rats received human care in
accordance with the National Ins titute of Health
guidelines for the care and use of laboratory
animals 8. Blood for hematological parameters and
lipid profile was collected in EDTA bottl es while
blood for other biochemical parameters was
collected in plai n bottles. Immediately the
haematological parameters were compl eted, the
samples were s pun i n a laboratory centrifuge at
2000g for five minutes and plasma and serum
separated and stored at -4°C prior to analysis. All
the analyses were done wi thin 24 hours of sampl e
collection.
Laboratory an alyses:
Hematological parameters including packed cell
volume (PCV), hemoglobin concentration (HBC)
and total white blood cell counts (TWBC) w ere
done as described by Dacie and Lewis 9.Serum
calcium (Ca) was determi ned by atomic absorption
spectrophotometer while sodium (Na+) and
potassium (K+) were determined using flame
ISSN NO- 2230 – 7885
CODEN JPBSCT
NLM Title: J Pharm Biomed Sci.
Emmanuel I. Ugwuja, Nicholas C. Ugwu, Uche E. Nwachi.
photometers. Serum albumin was determined by
colorimetric bromocresol green methods 10 as
described previously, while uric acid was
determined by uricas e method as described by
Fossati et al. 11 Serum urea was determined by the
method described by Jung et al. 12 and creatinine
estimation was done by method originally
described by Benedict and Behie13 and reev aluated
by Stevens et al. 14.Total cholesterol and tri glyceride
concentrations were determi ned by enzymatic
colorimetric assay as described previously 15,16 and
HDL-cholesterol was determined enzymatically
after precipitation of other lipoprotein as
described by Warnic et al. 17 while LDL-cholesterol
was calculated using Friedewald equation18. Serum
Alanine
transaminase
(ALT),
Aspartate
transminas es (AST), alkaline phosphatase (ALP)
and bilirubin were determined using test kits
(Randox Laboratori es, UK) in accordance with
manufacturer’s instructions.
Data analysis:
The data generated were analyzed with Statistical
Package for Social Sciences (SPSS® for Windows®
ver. 16). Results were expressed as mean ±
standard deviation with differences between
means determined by one way analysis of variance
(one-way ANOVA. Statistical significance was
achieved at p-v alues < 0.
RESULTS
Table 1 shows changes in body weight and blood
glucose of the experimental animals. Diabetic rats
in general gai ned wei ght but the weight gain i n
diabetic rats administered either energy dri nk
alone or energy drink mixed with alcohol are
comparable with that of the diabetic control (4. 0 %
and 4.5% vs. 4. 0%, respectively) while non diabetic control rats showed a hi gher weight gai n
(7.4%). Howev er, blood glucose of diabetic rats
administered ei ther energy drink alone or energy
drink mixed wi th alcohol was higher than the
diabetic control (30.5% and 25.0% vs. 22.2%,
respectively) while the non-diabetic control
showed an increase of blood glucos e concentration
of 2.6% only.
Table 1. Effects of consumption of either energy drink alone or energy drink mixed with alcohol on body weight and plasma
glucose of diabetic rats
Experimental groups
Parameters
ED
EDmA
DC
NC
Body weight (g)
Initial
165.6 ± 2.3
170.2 ± 3.5
173.4 ± 2.8
169.8 ± 4.3
Final
172.2 ± 1.5
177.8 ± 2.1
180.4 ± 2.2
182.4 ± 2.6
Change (%)
+6.6 (4.0)
+7.6 (4.5)
+7 (4.0)
+12.6 (7.4)
Plasma glucose (mmol/l
Initial
8.75 ± 0.15
8.92 ± 0.21
8.82 ± 0.23
4.98 ± 1.10
Final
11.42 ± 1.23
11.15 ± 1.21
10.78 ± 0.72
5.11 ± 0.13
Change (%)
2.67 (30.5)
2.23 (25.0)
1.96 (22.2)
0.13 (2.6)
ED: Energy drink alone; EDmA: Energy drink mixed with alcohol; DC: Diabetic control; NDC: Non-diabetic control. Values are
mean ± standard deviation.
From table 2, lower values were obs erved for haematological parameters in diabetic rats administered
either energy drink alone or energy drink mixed with alcohol in comparison to ei ther the di abetic control
or non-di abetic control, although the differences were not statistically significant.
Table 2. Hematological parameters of diabetic rats administered either energy drink alone or energy drink mixed with alcohol
Experimental groups
Parameters
ED
EDmA
DC
NC
PCV (%)
44.3 ± 2.6
41.3 ± 1.7
45.5 ± 5.3
51.0 ± 3.7
HBC (g/dl)
14.7.3 ± 1.2
13.6 ± 0.5
15.1 ± 1.6
16.9 ± 1.2
TWBC (x109/l)
4.9 ± 0.2
5.1 ± 0.5
5.0 ± 0.5
5.4 ± 1.4
PCV: Packed cell volume; HBC: Hemoglobin concentration; TWBC: Total white blood cell count; ED: Energy drink alone;
EDmA: Energy drink mixed with alcohol; DC: Diabetic control; NDC: Non-diabetic control. Values are mean ± standard
deviation .
Table 3 shows the effects of either energy dri nk
alone or energy drink mixed with alcohol.
Significantly (p < 0.05) hi gher serum albumin and
alanine transaminase (ALT) were observ ed i n
1810
diabetic rats administered energy drink mixed
with alcohol in comparison to either the non diabetic and diabetic controls, respectively.
However,
while
comparative
as partate
ISSN NO- 2230 – 7885
CODEN JPBSCT
NLM Title: J Pharm Biomed Sci.
Emmanuel I. Ugwuja, Nicholas C. Ugwu, Uche E. Nwachi.
transaminase (AST) and alkaline phosphatas e
(ALP) activities were observ ed in di abetic rats
administered energy drink alone and the controls
(non-diabetic and diabetic), significantly higher
activities of the two enzymes were recorded in
diabetic rats administered energy drink mixed
with alcohol. In the same vein, while higher (p <
0.05) serum total bilirubi n was recorded in
diabetic rats administered energy drink mixed
with alcohol, comparable v alues were observ ed i n
diabetic rats fed with energy drink alone and the
controls.
Table 3. Liver function parameters of diabetic rats administered either energy drink alone or energy drink mixed with alcohol.
Experimental groups
ED
EDmA
DC
NC
Parameters
Albumin (g/dl)
3.7 ± 1.3a
5.1 ± 0.4b
4.5 ± 0.2a
3.4 ± 0.5a
ALT (U/L)
11.3 ± 2.2a
15.8 ± 1.5b
12.5 ± 1.3a
9.5 ± 1.3a
AST (U/L)
10.0 ± 2.2a
14.0 ± 1.8b
10.5 ± 1.2a
11.5 ± 2.4a
ALP (U/L)
29.8 ± 3.4a
37.0 ± 9.2b
31.5 ± 2.9a
28.8 ± 2.8a
T. Bil (µmol/l)
12.3 ± 0.8a
17.4 ± 2.7b
14.8 ± 2.3a
12.8 ± 1.2a
ED: Energy drink alone; EDmA: Energy drink mixed with alcohol; DC: Diabetic control; NDC: Non-diabetic control. Values are
mean ± standard deviation. Values with different superscript along the row are statistically (p < 0.05) significant.
While serum sodium was not significantly affected
by consumption of either energy drink alone or its
combination with alcohol, significantly higher
values of serum potassium were observ ed i n
diabetic rats administered energy drink mixed
with alcohol and diabetic control in comparison to
either diabetic rats fed energy drink alone or the
non-diabetic control (Table 4). Again, while
comparable serum creatinine was observed in all
the groups, and serum urea was not significantly
different in either diabetic rats fed energy dri nk
alone or the two controls (diabetic and non-
diabetic),significantly higher serum urea was
found in diabetic rats administered a combination
of energy drink and alcohol. Serum calcium did not
show any significant difference among the groups,
but higher (p < 0.05) uric acid level was recorded
in diabetic rats administered energy drink mixed
with alcohol in comparison to those fed energy
drink alone. However, significantly higher s erum
uric acid levels were observed in diabetic and non diabetic control when compared with either
diabetic rats on energy drink alone or those on
energy drink mixed with alcohol (Table 4).
Table 4. Plasma electrolytes, and renal function parameters of diabetic rats administered energy drink alone or energy drink
mixed with alcohol
Experimental groups
ED
EDmA
DC
NC
Parameters
+
a
b
b
K (mmol/l)
3.37 ± 0.60
3.78 ± 0.46
3.89 ± 2.79
3.29 ± 0.24a
Na+ (mmol/l)
100.9 ± 9.8
102.4 ± 3.0
97.5 ± 2.8
99.7 ± 1.7
Ca++ (µmol/l)
2.06 ± 0.13
2.10 ± 0.35
2.02 ± 0.93
1.97 ± 0.27
Urea (mmol/l)
2.40 ± 0.59a
4.03 ± 1.05b
2.60 ± 0.52a
2.35 ± 0.58a
Creatinine (µmol/l)
27.9 ± 4.0
30.4 ± 2.3
26.0 ± 4.6
26.2 ± 5.4
Uric acid (µmol/l)
115.5 ± 6.5a
133.3 ± 10.3b
145.8 ± 0.5c
140.5 ± 22.0c
ED: Energy drink alone; EDmA: Energy drink mixed with alcohol; DC: Diabetic control; NDC: Non-diabetic control. Values are
mean ± standard deviation. Values with different superscript along the row are statistically (p < 0.05) significant.
From Table 5, while consumption of either energy
drink alone or its combination with alcohol has no
significant effect on the plasma total cholesterol,
LDL-cholesterol and triglyceride, significantly
lower (p > 0.05) plasma HDL -cholesterol was
found in diabetic rats administered energy dri nk
mixed wi th alcohol in comparison to other groups.
Table 5. Lipid profile of diabetic rats administered energy drink alone or energy drink mixed with alcohol
Experimental groups
ED
EDmA
DC
NC
Parameters (mmol/l)
Total cholesterol
4.6 ± 0.5
4.6 ± 0.2
5.3 ± 1.9
4.2 ± 0.2
HDL-Cholesterol
0.76 ± 0.12a
0.49 ± 0.10b
0.79 ± 0.07a
0.79 ± 0.12a
LDL-Cholesterol
2.3 ± 0.5
2.0 ± 0.1
2.3 ± 0.3
1.4 ± 0.1
Triglyceride
1.8 ± 0.1
2.0 ± 0.2
1.5 ± 0.1
1.4 ± 0.1
ED: Energy drink alone; EDmA: Energy drink mixed with alcohol; DC: Diabetic control; NDC: Non-diabetic control. Values are
mean ± standard deviation. Values with different superscript along the row are statistically (p < 0.05) significant.
1811
ISSN NO- 2230 – 7885
CODEN JPBSCT
NLM Title: J Pharm Biomed Sci.
Emmanuel I. Ugwuja, Nicholas C. Ugwu, Uche E. Nwachi.
DISCUSSION
Based on critical analyses of literatures on the
safety and efficacy of the use of energy drink, the
International Society of Sport Nutri tion (ISSN)
recently recommended amongst others, that
diabetics and individual with pre-existing
cardiovascular, metabolic, hepato-renal and
neurologic disease who are taking medications that
may be affected by hi gh glycaemic load foods,
caffeine, and or other stimulants should avoid us e
of energy drinks unl ess approved by thei r
Physician19. Data from the present study seem to
support this warning as diabetic rats administered
either energy dri nk alone or energy drink mixed
with alcohol exhibited alterations in both
hematological and biochemical parameters, but
significant effects were observed only for rats
administered energy dri nks mixed with alcohol.
The higher plasma glucose levels and lower values
of hematological parameters observed in rats
administered either ED alone or EDmA in
comparison to diabetic and non-diabetic controls
in the pres ent study, though insignificant, sugges t
possible detrimental effects of either dri nks to the
body systems. For the diabetic rats, consumption of
energy drink may have caused the higher values
for plasma glucose levels in comparison to the non diabetic rats due to absence of insulin to handle the
extra glycemic challenge. On the other hand, the
observed lower hematological parameter (PCV)
reflects the negative association between
hyperglycemia and hematocrit levels 20,21 as no
previous association between energy dri nk
consumption and erythropoiesis was encountered.
The significantly elevated levels of liver function
parameters (ALT, AST, ALP and total bilirubi n)
observed in diabetes rats administered energy
drink mixed wi th alcohol in comparison to thos e
administered energy drink alone and the controls
points to the fact that alcohol worsens biochemical
derangements associated wi th consumption of
energy dri nk. Previously, Ebuehi et al. 6 had
demonstrated that consumption of energy dri nk
was associated with decreased ALT and AST. This
is in contrast to the present fi ndings. In that study
6, consumption of energy drink alone was
investigated, unlike the present study, which
involved consumption of energy drink mixed with
alcohol. It can therefore be argued that the
significant elevation of liver function parameters
observed in rats administered energy drink plus
alcohol in the present study may have been due to
the effect of alcohol. Consumption of energy dri nk
with alcohol has been associated with i ncreased
1812
risk of serious i njury, sexual assault, drink -driving
alcohol-related health consequences.4 Alcohol
consumption has been found to precipitate liver
injury through complex mol ecular mechanisms
including genetic predisposition, nutri tional status,
abnormaliti es in energy metabolism, oxidative
stress, and immunological mechanisms amongs t
other factors 22. The pres ent findings may hav e
public health implications for thos e that are
forming the habi ts of combining energy drink with
alcohol with erroneous belief that it ameliorates
the effects of alcohol. The el evated liver enzymes
observed in rats administered energy drink plus
alcohol sugges ts advers e effects of the drink on the
liver. Although assessment of liver histopathology
was not part of the pres ent study, in futu re studies
this may help unrav el the i nvolvement of liver
injury associated with consumption of energy
drink mixed with alcohol. Interestingly, in contras t
to the findings of Ebuehi et al. 6 which
demonstrated decreased plasma albumi n with
energy drink consumption, in the present study
increased serum albumi n was observ ed in rats
administered energy drink plus alcohol. The
possible explanation may be that the alcohol in the
mixture may have exacerbated dehydration by
excessive diuresis with consequent increase in
serum proteins, including albumin. Thus it could be
inferred that consumption of either energy dri nk
alone or energy drink mixed with alcohol could
worsen the dehydration associated with diabetes.
This may also explain the less wei ght gai n
observed in diabetic rats when compared to the
non-diabetic rats obs erved in the present study.
Significantly elevated plasma potassium in rats
administered energy drink plus alcohol observed
in the present s tudy has important health
implication. Potassium is a very important
electrolyte in heart functioning, as ei ther low or
high s erum potassium levels may lead to cardiac
abnormaliti es. It has previously been reported that
long-term exposure to the various components of
energy beverages may result in significant
alterations in the cardiovascular system 23.
Furthermore, elevated serum urea, and potassium
in rats administered energy drinks plus alcohol i n
comparison
to
controls
suggests
renal
involvement. Urea is a product of protei n
metabolism, which is accumulated in the blood
when the kidneys are affected. Although the
mechanism by which energy drink mixed with
alcohol precipi tate or/induce cardiac and renal
impairments is not clear, i t may partly be through
ISSN NO- 2230 – 7885
CODEN JPBSCT
NLM Title: J Pharm Biomed Sci.
Emmanuel I. Ugwuja, Nicholas C. Ugwu, Uche E. Nwachi.
alterations in serum potassium. Hence it may
equally be argued that the cardi ac abnormalities
previously associated with consumption of energy
drink mixed with alcohol might in part have been
mediated through the alteration in s erum
potassium.
Agai n, in the present study, diabetic rats
administered energy drink mixed with alcohol had
significantly
lower
HDL -cholesterol
(good
cholesterol) in comparison to diabetic rats
administered energy drink alone or the controls.
Hence i n addition to perturbation of s erum
potassium, the cardiac abnormalities previously
associated wi th consumption of energy drink plus
alcohol may also in part hav e been due to disorder
in lipid metabolism as dyslipidaemia has been
associated with cardiac abnormalities 24. It may be
speculated that the reduced HDL-cholesterol
observed in the present s tudy is due to the effects
of alcohol as comparable values were observed for
rats administered energy dink alone, the di abetic
control and the non-di abetic control.
CONCLUSION
It may therefore be concluded that alcohol i n a
mixture of energy drink and alcohol may worsen
biochemical derangements associated with
consumption of energy dri nk or/and in diabetics.
We advocate intensive public health education to
correct the already formed misconception of lack
of adv erse effects of either energy dri nks alone or
its combination with alcohol.
REFERENCES
1.Usman A and Jawaid A: Hypertension in a young boy: an
energy drink effect. BMC Res Notes 2012; 5: 591.
2.Ballard SL, Wellborn-Kim JJ and Clauson KA: Effects of
commercial energy drink consumption on athletic performance
and body composition. Phys Sportsmed 2010; 38: 107-17.
3.Buxton C and Hagan JE: A survey of energy drinks
consumption practices among student -athletes in Ghana:
lessons for developing health education intervention
programmes. J Int Soc Sports Nutr.2012; 9: 9.
4.Bigard AX: Risk of energy drinks in youths. Arch Pediatr.
2010; 17: 1625-31.
5.Seifert SM, Schaechter JL, Hershorin ER and Lipshultz SE:
Health effects of energy drinks on children, adolescents, and
young adults. Paediatrics 2011; 127: 511-28.
6.Ebuehi OA, Ajayl OE, Onyeulor AL and Awelimobor D: Effects
of oral administration of energy drinks on blood chemistry,
tissue histology and brain acetylcholine in rabbits. Nig Q J Hosp
Med. 2011; 21: 29-34.
7.Ferreira SE, de Mello MT, Ross MV and Souza-Formigoni MLO:
Does an Energy Drink Modify the Effects of Alcohol in a Maximal
Effort Test? Alcoholism: Clinical and Experimental Research
2004; 28: 1408–1412.
8.National Research Council (NRC): Guide for the care and use
of laboratory animals. Publication No. 8523 (Rev), National
Institute of Health, Bethesda, MD, 1985.
9.Dacie JV and Lewis SM: Practical haematology 8th edition
Edinburgh, Churchill Livingstone, 1995.
10.Hill PG: The measurement of albumin in serum and plasma.
Ann Clin Biochem 1985; 22: 565-578.
11.Fossati P, Prencipe L and Berti G: Use of 3, 5- dichloro-2hydroxybenzenesulfonicacid/4-aminophenazone chromogenic
system in direct enzymic assay of uric acid in serum and urine.
Clin Chem 1980; 26: 227-231.
12.Jung D, Briggs N, Erickson J, Ledyard P: New colorimetric
reaction for end-point continuous flow, and kinetic
measurement of urea. Clin Chem 1975; 21: 11-36.
13.Benedict SR, Behre JA: Some applications of a new colour
reaction for creatinine. J Biol Chem 1936; 114: 515-532.
14.Stevens JF, Tsang W, Neewall, RG: Measurement of bilirubin,
cholesterol and creatinine in serum and plasma by solid phase
reflectance spectroscopy. J Clin Path; 1983; 36: 598-601.
15.Siedel J, Hegele EO, Ziegenhom J and Wahlefeld AW: Reagent
for the enzymatic determination of total serum cholesterol with
improved lypolytic efficiency. Clin Chem 1983; 29: 1075-1080.
16.Negele U, Hagele EO, Sauer G, Wiedemann E, Lehmann P,
Wahlefeld AW and Gruber W: Reagent for enzymatic
determination of serum total triglyceride with improved
lipolytic efficiency J Clin Chem Clin Biochem 1984; 22: 165-174.
17.Warnic GR, Benderson JM and Alberts JJ: Quantitation of
high-density lipoprotein subclass after separation by dextran
sulphate and Mg2+ precipitation. Clin Chem 1982; 28: 1574.
18.Friedwald WT, Levy RI and Fredrickson DS: Estimation of
the concentration of low-density lipoprotein cholesterol in
plasma without use of the preparative ultracentrifuge. Clin
Chem 1972; 18: 499-502.
19.Campbell B, Wilborn C, La Bounty P, Taylor L, Nelson MT,
Greenwood M, Ziegenfuss TN, Lopez HL, Hoffman JR, Stout JR,
Schmitz S, Collins R, Kalman DS, Antonio J and Kreider RB:
International Society of Sports Nutrition position stand: energy
drinks. J Int Soc Sports Nutr. 2013; 10: 1.
20.Dungab K, Chapman J, Braithwaite S and Buse J: Glucose
measurement: Cofounding issues in setting target for inpatient
management. Diabetes Care 2007; 30: 403-409.
21.Sidebottom RA, Williams PR and Kanavek KS:
Glucose
determination in plasma and serum: Potential error related to
increased haematocrit. Clin Chem 1982; 28: 190-192.
22.Gramenzi A, caputo F, Biselli M, Kuria F, Loggi E, andreone P
and Bernardi M: Alcoholic liver disease: pathophysiological
aspects and risk factors. Alimentary Pharmacology and
Therapeutics 2006; 24: 1151-1161.
23.Higgins JP, Tuttle TD and Higgins CL: Energy beverages:
content and safety. Mayo Clin Proc. 2010; 85: 1033-41.
24.Idemudia JO and Ugwuja EI: Plasma lipid profiles in
hypertensive Nigerians. The Internet Journal of Cardiovascular
Research. 2009; 6: 2.
Copyright © 2013 Emmanuel I. Ugwuja, Nicholas C. Ugwu,Uche E. Nwachi. This is an open access article distributed under
the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium,
provided the original work is properly cited.
1813