DEDAN KIMATHI UNIVERSITY OF TECHNOLOGY

ANALYSIS OF CHEMICAL CONSTITUENTS OF SELECTED ENERGY DRINKS

DAN OTIENO ODERO

S080-01-1472/2016

A research proposal submitted to the School of Science in partial fulfillment of the requirements
for the award of the degree of Bachelor of Science in Industrial Chemistry of Dedan Kimathi
University of Technology.
DECLARATION AND RECOMMENDATION

DECLARATION
I declare that this project proposal is my original work and has not been presented in any other
institution for an award of a degree.
DAN OTIENO ODERO
S080-01-1472/2016
Signature………………………. Date…………………………

RECOMMENDATION
This research proposal has been submitted for examination with my approval as the university
supervisor.
Dr. PAUL TANUI
Dedan Kimathi University of Technology
Signature………………………. Date…………………………
DEDICATION
I dedicate this to the Almighty GOD, my parents and those who supported me in one way or the
other.
ACKNOWLEDGEMENT
I would like to thank the Almighty God for giving me energy and knowledge to carry out my
research. Much thanks also goes to the entire Chemistry Department of Dedan Kimathi University
of Technology, for giving me this opportunity to exercise my research project. I would like to
thank Dr. Paul Tanui and Dr. Amdany for their moral support in developing a research topic.
ABSTRACT
This research work examines and compares the physicochemical properties and some chemical
constituents of selected energy drinks. Fourteen (14) brands of energy drinks samples consisting
eleven (11) liquid and three (3) powdered forms will be randomly purchased. All samples will be
analyzed for their physicochemical properties (pH, turbidity, conductivity and total dissolved
solids), trace and heavy metals, aspartame, sugar and caffeine contents. The caffeine and sugar
concentrations will be examined then results recorded.
Contents
DECLARATION ............................................................................................................................ 2
RECOMMENDATION .................................................................................................................. 2
DEDICATION ................................................................................................................................ 3
ACKNOWLEDGEMENT .............................................................................................................. 4
ABSTRACT .................................................................................................................................... 5
LIST OF ACRONYMS .................................................................................................................. 7
CHAPTER ONE ............................................................................................................................. 8
INTRODUCTION ....................................................................................................................... 8
PROBLEM STATEMENT ......................................................................................................... 9
OBJECTIVES ............................................................................................................................. 9
GENERAL OBJECTIVES ...................................................................................................... 9
SPECIFIC OBJECTIVES ........................................................................................................ 9
CHAPTER TWO .......................................................................................................................... 10
REVIEW OF LITERATURE .................................................................................................... 10
CHAPTER THREE ...................................................................................................................... 11
METHODOLOGY .................................................................................................................... 11
PREPARATION OF PHOSPHATE BUFFER FOR MOBILE PHASE .................................. 11
STANDARD SOLUTION PREPARATION ............................................................................ 11
SAMPLE PREPARATION ....................................................................................................... 12
CONCLUSION ............................................................................................................................. 15
REFERENCES ............................................................................................................................. 16
LIST OF ACRONYMS
HPLC High performance liquid chromatography
UV-VIS Ultra violet visible
Ph. Potential Hydrogen
EDs Energy drinks
FDA foods and drinks administration.
CHAPTER ONE

INTRODUCTION
Energy drink products are carbonated, stimulant-laden beverages. They most frequently contain
taurine, arginine, caffeine, acidity regulators, various B vitamins, vitamin C, herbal extracts and
sugar as well as other sweetening substances.
Energy drinks have dose-response relationship with adverse effects; it has become apparent that
the consumption of these energy drinks will seriously harm the body.
Red bull has been known as a healthy drink within many populations. The chemical composition
of energy drinks can produce multiple adverse effects, including serious behavioral effects.
Individuals drink red bull to feel energized during the day. The effects of the drinks are likely
achieved by the amounts of added caffeine and sugar. Simple sugars are metabolized by the body
and produce a quick energy burst, followed by a deep energy deficit. High intake of sugar raises
blood fat levels and leeches’ essential minerals such as copper, chromium and zinc from the body,
leading to deficiency diseases, immune system impairment, and even insulin resistance.
Red bull drinks have many side effects and each 250ml or a can of red bull contains the following;
100mg of taurine, 600mg of glucuronolactone, 80mg of caffeine, 18mg of niacin (niacin amide),
2mg of vitamin B6, 6mg of pantothenic acid (calcium d-pantothenate), Vitamin B2 (riboflavin),
Vitamin B12 inositol, and non-medicinal ingredients: carbonated water, sucrose, glucose (27g of
sugar), citric acid, flavors, and caramel.
Taurine is not essential for humans and it should only be recommended under supervision of a
physician; since glucuronolactone is a precursor to taurine, the body manufactures
glucuronolactone naturally.
Energy drinks increase blood pressure, heart rate or alter glycemic levels; these can often prevent
normal sleep cycles and may have more serious effect on dehydrated athlete by increasing the
heart rate. Consumption by children may induce osseous effects, insomnia, cardiovascular diseases
and mood and behavioral disorders. According to current literature, the risk of negative
consequences is higher certain populations, including children, pregnant women (risk of late
miscarriages, impaired fetal growth) and people with underlying cardiac conditions or diabetes
mellitus.
Caffeine is a most common ingredient of energy drinks. It is added as a flavoring and to make the
drinks addictive. Caffeine is a bitter in taste, white crystalline xanthine alkaloid that’s acts as
psychoactive stimulant drug and a mild diuretic. Almost sixty plant species are known to contain
caffeine. Common sources of caffeine are the (bean) seed of the coffee plant; in the leaves of the
tea bush; and in kola nuts. Caffeine is one of the world’s most widely used drugs. Caffeine is a
naturally occurring substance found in the leaves, seeds or fruits of other 63 plant species
worldwide and it is a part of a group of compounds known as methyl xanthine.
PROBLEM STATEMENT
Energy drinks have been known as health drinks among many people. It has greatly been consumed
by young adults. These are beverages that contain large amounts of caffeine and other stimulants
for example inositol, taurine, glucuronolactone, niacin, panthenol, and many more that are
considered as source of energy. Some of the energy drinks consumed have dose relationship with
adverse effects. It has become apparent that the consumption of these energy drinks will seriously
harm the body. Some of the effects associated with frequent usage of these drinks are renal vascular
congestion, hemorrhage of interstitial tissue, focal atrophy and degeneration of lining epithelium
of proximal and distal convoluted tubules. They can increase blood pressure, heart rate or alter
glycemic levels; these can often prevent normal sleep cycles and may have more serious effect on
dehydrated athlete by increasing the heart rate. There is some concern among health professionals
that these beverages and the drinking behaviors of the targeted consumers may have adverse health
consequences. This is the reason why I came up with this project proposal to evaluate the contents
in these energy drinks and to analyze the amounts of caffeine and other stimulants like sugar used
in the manufacture of these energy drinks to help reduce their consumption thereby reducing the
side effects associated with them thus saving the younger generation.

OBJECTIVES
GENERAL OBJECTIVES
1. To analyze the contents of some selected energy drinks in Kenya.

SPECIFIC OBJECTIVES
1. To analyze the amount of caffeine used in the manufacture of red bull, kabisa, reaktor, and
shark energy drinks in Kenya.
2. To determine carbohydrate( sugar) content in EDs
3. To carry out characterization of isolate using High Performance Liquid Chromatography
(HPLC), pH and quantitative analysis.
CHAPTER TWO

REVIEW OF LITERATURE
In many acute clinical human trials, some over-the counter energy drinks have shown a positive
stimulation of resting energy expenditure. With an increase in energy expenditure at rest, more
energy is being expanded as long as the chemicals are within the system.
A greater amount of calories being expanded at rest translates into weight loss over a period.
The authors concluded from the data that there is a positive effect of a taurine-containing drink on
hormonal responses, which led to a higher endurance performance.
It should be noted that excessive caffeine consumption or caffeine sensitivity can cause a wide
variety of nonthreatening effects such as anxiety, nervousness, irritability, restlessness, headaches
and diarrhea. So it is important to establish the appropriate amount needed.
CHAPTER THREE

METHODOLOGY
Almost ten commercially available energy drinks samples from Nairobi and Nyeri will be
collected for the quantitative determination of their caffeine and sugar content with high
performance liquid chromatography (HPLC). Calibration models to be built with partial
least squares regression (PLSR). An HPLC-UV method is used to measure the reference
values for caffeine contents. The combination of FT-NIR with multidimensional chemo
metric techniques like PLSR can be a good option for the detection of low caffeine
concentrations in energy drinks.
Moreover, three types of energy drinks that contain (i) taurine, (ii) arginine and (iii) none
of these two components will be classified correctly using principal contents analysis and
linear discriminant analysis. Such classifications are important for the detection of
adultered samples and for quality control as well. In this case, more than ten samples will
be used for the evaluation. The classification will be validated with cross validation and
several randomization tests.

Reagents: caffeine standard, sodium carbonate, sodium dodecyl sulfate, Ultra water,
chloroform, tetrahydrofuran (THF), benzoic acid, trimethylamine (TEA).
Real samples; red bull, shark, kabisa, lucozade, polish, coca cola blak, azam, reactor, club
mate.
A total of five samples of energy drinks namely red bull, shark, monster, kabisa, azam will
be purchased from local supermarket in Nyeri.

PREPARATION OF PHOSPHATE BUFFER FOR MOBILE PHASE

50mM of pH 3 phosphate buffer solution is made for the mobile phase by dissolving a
21.305g sample of Na2HPO4 in 1000ml of deionized water and then adding concentrated
HCL drop wise until it reaches pH of 3. The remaining solution was then diluted with
deionized water to final volume of 3L to make a 50mV solution. The solution is then
filtered in order to obtain a phosphate buffer.

STANDARD SOLUTION PREPARATION

A standard solution is prepared by combining 1920ml of the filtered buffer with methanol
to make 60:40 v/v phosphate buffer/methanol solution. A 0.0504g amount of pyridoxine
hydrochloride and 0.1253g amount of caffeine are weighed and transferred to the same
100ml volumetric flask. The flask is then diluted to the mark with the phosphate
buffer/methanol solution and sonicated to dissolve the pyridoxine and caffeine.
SAMPLE PREPARATION
60ml of energy drink is poured into a conical flask and degassed by sonicating for five
minutes. 10ml of the degassed drink is placed in each of the five volumetric flask.
Increasing amounts of standard solution is then placed in each of the five 100ml volumetric
flasks i.e. 1.00ml, 2.00ml, 3.00ml, 4.00ml, 5.00ml. The five volumetric flasks are then
diluted to the mark with the phosphate buffer/methanol solution and mixed thoroughly.
The procedure is then repeated with 3 other energy drinks.
WORK PLAN
 BUDGET

Reagents Price

Caffeine (20g) 500

Pyridoxine hydrochloride (20g) 500

EDs(lucozade, shark, kabisa, reaktor, azam) 1500

Sodium carbonate 750

Sodium hydro phosphate (200g) 300

Methanol 150

Hydrochloric acid 200

Taurine (2-aminoethane sulfonic acid) 350

TOTAL 4250
CONCLUSION
Determination of caffeine content in non-alcoholic beverages and energy drinks is very
important analytical process to safeguard the well-being of people who are unaware to
adverse effects of caffeine.
I will use HILIC/UV/ELSD method for the simultaneous determination of caffeine and
taurine in energy drink. The presented method present the following advantages:
 The method is time efficient as it doesn’t require any sample preparation (beside
sample dilution) and the analysis time is about 8 minutes, resulting in the
possibility of analyzing a large amount of samples in a short time.
 The procedure is simple and inexpensive as only basic LC instrumentation is
necessary.
The developed method is validated in terms of linearity, accuracy and precision. Thus the
proposed method is used to inspect the final product quality as well as for the step by step product
control during the fabrication process.
REFERENCES
1. Almaghamdi, A.H, Almaghamdi, A.F and Alwarthan, A.A (2005). Determination of
contents levels of some food additives in beverages consumed in Riyadh city. J .King
Saud Univ. 18(2): 99-109.
2. Al- Mayaly K.I (2013). Intern. Journal of Research and Dev. In Pharm and life science.
Determination of heavy metals in some artificial fruit juices in Iraqi local markets.
2(4): 507-510
3. Appleton, N “146 reasons why sugar is ruining your health.” Antibiotics to treat.
November 15, 2011
4. Nathanson JA (1984) Caffeine and related methylxanthines: possible naturally
occurring pesticides. Science 226: 184-187
5. Wanyika HN, Gatebe E, Gitau L, Maritim CW (2010) Determination of caffeine
content of tea and instant coffee brands found in Kenyan market. African journal of
Food Science 4: 353-358.
6. Mohammed SG, Al-Hashimi AG, Al-Hussainy KS (2012) Determination of caffeine
and trace minerals contents in soft and energy drinks available in Basrah Markets.
Pakistan journal of nutrition.
7. Hossain M, Jahan I, Akhter S, Raham SMB (2015) Isolation and identification of
Azospirillum isolates from different paddy fields of North Bengal. Indian journal of
Research in Pharmacy and Biotechnology 3:2320-3471.