Technical Report

COMPARISON OF DIFFERENT SOFT DRINKS

AND THEIR EFFECTS ON HEALTH

By

SHAHNILA YASMEEN
B.Sc. (IUB)

2019-ag-3091

Advised by

Muhammad Ismail Hashmi

Master of Science
In

Chemistry
SUB-CAMPUS BUREWALA-VEHARI
UNIVERSITY OF AGRICULTURE
FAISALABAD-PAKISTAN
2021
TITLE: COMPARISON OF DIFFERENT SOFT DRINKS AND
THEIR EFFECTS ON HEALTH

SIGNATURES

Student _________________

(Shahnila Yasmeen)

EVALUATION COMMITTEE

1. Advisor _____________________

(M. Ismail Hashmi)

2. Member _____________________

(Tabinda Tariq)

3. Principal’s Nominee _____________________

ii
 DECELERATION
I hereby declare that the content of my technical report, entitled “Comparison of
different Soft Drinks and their Effects on health” are product of my own research
and no part has been copied from any published source. I further declare that this
work has not been submitted for award of any other diploma/degree. The university
may take action if the information provided is found inaccurate.

Shahnila Yasmeen

2019-ag-3091

iii
 DEDICATION
Blessed by ALMIGHTY ALLAH

This technical report would be incomplete

Without mention of the

Support given to me by my

‘Parents’

To whom this technical report is dedicated.

They are my own

“Soul out of my soul”

Who kept my spirits up when the muses failed me.

iv
 Acknowledgements
In the name of Allah, the most Gracious, the most Merciful, Praise to Allah
Almighty, the Lord of the world, who addressed his Holy Prophet Muhammad
(PBUH) by saying; “Read! In the name of your Lord, who has created (all that
exists). To esteem the highness of Allah Almighty, I feel myself unapt as my words
have lost their expressions, knowledge is lacking and diction is too short to express
gratitude in the rightful manner to the blessing and support of Allah almighty whose
help had flourished my ambitions and helped me to attain goals. Never think that any
request you have is too much for Allah. He says: “BE‘’ and it is.

Very special thanks to worthy Principal Dr. Sajjid Mahmood Nadeem, and
respected HOD Dr. Rashid Waqas for providing us a comfortable environment at
this campus and department throughout the degree.

The work presented in this manuscript is accomplished under the sympathetic

attitude, observant pursuit, cheering perspective and enlightened supervision of
Respected Sir, Ismail Hashmi, Lecturer, UAF SUB-campus Burewala, he gave me
full-fledge time. I have no appropriate words to express my deep gratitude to
honorable Ma’am Tabinda Tariq for her active unfailing support, guidance, a lot of
cooperation, and most Important for her valuable time in spite of her busiest schedule.
This accomplishment might not have been possible without her guidance.

Last but not the least, I owe more thanks to my all class fellows, friends, well-wishers,
lab coworkers, criticizers, grandmothers, her family, all cousins, most caring, loving,
and special personality in my life whom always support me with unconditional love
(from my childhood and still), and especial thanks to my parents, and siblings who
helped and support me during my all studies. I am also privileged to be the daughter
of great Father and Mother who helped me in every walk of life. May Allah give them
a long happy life (AMEEN).

Shahnila Yasmeen

Content Title Page

ACKNOWLEDGEMENTS viii
v
 ABSTRACT ix

1 INTRODUCTION 1

2 LITERATURE REVIEW 7

2.1 NATURAL TREND IN SOFT DRINK CONSUMPTION 9

2.2 TWO REASONS THAT SOFT DRINKS ARE 9

NUTRITIONAL CONCERN
2.3 SUGAR AND SWEETENERS 12

2.4 FLAVORING AND COLORING 13

2.5 MICROBIAL CONTAMINATION 13

2.6 BAD EFFECT OF SOFT DRINKS ON HUMAN HEALTH 14

2.6.1 ASSOCIATION OF SWEETNED BEVERAGE INTAKE 14

WITH HYPERTENTION
2.7 SOFT DRINK CONSUMPTION AND RISK OF 15
SYNDROME
3 MATERIALS AND METHOD 17

31 TEST FOR SUGAR 17

3.1.1 BANDIC TEST 17

3.2 TEST FOR CARBON DIOXIDE 21

3.2.1 LIME WATER PREPARATION 21

3.3 PH MEASUREMENT 23

4 RESULT AND DISCUSSION 26

LITERATURE CITED 30

Table of Content

List of Figures

vi
Table Title Page

2.1 Water Bath 19

2.2 Solution of Copper Sulphate 19

3.3 Bandic Test 20

3.4 Sugar Test for Coca Cola and Pepsi 20

3.5 Sugar Test for Sprite and 7up 21

3.6 Sugar Test for Sting and Dew 21

3.7 Co2 Test for Coca Cola and Pepsi 23

3.8 Co 2 Test for Sprite and 7up 23

3.9 Co2 Test for Dew and Sting 24

3.10 Co2 Test for Big Apple and Fanta 24

3.11 PH of Coca Cola and Pepsi 25

3.12 PH of Sprite and 7up 25

3.13 PH of Dew and Sting 26

3.14 PH of Big Apple and Fanta 26

vii
 List of Tables

Figure Title Page

4.1 Labelling of Samples 28

42 Physicochemical Study 28

4.3 Test for Carbon dioxide 29

4.4 Test for PH 30

viii
 ABSTRACT
Soft drinks are consumed daily in due to its affordability, characteristic taste, and
thirst quenching potential. However, the high demand may compromise the quality of
production with possible contamination of heavy metals which have shown to cause
intoxication and death in humans. This study evaluated some constituents of soft
drinks and investigated the presence of some heavy metal contaminants. The soft
drinks were screened for the presence of sugar, carbon dioxide, phosphate and
alcohols well as the pH and acidity determined. The level of cadmium, mercury and
lead were determined using atomic absorption spectrophotometer. The study
showed the presence of sugar, carbon dioxide, phosphate, and alcohol in the soft
drinks. The soft drinks were acidic in nature, pH ranging from 3 to 5. These results
suggest that soft drinks may be contaminated with heavy metals which constitute a
major public health problem. Thus, quality control is recommended during the
production process especially at the stages of sterilization and purification.
According to PH measurement the PH of Pepsi is very low and badly effect the
human health due to high acidity of stomach. But comparison of different soft drinks
shows sprite has high PH and good for heath. Because it lowers the acidity of
stomach . Carbon dioxide and sugar present in all soft drinks.

x
CHAPTER 1
INTRODUCTION
Soft drinks are defined as water-based flavored drinks usually with added carbon
dioxide and with nutritive, nonnutritive, and/or intense sweeteners with other
permitted food additives”. soft drink is one of the most consumed dinks exist in
various forms and brands and are marketed by different industries across the
country. These drinks are readily consumed on daily bases especially when
3undergoing tedious activities like hard work and sport . Also, with the relatively
affordable prices. They are highly consumed in celebrations such as traditional
marriages, and weddings. The high consumption rate of soft drink is attributed to
the characteristic taste and flavor as well as their thirst quenching potential.
The consumption of non-alcoholic beverages was rated at 159.85 g/person/day in
2007. Soft drink constituents present such as sugar which is responsible for its
sweetness, carbonated water which is water compressed with carbon dioxide to
make it an ultimate thirst quencher and flavorings agents to add flavor to the
drinks . In addition to taste satisfaction, soft drinks contain other constituents such
as vitamins, phosphates, acids, antioxidants, etc. which are of nutritional and health
benefits to the body (Galadima and Garba, 2012).

Modern soft drinks constitute a diverse group of products. They can be classified
in several ways, for example, on the basis of their sugar and fruit juice content,
flavoring, carbonation level, main nonwater ingredients, and functionality. Apart
from drinking water, the most popular types of soft drinks are as follows: (i) ready-
to-drink essence-flavored beverages; (ii) ready-to-drink beverages containing fruits
o or Fruit juice; (iii) beverages ready-to-drink after dilution . Functional drinks are a
rapidly growing subsector of the market and include drinks enriched with juices,
vitamins, and minerals; sports and energy drinks; wellness drinks and
nutraceuticals. Many functional drinks have been developed to provide specific
medical or health benefits, such as promoting heart health, improving immunity and
digestion, and helping to boost energy . The target markets for functional beverages
are diverse, and Products are often tailored towards particular target markets, for
instance, according to age and gender, with a growing focus on children, women,
and seniors (Haussmann et at., 2012).

Soft drinks intended for human consumption are covered by National regulations
based on codes and standards . For example, the Canadian Food and Drug
Regulations prescribe quality, composition, and labelling standards which also apply
to non-alcoholic beverage manufacturers . Japanese legislation includes the Food
Sanitation Act for food additive and labeling of processed foods. Japan also has a
positive list of additives. Australia and New Zealand Standard nonalcoholic beverages
and Soft drinks, defines a number of products and sets certain compositional
requirements for packaged water, electrolyte drinks, brewed soft drinks, and

11
beverages . In the United States, soft drinks are regulated by the food and drug
administration (FDA). Soft drinks ingredients must comply with all applicable FDA
safety requirements (Heckman et al., 2013). Soft drinks typically contain water,
sweetener (8 ÷ 12%, w/v), carbon dioxide (0.3 ÷ 0.6% w/v), acidulants (0.05 ÷ 0.3%
w/v), flavorings (0.1 ÷ 0.5% w/v), colorings (0 ÷ 70 ppm), chemical preservatives
(lawful limits), antioxidants (<100 ppm), and/or foaming agents.

Some types of soft drink use sugar substitutes. However, certain ingredients may be
hazardous to health if consumed in large quantities, and there is widespread concern
generally with regard to preservatives and sweeteners. Therefore, while on the one
hand there is a trend to produce ever wider ranges of more specialist soft drinks, there
is also pressure to minimize the use of, in particular, artificial and synthetic additives
and ingredients (Gruenwald et al., 2011).

Conventional soft drinks contain 90 percent Water, while diet soft drinks may contain
up to 99% water. Drinking water includes trace amounts of various ions which alter
its taste. Soft drinks manufacturers usually use Softened water to prevent off-tastes
from chlorine residues. The most frequent methods of removing water hardness
employ ion-exchange polymers or reverse osmosis. Other approaches include
precipitation methods and sequestration using chelating agents. These procedures
reduce the concentration of metal ions to approximately 50 ppm Mg and Ca. Water
for soft drinks should fulfill the physical, chemical and microbiological criteria for
drinking water (Ristovska et al., 2012).

With the exception of zero calorie products, soft drinks usually contain between 1%
and 12% sugar (w/w). Sucrose, glucose, or fructose, in various forms, is used as
natural carbohydrate sweeteners. The most common natural sweeteners provide
glucose, the primary source of Energy. Sucrose (saccharide) is a disaccharide
composed glucose and fructose molecule. This sugar can preserve and enhance the
flavor of a drink and gives a satisfying sensation (Mortensen et al., 2012).

Overconsumption of sugars can cause negative health effects, such as obesity,

diabetes mellitus, or non-alcoholic fatty liver disease. Natural sweeteners provide 1.5
÷ 4.0 calories per gram and have been linked with weight Gain. Fructose contributes
to the formation of advanced Glycation end products, which may be factors in the
onset of diabetes, hasten aging processes, and cause thickening of artery walls . Since
tagatose is absorbed slowly and only incompletely in the intestine. Due to health,

12
including dental health, concerns, alternative sweeteners are commonly added to soft
drinks which are produced and labelled as containing “no added sugar (Harding et al.,
2016).

Most low-calorie beverages contain intense sweeteners, which have been approved
for use within levels of acceptable daily Intake (ADI) and in accordance with the
appropriate regulations. The most commonly used sweeteners (with maximum
permitted dosage in the EU) are aspartame (600 mg/L), acesulfame K (350 mg/L),
sucralose (300 mg/L) And saccharin (80 mg/L) . Aspartame consists of two amino
acids: L Phenylalanine and L-aspartic acid, esterified to methyl alcohol. This
compound is 200 times sweeter than sucrose and leaves no unpleasant aftertaste.
However, it is unstable at high temperatures. Aspartame is also unstable in aqueous
solutions, where it is gradually converted into diketopiperazine (DKP). In the body,
aspartame is broken down into phenylalanine (about 50% by weight), aspartic acid
(40%), and methanol (10%) (Saulo, 2015).

The carbonation of soft drinks varies from 1.5 to 5 g/L. Carbon dioxide is supplied to
soft drinks manufacturers either in solid form (as dry ice) or in liquid form maintained
under high pressure in heavy steel containers. This process makes the drink more
acidic, which serves to sharpen the flavor and taste . It also helps preserve soft drinks
for longer time . Acidity regulators are used in soft drinks to improve their taste by
balancing the sweetness. Human saliva is almost neutral (pH approximately and
when our taste receptors interact with acids in food or drink this sensation is
perceived as sourness. Acids also play an important role in the natural preservation of
soft drinks . In most of
beverages, citric acid is the first choice for use as an acid regulator, as it has several
additional benefits, such as enhancing the activity of beneficial antioxidants and
adding aroma. Malic acid is used when strong flavor enhancement is required and
mostly in combination with citric acid. Succinic acid has a wide range of
applications, including dairy and desserts, but due to regulations it can only be used in
instant beverages to be prepared at home (Taylor et al.,2017).

Phosphoric acid has a strong effect on pH and Is commonly used to give a specific
taste profile to cola type beverages. Use of phosphoric acid remains controversial,
since it has been associated with adverse health effects. High levels of phosphorus in

13
the blood, referred to As “hypophosphatemia,” can lead to organ damage, such as
kidney. Poor kidney function can raise level of phosphorus in the blood, which in turn
lowers calcium Levels, increasing the risk of brittle bone disease. Moreover, increased
serum phosphorus levels, as well as other mineral abnormalities, can individually and
collectively contribute to vascular calcification and cardiovascular disease . The
EFSA has presented its opinion on the upper intake level of phosphorus, according to
which healthy individuals can tolerate up to 3000 mg/day without negative effects.
However, in some individuals mild gastrointestinal symptoms have been reported
with supplemental intakes of >750 mg phosphorus per day. Phosphoric acid and its
derivatives were permitted in the European Union prior to 2009 and are therefore
included in the program for revaluation of food additives (Mettler et al., 2014).

The use of colorings in soft drinks serves several important functions: (i) making the
product more aesthetically appealing; (ii) helping to correct for natural variations in
colors or for changes during Processing or storage; (iii) contributing to maintaining
the qualities by which the drink is recognized. There are three basic categories of
colorings: natural colors, artificial colors and caramels. Natural colorings can be
extracted from plants, fruits, and vegetables and comprise two main categories:

(1) Yellow to orange carotenoids, extracted from plants; (ii) Bright red to purple
anthocyanin’s obtained commercially from a range of fruits and vegetables. Natural
colorings are also added to soft drinks for their antioxidant properties due to
increasing consumer preference for natural colors, the trend in both the EU and the
US markets in recent years has been for manufacturers and retailers to reduce the use
of artificial colors in their products. The use of colorings in EU countries must
comply with food Additives Regulation . All permitted colors are listed in Annex II
and Annex III with limitations
Specified on their use. The EFSA‘s expert Scientific Panel which deals with food
additives, the ANS Panel, has begun a reassessment of all permitted food
coloring’s. Flavorings, in contrast to colorings, are used in comparatively small
amounts, so consumer exposure is relatively low. European Union legislation defines
the different types of flavorings as natural, natural-identical, and artificial (Siegmund
et al., 2013).

14
Chemical preservatives are used to improve the microbiological stability of soft
drinks. The types of chemical preservatives that can be used depend on the chemical
and physical properties of both the preservative and the beverage. The pH of the
product, the presence of vitamins, the packaging, and the conditions of storage will
determine what types of preservative, if any, should be used to prevent microbial
growth. Sorbates benzoates and dimethyl dicarbonate (DMDC) (E 242) are permitted
in ready-to-drink beverages in Europe. Sorbates are very effective preservatives
against bacteria, yeasts, and molds. The antimicrobial effectiveness of Sorbates
depends on the physical and chemical properties of the beverage. Sorbates and
benzoates are often used in combination, especially in highly acidic drinks. Sorbic
acid affects yeast growth by inhibiting the uptake of amino acids and the function of
sulfhydryl enzymes, while benzoic acid destroys the internal proton level of microbial
cells. Benzoic acid occurs naturally, notably in cranberries Cinnamon, plums, and
currants and has been used to inhibit microbial growth for many years, including non-
alcoholic beverages. Benzoate salts are particularly well suited for use in carbonated,
non-alcoholic, and juice beverages. They are more stable than benzoic acid, more
soluble in water, and work best at pH levels between 2 (Battey et al., 2015).

The three significant growth predictors for spoilage yeasts are the variables: pH level,
potassium Sorbates, and sodium benzoate. However, benzoates react with ascorbic
acid (vitamin C) and form benzene, especially if they are stored for extended periods
at high temperatures. In the United States, the EPA has classified benzene as a known
human carcinogen for all routes of exposure . Despite the fact that producers have
developed methods to prevent or minimize its occurrence, and the frequency and
levels of benzene formation in soft drinks have not represented a risk to public health
in the past, benzoates are being used more rarely in the industry. This is partly due to
new processing techniques, which have reduced the need to use benzoates in soft
drinks production. However, these preservatives are still necessary to
maintain quality in some beverages. DMDC (Dimethyl dicarbonate)is commonly
used as a preservative in cold sterilized soft drinks. DMDC is very reactive and
rapidly breaks down when added to a substrate, such as a water Based beverage
(Steels et al., 2019).

The principal products are methanol and carbon dioxide; however, the methanol
concentrations after treatment with DMDC (Dimethyldicarbonateare of no

15
toxicological concern. DMDC performs a broad range of antimicrobial actions
Against yeasts, mold fungi, and bacteria. Dimethyl dicarbonate penetrates into the cell
and deactivates enzymes, leading to the destruction of the microorganisms. Various
hydrocolloids, such as guar and locust gum, pectin, and xanthan, are used as
stabilizers and thickeners, especially in diet drinks (to improve mouth-feel) and fruit
juice drinks (to reduce phase separation). Antioxidants, most commonly ascorbic acid,
are used to prevent the deterioration of flavors and colors, especially when drinks are
packaged in oxygen-permeable bottles and cartons. Functional drinks, nutraceuticals,
and wellness drinks contain a variety of unconventional ingredients, which Can
include the so-called “super fruit ”. However, due to the high level of consumption
and demand of soft drinks, quality control within the process of Production
especially during sterilization and purification may be compromised and the
quality of soft drinks may be challenging (Pofahl et al., 2015). As such, soft drinks
have been shown to Contain traces of alcohols as a result of microbial contamination .
Also, the presence of some heavy metals such as cadmium, lead, mercury arsenic,
zinc, etc. in soft drinks which may be due to environmental pollution from surface
and underground water, food and fruits utilized during Production have been
reported . Heavy metals are metals that have shown to be harmful and toxic to the
human body and constitute a major public health concern. These metals have the
potential of causing acute and chronic toxicity by various modes of action in both
children and adults . Some heavy metals act as catalyst in oxidative reactions of
biological macromolecules, therefore their toxic effect may lead to oxidative tissue
damage . Others genotoxicity/carcinogenic potential causing chromosomal
aberrations and mutation as well as cancer. One of the major mechanisms by which
heavy metals exert toxic effect is through impairment of cellular respiration. Some of
the heavy metals of health importance include: cadmium, lead, mercury etc. (Fitch
and Keim, 2012).

Cadmium is a heavy metal whose long term accumulation may lead to cancer since it
is a carcinogenic element. Also, over a long period of intake, cadmium may
accumulate in the kidney and liver because of its long biological half life and may
lead to the kidney cancer (Mortensen et al., 2012).

Lead is known to affect humans and animals of all ages but the effects Of lead are
most serious in young Children . The most common childhood presentation of lead

16
poisoning is central neurotoxicity .Other symptoms of childhood lead toxicity
include; anemia, peripheral motor neuropathy, gastrointestinal complaints such as
anorexia, vomiting, and abdominal pain, and growth delay (Louary et al,. 2017).

Mercury is also another critical health hazard. Its intoxication can occur in infants and
adults and has shown to interfere with numerous cellular processes including protein
and nucleic acid synthesis, oxidative stress, calcium homeostasis, and protein
phosphoryl (Godwill et al., 2015).

Zinc is essential metal and it is important for biological methods. It is essential for
good health. About 33 ppm of zinc is present in adults. Zinc is important component
of many enzymes. These enzyme are essential for many physiological functions like
energy metabolism and protein synthesis. Deficiency of zinc cause dermatitis and
dwarfism Anemia, nausea and electrolytic imbalance can cause by excessive intake
(Chittleborough et al., 2011).

Arsenic is a heavy metal . it has toxic effect in inorganic form. Ingestion of arsenic
may cause gastrointestinal disease such as damage to the nervous system, enlarge the
liver, damage brain, disturbance of blood and circulation, color the skin and loss of
sensation in the limbs. Ling term intake of arsenic can cause black foot disease in
which blood vessels are severely damage . It is carcinogenic. It increase the risk of
bladder lungs kidney and skin cancer (Kannel et al., 2001).

Sodium has different and appealing taste. Sodium is found in table salt. Sodium is
necessary for muscle contraction, maintaining PH balance, nerve transmission,
hydration and muscle contraction. Excess of sodium intake is a major factor for
hypertension and related disease. There commended intake of sodium is 2300mgper
day. People suffering from hypertension should consume less amount of sodium
(Winkelmaye et al., 2016).

Potassium is necessary for maintaining good health. Potassium is essential for many
functions. action of kidney on regulating the fluid balance. Kidney filter the blood. It
also promote the movement of fluid both in and out of the cell . Potassium is needed
for enzyme pathway and for various metabolic activities. The recommended daily
allowance for potassium is 470o mg per day. It may cause various cardiovascular
disease (Grundy et al., 2017).

17
Dental caries is a multifactorial disease that is affected by several factors including
salivary few and composition, exposure to fluoride, consumption of dietary sugars,
and by oral hygiene practices (Gonzoález-Aragón Pineda et al., 2019). Regular (non-
diet)-soft drinks excluding bottled waters contain large amounts of sucrose or high-
fructose corn syrup that have cariogenic potential; a typical 350-ml can of regular
carbonated soft drink contains approximately 10 teaspoons (40 g) of these sugars .
Long-term and frequent consumption of regular-soft drinks with high sugar content
may induce dental
caries. Many studies have shown a positive relationship between caries and intake of
soft (Chi and Scott, 2019). The greatest risk for caries development in children is
associated with the consumption of soft drink between meals rather than with meals.
Unfortunately, dental caries is the most common reason for children aged 5-9 years
to be admitted to hospital in UK when poor oral health is largely preventable (Royal
et al., 2019).

Object:

• The object of my study to find the PH of different soft drinks.

• Determined the presence of carbon dioxide.

• Determined the of sugar.

18
CHAPTER 02

LITERATURE REVIEW

Soft drinks are consumed daily in Nigeria due to its affordability, characteristic taste,
and thirst quenching potential. However, the high demand may compromise the
quality of production with possible contamination of heavy metals which have
shown to cause intoxication and death in humans. This study evaluated some
constituents of twenty-six soft drinks in Nigeria and investigated the presence of
some heavy metal contaminants. The soft drinks were screened for the presence of
sugar, carbon dioxide, phosphate and alcohol as well as the pH and acidity
determined. The level of cadmium, mercury and lead were determined using atomic
absorption spectrophotometer. The study showed the presence of sugar, carbon
dioxide, phosphate, and alcohol in the soft drinks. The soft
drinks were acidic in nature, pH ranging from 3 to 5 with a mean of 3.6 and the
acid concentration was relatively low between 3 and 12 g/L with a mean of 8.1 g/L.
Lead was present in all the samples ranging from 0.17 to 3.39 mg/L with a mean of
0.8, mercury was present in 22 samples ranging from 0.29 to 11.32 mg/L with a
mean of 2.08 mg/L while cadmium was present only in one sample (0.149 mg/L).
When compared to EPA （ European potential authority ） WHO （ World health
organization ） and NIS (Nigeria investigated security standards, the levels of the
heavy metal contaminants were above the tolerated limits f good quality drinking
water in most samples (Asiegbu et al., 2011).

These results suggest that soft drinks in Nigeria may be contaminated with heavy
metals which constitute a major public health problem. Modern drinks now
contain carbon dioxide for carbonation. Carbonated soft drinks accounted for the
largest category of these drinks in 2016, with a market share of 38% in the UK
.Carbon dioxide, a common factor to all carbonates, is added to make drinks fzzy.
Other ingredients include water, sugar (sucrose, glucose, and fructose), intense
sweeteners acid (citric acid, malic acid, and phosphoric Acid), fruit juice,
preservatives, favourings, and colors. Currently, low-and no-calorie drinks make up
45% of the category, with a further 5% being mid-calorie . Some drinks are made

19
with concentrates that require dilution to taste by consumers, such as squashes, and
cordials, accounted for the second largest share (22%) of drink consumption in
2016. There is a dominance of low and no-calorie variants within this category
(87%), providing lower calorie refreshment for adults and children alike fruit juice
is 100% pure juice which is made from the fish of fresh fruit or from whole fruit,
depending on the type used. No sugar, sweeteners, preservatives, favourings or
Colorings’ are added to fruit juice (Caprio et al., 2016).

They contain cells or bits of fruit pulps and vitamin C (ascorbic acid). Fruit
juice accounted for 7% of total soft drink consumption in UK Sport drinks are
another popular drinks especially amongst adolescents and young adults and
they contain water, carbohydrate mainly glucose, maltodextrin as well as fructose,
and electrolytes such as sodium, potassium and chloride . The electrolytes are added
to improve palatability and to help maintain electrolyte balance. Sport drinks aim to
prevent dehydration, and enhance the athletic physical performance before, during
or after sporting activity (Caprio et al., 2016).Soft drinks are often high in sugar
content and acidity each gram of sugar contains 4 calories. In addition, they
supply energy only and are of little nutritional benefits (Bucher and Siegrist, 2015)
.Several studies have shown that soft drink with high sugar and acid Content
consumption can contribute to detrimental general and oral health effects on children
and adolescents including an increasing risk of overweight, obesity, type 2 diabetes,
Dental caries and dental erosion (Chi and Scott, 2019).

Obesity has recently emerged as a major global health problem. The World
Health
Organization (WHO) and Scientific Advisory Committee on Nutrition (SACN)
recommend a diet where a maximum 5% of the energy comes from free sugars. The
SACN (2015) reported that nearly a third of children aged 2-15 years living in the UK
are overweight or obese, and that younger generations are becoming obese at earlier
ages and staying so for longer. In the USA, two out of three adults and one out of
three children are overweight or obese with over 18% of 6-19 year olds are above
the 95th percentiles of body mass index (BMI), for age and gender (Ogden et al.,
2014).A rising consumption of sugar-containing soft drinks has been suggested as
a major contributor to the obesity epidemic. The increase in intake of sugar-
containing soft drink has coincided with rising body weights and energy intakes in
several populations. In the USA, the per capital annual consumption of regular soft
drink increased by 86% between 1970 and 1997 alone. During that period of time,
the prevalence of obesity rose by 112%.Overweight and obesity can have major

20
costs for individuals and their families as well as for the health care system（Flagel
et al., 2000）.

It increases the risk of developing type 2 diabetes and heart disease as well as doubles
the risk of dying prematurely (Pischon et al., 2008).Type 2 diabetes has also
emerged as a global public health concern, parallel to the global trends in the
prevalence of obesity. Along with the increased consumption of soft drinks, there
has been a rapid and large increase in the reported incidence of type 2 diabetes
(Greenwood et al., 2014).Vartanian et al., 2011 , describe high consumption of soft
drinks was related to low consumption of milk, calcium, fruit and dietary fibers
contributing to an overall poorer diet. In addition, in two studies by (Whiting et al.,
2001) and (McGartland et al., 2003), the high intake of carbonated soft drinks
during adolescence was significantly associated with reduced bone mineral density
among Girls aged 12 and 15 years. Calcium is found mainly in dairy products and
is an essential nutrient for the structural integrity of bone and for maintaining
bone density throughout life. whereas, carbonated soft drinks contain mostly empty
calories (Whiting et al., 2001).

Energy drinks are often high in cafeine to enhance the mental and physical
performance, improve alertness, and concentration (Bunting et al., 2013). The amount
of cafeine in most of the energy drinks is usually three times the concentration in
cola drinks. They are available in the market of more than 140 countries and are the
fastest growing soft drink sector not only in the USA and UK but also worldwide
(Seifert et al., 2011).Although moderate symptoms in severe cases (Seifert et al.,
2011) consumption of caffeine can be tolerated by most healthy people, studies
Several artificial sweeteners are used to give diet soft drinks a sweet taste without
sugar. They are called sugar substitutes because they provide the sweetness of sugars
without the added calories, thus reducing the risk for obesity, and dental caries.
However, their safety has been controversial (Whitehouse et al.,2008).The
breakdown product of these sweeteners has controversial health and metabolic
effects. Some research has linked the consumption of artifcial sweeteners with
adverse health conditions including obesity, lymphomas, leukemia’s, cancers of
the bladder, and brain, chronic fatigue syndrome, Parkinson`s disease, Alzheimer`s
disease, multiple sclerosis, autism, and systemic lupus (Shenkin et al., 2003).

The carcinogenic potential of artifcial sweeteners, mainly aspartame and saccharine,

has been investigated. Exposure to these chemicals was associated with an
increased risk of brain tumor sand cancer of the bladder, in both male and female
mice, respectively (Olney et al., 1996) . Another sweetener Saccharine was 9 that

21
its high consumption (>400 mg per day) has been associated with adverse effects on
health including anxiety, restlessness, aggression, headaches, and depression. A
prolonged exposure to high intakes of cafeine, levels greater than 500 - 600
mg a day, can result in chronic toxicity leading to nervousness, nausea,
vomiting, seizures and cardiovascular symptoms in severe cases (Seifert et al.,
2011).

2.1. National trend in soft drink consumption

The prevalence of soft drink consumption among youth ages 6 to 17 years increased
48%, from a prevalence of 37% in 1977/1978 to 56% in 1994/1998. Mean intake of
soft drinks more than doubled, from 5 flow to 12 flow per day. Although the home
environment remained the largest source of children’s soft drink access, an
increasing share was obtained from restaurants and fast-food establishments
(+53%), vending machines (+48%), and other sources (+37%). Away from
home sources of soft drink are an important factor for dietitians to consider
when evaluating the dietary intake and nutritional status of youths. A healthful
diet may be conceptualized as one that provides recommended amounts of essential
nutrients without excessive intake of energy or food components (such as fat)
linked to chronic diseases (Washington et al., 2014).

Compared with an ideal of balance, variety, and moderation, the dietary intake of
US children and adolescents is currently less than optimal in several respects.
Notably, the prevalence of obesity among youth has increased dramatically in recent
Years (Callow et al., 2018). Suggesting that energy intake exceeds needs. At the
same time, intake of several important nutrients, Such as calcium and iron, is
less than recommended (Bareeda et al.,2019). National data indicate that while the
quantity of food Children consume may exceed energy needs, its nutrient content is
less than optimal (4). in particular, the Dietary Guidelines for Americans state that
excess intake of foods high in added sugars is a Concern and draw attention
specifically to the role of soft drinks as a major source of added sugars
(Washington et al., 2014).

National dietary intake surveys indicate that consumption of soft drinks among
children and adolescents is high and has been increasing dramatically in recent
decades (Borrudetal.,2010).Soft drink consumption among children increased
41%between 1989/1991 and 1994/1996, from 198 g per day to 279 g per day
(Morton et al., 2015).

2.2.Two reasons that soft drink are nutrition concern

First, soft drink consumption is associated with higher energy intake, which may
place children at risk for excess weight gain and obesity (Harnack et al .,
2017).The body’s ability to compensate for carbohydrate energy ingested In fluid
form is less precise than for carbohydrate energy ingested in solid form (Mattes et

22
al., 2016) thus increasing the risk for excess total energy intake when energy is
consumed in the form of fluid carbohydrates such as soft drinks. A recent
prospective study among children found that 1 serving of soft drinks per day
increased risk of becoming overweight by 60% during the course of 1 year
(Ludwig et al., 2001). Overweight children are also more likely than normal-
weight children to be heavy soft drink consumers (Troiano et al., 2013).

Soft drinks may affect the dietary quality of youth by displacing milk
consumption (Bowman, 2002). Displacement of Milk by soft drinks can reduce
calcium intake among children because milk is the primary source of calcium in
children’s diets (Fleming et al., 2014). Low calcium intake among children and
adolescents may increase their risk of osteoporosis in later life by contributing to
low peak bone mass (Heaney, 2015). Low calcium intake may also contribute to
Increased current risk of bone fracture (2000). One study of adolescent girls found
that soft drink consumers had a threefold increased risk of reporting a previous
bone fracture compared with girls who did not consume soft drinks. High levels of
soft drink consumption may have other undesirable effects on dietary quality. For
example, compared with no consumers, youth who were high soft drink consumers
were less likely to consume 4 flow fruit Juice per day and had lower mean intakes
of vitamin A, vitamin C, riboflavin, and phosphorus, and lower Calcium
phosphorous ratios (Harnack, 2011). Numerous reasons for the increase in soft drink
consumption among youth have been suggested, including Increased use of youth-
targeted advertising and marketing by soft drink companies, larger single-serving
package Sizes, lower prices, increased national trends toward eating at fast-food
and other restaurants that serve soft drinks as the primary beverage, and the
increasing availability of soft drinks in school settings, from vending Machines or in
the school cafeteria (Shallow et al., 2001).

More information is needed about trends in the availability of soft drinks from
different sources to better understand reasons for the increased soft drink
consumption among youth and to assist with planning effective intervention
strategies. The purpose of this study was to examine national trends in soft drink
consumption among youth. Changes over time in the prevalence of soft drink
consumption, amounts consumed, and sources of soft drink availability were
Examined. This paper expands on our previous report of soft drink consumption
trends among youth (Guthrie and Morton, 2000 ） by Providing data on
consumption trends using the most recent available national survey data from
United States department of Agriculture’s (USDA) Continuing Survey of Food
Intakes by Individuals (CSFII) 1994/1996 and data from the USDA’s 1998
Supplemental Children’s Survey, and by providing information on trends in
sources of Soft drink availability among youth. Data for this study were collected
as part of the USDA food consumption surveys (Washington et al., 2008).

23
The 1977/1978 data are from the Nationwide Food Consumption Survey
1977/1978 (NFCS) ( the 1994/1998 data are from the 1994/1996 Continuing Survey
of Food Intakes by Individuals (CSFII) and the 1998 Supplemental Children’s
survey (ROM et al., 2000).

Gout is the most common inflammatory arthritis in men. The overall burden from
this disease remains substantial and is growing Identifying the risk factors that are
modifiable with available measures is an important first step in the prevention and
management Of this painful condition. The doubling of the prevalence5 and
incidence6 of gout over the past few decades in the United States3 4 coincided
with a substantial increase in the consumption of soft drink sand fructose.7 For
example, soft drink consumption in the US increased by 61% in adults from 1977
to 1997,7 and sugar sweetened soft drinks represent the largest single food
source of calories in the US diet.7 8 Fructose consumption has also increased
dramatically since the introduction commercially produced high fructose corn syrup
in 1967,9 and its yearly per capita use has Increased from 0 kg to 29 kg,10-12
whereas naturally occurring fructose consumption has remained relatively stable.13
Conventional dietary recommendations for gout Have focused on restriction of
purine and alcohol intake but with no restriction of sugar sweetened soft drinks.14
15 Although such soft drinks contain low levels of purine they contain large
amounts of fructose, which is the only carbohydrate known to increase uric acid
levels (Mattes et al., 2016).

In humans, acute oral or intravenous administration of fructose results in a rapid

increase in serum levels of uric acid through accentuated degradation of purine
nucleotides16 and increased purine synthesis. This uprate raising effect was found
to be exaggerated in people with hyperuricaemia18 or a history of gout.17 It is
unknown, however, if this acute effect is sustained on a long term basis and
eventually translates into an increased risk of gout. We prospectively evaluated the
relation between intake of sugar sweetened soft drinks and fructose and the
incidence of gout in a cohort of 46 393 men with no
history of gout. This study identified factors that influence regular soda
consumption among 707 female students, aged 13-18 years, attending North Los
angeles County public high schools. Participants completed a group-administered
Theory of Planned Behavior-based questionnaire. Almost all of the participants,
96.3%, reported that they currently drink soda; 50.1% reported drinking 2 glasses of
soda or more per day during the past year. Students reported drinking regular soda
more than diet soda and reported ranking phosphoric acid-containing soda more
than non-phosphoric acid-containing soda (Mattes et al., 2016).

Attitude, subjective norm and perceived behavioral control had statistically

significant positive associations with intention, and were each significant predictors
of intention to drink regular soda and together explained 64% of its variance. The
Strongest predictor was attitude, followed by perceived behavioral control and

24
subjective norm. Our results suggest that efforts to reduce soda consumption among
female adolescents should include parents and friends. It is also important that soda
should not be excessively available at home or widely accessible to teenagers at
schools. Healthy eating messages for adolescents need to be developed and
incorporated into existing and future campaigns to reinforce the perception that
there are other healthier drinks that quench thirst and that taste good as well (Fitch
and Keim, 2012).

2.3. Sugars and Sweeteners

With the exception of zero calorie products, soft drinks usually contain between 1%
and 12% sugar (w/w). Sucrose, glucose, or fructose, in various forms, is used as
natural carbohydrate sweeteners. The most common natural sweeteners provide
glucose, the primary source of energy. Sucrose (saccharides) is a disaccharide
composed of glucose and fructose molecules bound by an 1,2 linkage. This sugar can
preserve and enhance the flavor of a drink and gives a satisfying sensation. Other
natural carbohydrate sweeteners are as follows: trehalose, isomaltulose
(Palatines),and D-tagatose (Ashurst and Hargitt, 2009). Trehalose is a disaccharide
composed of two glucose molecules bound by an 1,1-linkage. This compound is
characterized by high thermostability and a wide pH stability
range. Its relative sweetness is around 45% that of sucrose. The metabolism of
trehalose is similar to other disaccharides: Ingested trehalose is hydrolyzed into
glucose and absorbed in the small intestine (Mortensen, 2006). Isomaltulose, like
saccharide, is a disaccharide of glucose and fructose, but in contrast to sucrose
is joined by an 1,6 glyosidic bond. Isomaltulose is a tooth-friendly disaccharide
with slow energy release, low glycemic index, and mild sweetness .Tagatose has
a structure similar to that of fructose (Ashurst and Hargitt, 2009).

It is almost as sweet as sucrose and has flavor-enhancing properties. Most

ingested
tagatose is fermented by colon microflora, resulting in the production of short-chain
fatty acids, which are then absorbed almost completely and metabolized. AS is
well known, overconsumption of sugars can cause negative health effects, such as
obesity, diabetes mellitus, or non-alcoholic fatty liver disease. Natural sweeteners
provide 1.5 ÷ 4.0 calories per gram and have been linked with weight gain
(Fitch and Keim, 2012).

Fructose contributes to the formation of advance which have been approved for
use within levels of Acceptable daily Intake (ADI) and in accordance with the
appropriate regulations. The most commonly used sweeteners (with Maximum
permitted dosage in the EU) are aspartame (600 mg/L), acesulfame K (350 mg/L),
sucralose (300 mg/L), and saccharin (80 mg/L). Aspartame consists of two amino
acids: L-phenylalanine and L-aspartic acid, esterified to methyl alcohol. This
compound is 200 times sweeter than sucrose and leaves no unpleasant aftertaste.

25
However, it is unstable at High temperatures and therefore is unsuitable for use in
pasteurized beverages (Hausmann, 2009).

Aspartame is also unstable in aqueous solutions, where it is gradually converted

into
diketopiperazine (DKP). In the body, aspartame is broken down into phenylalanine
(about 50% by weight), aspartic acid (40%), and methanol (10%). Soft drinks with
aspartame must carry a label indicating that the product contains phenylalanine,
which can be harmful to individuals with phenylketonuria, who must strictly limit
the intake of this amino acid aspartame is permitted in more than 100 countries in
the World. Authorities that have approved aspartame include the FDA （ Food
and drug administration ） the Agency Franc ¸ aise de Security Sanitaire des
Aliments (AFSSA), and the Joint FAO/WHO Expert committee on food Additives
(JECFA). In 2013, the European Food Safety authority (EFSA) approved aspartame
for use in food and beverages (Saulo, 2005).

2.4. Flavorings and Colorings

The use of colorings in EU countries must comply with food Additives Regulation .
All permitted colors are listed in Annex II and Annex III with limitations specified
on their use. The EFSA‘s（European food security authority） expert Scientific
Panel which deals with food additives, the ANS Panel, has begun a reassessment of
all permitted food colorings. For example, in 2013 the EFSA (European food
security authority) recommended that new tests be carried out to address the
possible genotoxicity of Allure red AC and other “sulphonated monoazo dyes”:
Amaranth, Ponca 4R,Sunset Yellow FCF, Tartrzine, and Azorubine Carmoisine
(Gruenwald 2014). Another coloring, Ponca 4R , is an allergen, which may elicit
intolerance in people allergic to salicylates (aspirin). It is also a histamine liberator,
which may intensify symptoms of asthma. Ponca 4R is even considered
carcinogenic in some countries, including the USA, Norway, and Finland. It is
currently on the Food and Drug Administration‘s (FDA) list of banned substances in
the USA. The EFSA (European food security authority) has decided to decrease the
ADI （ Acceptable daily intake ） for Ponca 4R from 4 mg/kg To 0.7 mg/kg
bodyweight per day (Golden et al., 2015).

Flavorings, in contrast to colorings, are used in comparatively small amounts,

so consumer exposure is relatively low. European Union legislation defines the
different types of flavorings as natural, natural-identical, and artificial. Based on
work by the EFSA（ European food security authority） in 2012 the
European Commission established a list of flavorings which may be used in the
EU. The EFSA has also developed guidelines which specify the data the
industry should submit in order to facilitate the safety evaluation of new flavorings
(Gyawali and Ibrahim, 2014).

2.5. Microbial contamination

26
Microbial contamination of soft drinks usually originates during the production
process. The raw materials, factory environment, microbiological state of the
equipment and packages, and lack of hygiene are all possible factors （Stratford,
2006 ） . Packaging materials such as cans and bottles can also be sources of
contamination. There are two main methods of producing soft beverages. In the
first, the syrup is diluted with water, after which the product is cooled, carbonated,
and bottled. In the second a precise amount of syrup is measured into each bottle,
Which is then filled with carbonated water. The processes of blending of syrups and
mixing with water, container washing, and container filling are all performed almost
entirely by automatic machinery. Returnable bottles are washed in hot alkaline
solutions for a minimum of five minutes and then rinsed thoroughly. Single
use containers are usually air- or water-rinsed before filling. The preparation of
noncarbonated beverages requires similar processes. However, since they lack the
protection against spoilage provided by carbonation, noncarbonated drinks are
usually pasteurized, either in bulk, by continuous flash pasteurization prior to filling,
or in the bottle. Cold aseptic filling is usually used, especially for sport drinks (Park
and Chen, 2009).

The condition of raw materials and the production environment can directly or
indirectly affect the safety of manufactured soft drinks. The following obligatory
systems have been designed to ensure food safety: Good Manufacturing Practice
(GMP), Good Hygienic Practice (GHP),and Hazard Analysis and Critical Control
Point (HACCP). Soft drinks could introduce additional spoilage species. The Most
important microorganisms with their typical effects on contaminated soft drinks are
presented (Sikora, 2008). However, when technological processes go wrong, the
method of beverage production will have a significant impact on the type of spoilage
microflora. Spoilage is a metabolic process that causes beverages to be undesirable
or unacceptable for human consumption, due to changes in sensory characteristics.
Microbial contamination of raw materials can cause odors, gushing, and other
undesirable defects in the final product. Soft drinks are high in
water activity and often rich in vitamins and minerals, so they present an
attractive
environment for microorganism (Stratford, 2006).

Soft drinks can contain different types of microorganism, but aciduria

microorganisms are the only significant spoilage microflora; although new,
sometimes exotic ingredients used (Back, 2005).

2.6. Bad effect of soft drink on human health

2.6.1. Association of Sweetened Beverage Intake with Incident
Hypertension
sugary sodas and the agent used to sweeten them, high fructose corn syrup (HCFS),
have been implicated in the development of a variety of conditions, including

27
dyslipidemia, insulin resistance, diabetes, non-alcoholic liver disease and gout.1 - 4
Cross-sectional studies have reported positive associations between sugar-sweetened
beverage intake and hypertension prevalence.5,6 However, prospective data are
lacking. Although they do not contain fructose, artificially sweetened beverages
(ASBs) may also be associated with the development of adverse metabolic
consequences such as obesity and chronic kidney disease, through unclear
mechanisms.7,8 We performed a prospective investigation of the association between
intake of SSBs and ASBs and incident hypertension in three large cohort studies: The
Nurses health Study I (NHS I), the Nurses‘ Health Study II (NHSII), and the Health
Professionals‘ Follow Up Study (Fowler et al., 2008).
Higher consumption of SSBs (sugary sweetened beverages was associated with a
small
but significantly increased risk for incident hypertension in the two female cohorts,
and a trend toward an increased risk in the male cohort . Compared with
participants who consumed less than one SSB per month, the fully development of
hypertension among in the two female cohorts, and a trend toward an increased risk
in the male cohort . Compared with participants who consumed less than one SSB
per month, the fully development of hypertension among in the two female cohorts,
and a trend toward an increased risk in the male cohort . Compared with
participants who consumed less than one SSB per month, the fully development of
hypertension among participants who drank one or more SSB per day was 95 % CI
in NHS I, 95 % CI in NHS II, and 1.06 (95 % CI 0.99) in HPFS. In a pooled
analysis of all three cohorts, the adjusted HR （ high rate ） for new-onset
hypertension was 1.01 (95 % CI, 0.99-1.03) for participants who consumed 1-4
sugary drinks monthly, 1.06 (95 % CI, 1.03-1.08) for those drinking 2-6 SSBs
weekly, and 1.13 (95 % CI, 1.09-1.17) for those drinking at least one SSB daily,
compared with those who consumed fewer than one SSB per month (Ascherio et al.,
2012).
Consumption of SBs （ Artificially sweetened beverage ） showed similar
associations with hypertension . In NHS I, consumption of ≥ 1 ASB daily was
associated with a fully adjusted HR of 1.11 (95 % CI 1.08-1.14). NHS II and HPFS
results were comparable. Pooled analyses of all three cohorts yielded an adjusted
HR （ Higher rate ） of 1.04 (95 % CI, 1.01-1.07) for 1-4 diet drinks a month,
1.07 (95 % CI, 1.05-1.09) for 2-6 ASBs weekly, and 1.14 (95 % CI, 1.09-1.18) for
at least one ASB daily. In secondary analyses, adjustment for adherence to a low
calorie diet and for total sugar intake produced no change in results. We then
performed a number of secondary analyses to explore whether there were certain
characteristics of both SSBs and ASBs that were uniquely associated with
hypertension. In general, the direct relation between sweetened beverage intake
(whether sugar- or artificially Sweetened) and hypertension was stronger with
intake of cola-containing as compared with non-cola-containing beverages however,
the difference
in the magnitude of the associations was significant only in the NHS I and HPFS
cohorts (p-interaction < 0.001 and 0.04, respectively (Colditz et al., 2016).

28
The association between sweetened beverage intake and hypertension incidence
was markedly stronger for carbonated as compared with non-carbonated beverages in
all three cohorts interactions were <0.001 in NHS I, 0.03 in NHS II, and 0.009 in
HPFS). There was no Significant difference in hypertension risk and intake of
caffeinated versus non-caffeinated beverages. The fructose content of SSBs contain
fructose, which in animal studies has been shown to cause renal damage, increased
gastrointestinal sodium uptake, and endothelial dysfunction,17 - 19 it was
hypothesized that the HFCS in sugary beverages was responsible for the
increased risk of incident hypertension. In a prospective study, however,
Forman et al. found no association between total fructose intake (regardless of
Source) and hypertension risk.20 Another study measuring inflammatory
markers and reactive oxygen species (ROS) after sugar intake showed no increase
in inflammation or ROS are active oxygen species ingesting orange juice or a
fructose-containing our data indicate that higher intake of both SSBs and ASBs
is independently associated with an increased risk of developing
hypertension, the assumption that fructose in sweetened beverages is central to
their association with elevated blood pressure. It is hypothesize that an ingredient
common to both sugary and diet beverages could be responsible for the increased
risk of new-onset hypertension seen in consumers of soft drinks. Consumption of
carbonated beverages carried a significantly higher risk of incident high blood
pressure than intake of noncarbonated drinks (Nettleton et al., 2009).
2.7. Soft Drink Consumption and Risk of syndrome
The metabolic syndrome was considered present if 3 or more of the following
individual components were present14,22: waist circumference 35 inches (88 cm)
for women or 40 inches (102 cm) for men; fasting blood sugar 100 mg/dL (5.5
mmol/L) or treatment with oral hypoglycemic agents or insulin; blood pressure
135/ 85 mm Hg or treatment for hypertension; serum triglycerides 150 Mg/dL
(1.7 mmol/L) or treatment for hypertriglyceridemia (with Niacin or fibrates);
and HDL-C 40 mg/dL (1.03 mmol/L) in men or 50 mg/dL (1.3 mmol/L) in
women. To assess the relations of soft drink consumption to the incidence of
metabolic syndrome, we excluded participants with prevalent metabolic syndrome
at each of examination cycles 4, 5, and 6(n-2897 person-observations). Then, we
used pooled logistic regression analyses by combining each 4-year follow-up period
of observations to relate the number of soft drinks consumed per day(examination
cola questionnaire) to the incidence of metabolic syndrome. The eligible participants
were free of metabolic syndrome at each baseline examination, and in this setting,
pooled logistic regression has been shown to provide risk estimates similar to time
dependent Cox models (Vereecken et al., 2005).

We compared the consumption of soft drinks 1 per day with infrequent drinkers (1
per day; referent) and also tested for a dose response by comparing groups
consuming 1 and 2 soft drinks per day with the referent group. We Evaluated 2
sets of models which paralleled the analyses of prevalence of metabolic syndrome.
Consumption of soft drinks varies with age and by sex. It has also been suggested

29
that the effects of soft drinks and carbohydrates on metabolic traits may vary
according to age, sex, 26 and baseline body weight. Therefore, we assessed for
effect modification by age (modeled as a continuous variable), sex, and body mass
index (30 versus 30 kg/m2) by incorporating appropriate interaction terms in the
multivariable models. We repeated analyses with additionally adjustment for
alcohol consumption and baseline levels of systolic and diastolic blood pressure,
blood glucose and serum triglycerides. These models were constructed to
account for baseline levels of metabolic traits. Additionally, we repeated analyses
to examine the association between consumption of caffeinated and decaffeinated
soft drinks, considered separately, and incidence of the metabolic syndrome
(Storey et al., 2017).

Because individuals with diabetes mellitus are a particularly high-risk group

for developing metabolic abnormalities, we also repeated our analyses after
excluding those with prevalent diabetes mellitus at baseline. IN our large
community-based sample of middle-aged adults soft drink consumption was
associated with higher risk of developing adverse metabolic traits and the metabolic
syndrome. The present observational data raise the possibility that public health
policy measures to limit the rising consumption of soft drinks in the community
may be associated with a lowering of the burden of metabolic risk factors in adults
(Parks et al., 2010).

Soft drinks play an important role in our life. We use it in different events like
marriage, eid , engagement, daily lunch and in many events. Soft drink is one of the
most consumed beverages. Drinks exist in various forms and brands and are marketed
by different brewery industries across the country . These drinks are readily
consumed on daily bases especially when undergoing tedious activities like hard
work and sport also, with the relatively affordable price. They are highly consumed
during leisure and relaxation. The high consumption rate of soft drink is attributed
to he characteristic taste and flavor as well as their thirst quenching potential but
also responsible of various disease (Cabrera et al., 2019).

30
CHAPTER 3
MATERIALS AND METHODS
The object of this chapter to analysis the different sample of soft drink like coca
cola, pepsi,7up,spirite,,fanta,dew, sting and big apple. The main purpose of this
chapter to analysis the presence of co2, presence of sugar and also determined the
PH of different sample .

3.1 Test for sugar

Test of sugar is done by Bandic test. In order to prepare Bandic solution different
apparatus is used.

Apparatus
▪ 16 beaker of 50 ml
▪ Stirrer
▪ Water bath
▪ Measuring cylinder

Chemicals
 Sodium citrate
 Sodium carbonate

 Distilled water

31
3.1.2 Procedure for Bandic test
Solution A preparation
1. I took 8 beaker of 50 ml , wash with water and dry with the help of
tissue paper. I Weight the 3.46 g of sodium citrate eight time with the
help of weight machine. I added the 3.46 g of sodium citrate in each
beaker.
2. I weight the 2g of sodium carbonate eight time with weight machine
and add into each beaker.
3. I took 8 measuring cylinder and add distilled water to the point of
20ml then I add 15 ml of distilled water into each beaker and place on the
water bath.
Principle of water bath
The principle of water bath is “ when we avoid the direct heating of our
sample on Burner. “Because the direct heating of acidic solution may be
explosive and dangerous.
Water bath firstly on and filled with distilled water and temperature set 60
calcius degree for 30 minute.

Figure 3.1 Water bath

4. After 30 minute ,I remove the beaker from the water bath and place for cooling.
solution A is prepared.
Solution B Preparation
1. In order to prepare B solution I again took the 8 beaker wash with water and
dry with tissue paper.
2. I weight the 0.346 g of copper sulphate eight time and add into each beaker.
. Then I add 5 ml of remaining distilled water in measuring cylinder into copper sulphate.

32
Figure 3.2 Solution of copper sulphate

I add the solution of copper sulphate into the solution A slowly with continuous
stirrer until blue color appear. I add the solution of copper sulphate into the solution
A slowly with continuous stirrer until blue color appear.

Figure 3.3 Bandic Solution

Test of coca cola and Pepsi for sugar

（1）I took two beakers ,wash with water and dry with tissue paper.

（2）I took 3 ml of coca cola and Pepsi sample into each beaker.

（3）I add 3ml coca cola and Pepsi sample into prepared Bandic solution.

（ 4 ） I place this beaker on water bath for 30 minute at 60 calcius degree

temperature.

（5）After 30 minute , I remove the beakers from the water bath.

33
Figure 3.4 Sugar test for coca cola and Pepsi

Test of 7up and sprite for sugar

（1） I took 2 beakers, wash with water and dry with tissue paper.

（2） I took 3ml of each sample into beakers.

（3） I added my sample into prepared Bandic solution.

（4） I place my beakers on water bath for 30 minute on temperature of 60

calcius degree.

Figure 3.5 Test for sugar in sprite and 7up

Test of sting and dew for sugar

（1） I took 2 beakers, wash with water and dry with tissue paper.

（2） I took 3ml of sting and dew sample in each beaker.

（3） I added my samples in prepared Bandic solution.

（4） I place my beakers in water bath for 30 minute at temperature of 60

calcius degree.

（5） After I removed my beakers.

34
 Figure 3.6 Sugar test for sting and dew

Test of big apple for sugar

(1) I took one beaker wash with water and dry with tissue paper.

(2) I took 3ml of big apple sample in beaker.

(3) I added my sample into prepared solution of Bandic solution.

(4) I place my beaker on water bath for 30 minutes at a temperature 60 calcius

degree.

(5) After 30 minutes I removed my beaker.

3.2 Test for carbon dioxide

Test of carbon dioxide done by lime water preparation. So I need different apparatus

Apparatus

▪ 16 Test tubes

▪ Stirrer

▪ Test tube stand

▪ Filter paper

▪ Spatula

▪ Funnel

Chemicals

▪ Distilled water

35
▪ Calcium hydroxide

▪ Sodium carbonate

3.2.1 Lime water preparation

(1) I took 8 test tubes and fill each test tube with half of distilled water and add 2
g of calcium hydroxide.

(2) I place test tubes on test tubes stand.

(3) I took filter paper and adjust on each test tube. I filter my

solution.

(4) Lime water is prepared.

Test of Pepsi and coca cola for co2

(1) I took my samples in test tube and add sodium carbonate in it.

(2) Lime water is added in it.

(3 ) The color turns to milky.

Figure 3.7 Test of Pepsi and coca cola for CO2

Test of sprite and 7up for CO2

(1) I took my samples in test tubes and add sodium carbonate in it.

(2) Then I add prepared lime water in it.

(3) The turns to milky.

36
 Figure 3.8Test of sprite and 7up for CO2

Test of Dew and Sting for CO2

(1) I took my samples in test tubes and add sodium carbonate in it.

(2) I added my prepared limewater in it

(3) It turns slightly change.

Figure 3.9.Test of dew and sting for CO2

Test of big apple and Fanta

（1）I took my samples in test tubes and added sodium carbonate in it.

（2）Then I added my prepared lime water in it.

（3）I turns slightly milky.

Figure 3.10. test of big apple and Fanta for CO

37
3.3 Determination of PH

Apparatus

▪ PH meter

▪ Eight beakers

Chemicals

▪ Distilled water

PH determination of coca cola and Pepsi

（1）I took 3ml my samples in two beakers.

（2） I took PH meter and wash with distilled water.

（3） I dip pH meter into beakers.

Figure 3.11 pH of coca cola and Pepsi

pH of sprite and 7up

(1) I took 3ml of each sample in beakers

(2) I took PH meter and wash with distilled water.

(3) I dip PH meter into each solution.

38
 Figure 3.12. PH of Sprite and 7up

PH of dew and sting

（1）I took 3ml my samples in two beakers.
（2） I took PH meter and wash with distilled water.
（3） I dip pH meter into beakers.

Figure 3.13.PH of Dew and Sting

PH of big and Fanta apple

（1）I took 3ml of each sample in beakers.

（2）I took PH meter and wash with distilled water.

（3）I dip PH meter into solutions

39
Figure.3.14. PH of big apple and Fanta

40
CHAPTER 4
RESULTS AND DISCUSSION
Table4.1 shows the brand names and our given labels to the soft drinks as
SD1,SD2,SD3, SD4, SD5, SD6, SD7 and SD8 ） .Table 4.2 shows the
sugar and reducing sugar content present in soft drinks. The most
common reason for soft drink consumption is the sweetness. Soft drink
show the presence of reducing sugar and non reducing sugar .Both
reducing and non reducing sugar are responsible for its sweetness.
sucrose is absent in some samples confirmed by negative result of Fehling
test.

Table 4.1 Labeling of samples

Sr No. Soft drink brands Abbreviation

1 Coca cola SD1

2 Pepsi SD2

3 7Up SD3
4 Sprite SD4

5 Fanta SD5
6 Dew SD6
7 Sting SD7
8 Big Apple SD8

Physicochemical study table 4.2

Abbreviations Bandic test
1 SD1 +++
2 SD2 +++
3 SD3 +++
4 SD4 +++
5 SD5 +++
6 SD6 +++
7 SD7 +++
8 SD8 +++
（+++ mean abundantly sugar present）

41
Table 4.3 Test for carbon dioxide

Sr No Abbreviation Test for CO2

1 SD 1 +++

2 SD 2 +++

3 SD 3 +++

4 SD 4 +++

5 SD 5 ++

6 SD 6 +++

7 SD 7 ++

8 SD 8 ++

（+++ mean carbon dioxide abundantly ,++ mean moderately mean and +
in lower quantity present）.

42
Table 4.4 PH measurement

Soft drink PH measurement

Sr No.

1 SD 1 2.91

2 SD 2 2.92

3 SD 3 3.35

4 SD 4 3.43

5 SD 5 3.05

6 SD 6 3.14

7 SD 7 3.31

8 SD 8 3.20

43
DISCUSSION
I do many experiments like sugar test, carbon dioxide test and measure
PH by using PH meter there are the following results obtained from
the tables.

▪ Sugar is present in all soft drinks.

▪ Carbon dioxide moderately present in Fanta ,Sting and Big Apple.
▪ Carbon dioxide abundantly present in Coca cola, Pepsi, 7up,
Sprite and Dew.
▪ Coca cola has very low PH as compared to other soft drinks.
▪ Sprite has high PH as compared to other PH.
▪ Sprite is good for health as compared to other soft drinks.
▪ Someone feel fizzy and sooty when drink sprite.
▪ Sprite drink provide hydrate and low the acidity of stomach.
▪ All soft drinks are acidic in nature.
▪Acidic nature is confirmed by PH
measurement.
▪ All soft drinks has PH lower then 7 .
▪ Soft drinks constituents are water ,carbon dioxide and
flavoring agent.
▪ When water and Carbon dioxide react to each other form
carbonic acid.
▪ Carbonic acid in turn give the hydrogen ion.
▪ Soft drinks replace the consumption of milk, which lower the calcium quantity.
▪ Soft drinks overconsumption destroy the stomach wall and give bad
effects on our health.

44
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