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Bottled Drinking Water: Assessment of Physical-Chemical and

Microbiological Parameters and Biological Stability of 19 Different

Brands Available in Saudi Arabia

Thesis by

Yasmeen Mohammad Nadreen

In Partial Fulfillment of the Requirements For the Degree of

Master of Science

King Abdullah University of Science and Technology Thuwal, Kingdom of Saudi Arabia

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EXAMINATION COMMITTEE PAGE

The thesis of Yasmeen Mohammad Nadreen is approved by the examination committee.

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COPYRIGHT PAGE

©July, 2021

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ABSTRACT

Bottled Drinking Water: Assessment of Physical-Chemical and

Microbiological Parameters and Biological Stability of 19 Different Brands

Available in Saudi Arabia

Yasmeen Mohammad Nadreen

Bottled drinking water is a common form of water consumption that has grown in popularity and dependency. With countless types and brands available, there are factors to consider regarding the variations in mineral content and microbiological quality, and environmental consequences associated with importing natural bottled waters. Saudi Arabia is the largest desalinated water producer, and although there are scarce natural water resources, consuming locally produced water can alleviate environmental

pressures, so long as local bottled water is of good quality and provides the basic function of drinking water. The objective of this study is to scrutinize the variations in bottled waters available in the Saudi market and compare local and imported waters regarding water quality and compliance with health regulations.

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sparkling, which varied in microbial growth responses. DNA extractions and microbial community analyses were performed on select mineral waters before and after

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ACKNOWLEDGEMENTS

First and foremost, I must thank God for all of the blessings He has bestowed upon me. I am very grateful to Him for this amazing journey and everything that I have learned and experienced.

I would then like to thank Professor Johannes Vrouwenvelder for his great support as my advisor and continuous guidance during my academic journey. I would also like to sincerely thank Dr. Graciela Gonzalez for all of her help, counsel, and insight throughout the course of this research, I appreciate you greatly. Thank you to Professor Pascal Saikaly for your valuable time and feedback.

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TABLE OF CONTENTS

EXAMINATION COMMITTEE PAGE ...2

COPYRIGHT PAGE...3 ABSTRACT ...4 ACKNOWLEDGEMENTS ...6 TABLE OF CONTENTS ...7 LIST OF ABBREVIATIONS ... 10 LIST OF ILLUSTRATIONS ... 11 LIST OF TABLES ... 13 Chapter 1 : Introduction ... 14

Chapter 2 : Literature Review ... 20

2.1 The bottled drinking water industry ... 20

2.1.1 Classifications of bottled water ... 22

2.1.2 Drinking water in Saudi Arabia ... 24

2.2 Environmental impact of bottled water ... 27

2.3 Regulatory Requirements ... 30

2.4 Mineral composition of bottled water ... 31

2.5 Microbiological quality and stability of bottled water ... 33

2.5.1 Regulations for microbiological composition of bottled water ... 33

2.5.2 Microbiological research studies on bottled drinking water ... 34

2.5.3 Determining microbiological content through Flow Cytometry ... 36

Chapter 3 : Surveying Available Bottled Drinking Water ... 39

3.1 Overview ... 39

3.2 Methods ... 39

3.3 Results and Discussion ... 40

3.3.1 Initial bottled water survey findings ... 40

3.3.2 Origin locations of different bottled drinking waters ... 45

3.3.3 Bottled drinking water shelf life ... 48

3.3.4 Price variations among different bottled water samples ... 53

3.3.5 Study implications ... 54

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4.1 Overview ... 56

4.2 Methods ... 56

4.3 Results and Discussion. ... 62

4.3.1 Composition and concentrations of elements listed on bottled water labels ... 62

4.3.2 Analyzing pH values of bottled drinking water samples ... 66

4.3.3 Examining ionic composition of bottled drinking water samples through IC ... 68

4.3.4 Examining ionic and metal composition of bottled drinking water samples through ICP-OES ... 72

4.3.5 Distribution of bottled drinking water types based on mineral compositions ... 80

Chapter 5 : Microbial Content Analysis of Bottled Water ... 82

5.1 Overview ... 82

5.2 Methods ... 83

5.3 Results and Discussion ... 84

5.3.1 Flow Cytometry results for purified bottled water samples ... 86

5.3.2 Flow Cytometry results for mineral and artesian bottled water samples ... 87

5.3.3 Flow Cytometry results for sparkling bottled water samples ... 92

Chapter 6 : Microbiological Stability Assessment ... 94

6.1 Overview ... 94

6.2 Methods ... 95

6.3 Results and Discussion ... 99

6.3.1 Different types of water have diverse microbial growth responses ... 99

6.3.2 Purified waters remain stable for longer times that mineral waters ... 101

6.3.3 Mineral waters all have diverse microbial growth responses ... 101

6.3.4 Plastic bottles favor microbial growth ... 106

6.3.5 The act of transferring water between bottles was not responsible for the increased microbial growth ... 109

6.3.6 Change in microbial community composition observed through fingerprinting 110 Chapter 7 : Microbial Community Analysis ... 113

7.1 Overview ... 113

7.2 Methods ... 113

7.3 Results and Discussion ... 116

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7.3.2 Microbial community analysis results and most abundant generas ... 119

7.3.3 Distribution of bottled water samples based on microbial communities ... 123

Chapter 8 : Conclusion ... 126

REFERENCES ... 129

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LIST OF ABBREVIATIONS

Al Aluminum

Br Bromide

°C Celsius

Ca Calcium

CFU Colony forming units

Cl Chloride

Cu Copper

DNA Deoxyribonucleic acid

EC European Commission

EPA Environmental Protection Agency

F Fluoride

FCM Flow Cytometry

FDA U.S. Food and Drug Administration

Fe Iron

HCO3- Bicarbonate

HNA High nucleic acid

IBWA International Bottled Water Association IC Ion Chromatography

ICP-OES Inductively Coupled Plasma- Optical Emission Spectroscopy

K Potassium

KAUST King Abdullah University for Science and Technology LNA Low nucleic acid

Mg Magnesium

mg/l Milligram per liter

mL Milliliter Na Sodium Ni Nickel NO2- Nitrite NO3- Nitrate Pb Lead

PCA Principal component analysis PPM Parts per million

RO Reverse Osmosis

SASO Saudi Arabian Standards Organization SO32- Sulfate

TDS Total dissolved solids TOC Total organic carbon WHO World Health Organization

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LIST OF ILLUSTRATIONS

Figure 1.1: Schematic including this study’s chapters along with a brief description of the

experimental procedures. ... 19

Figure 2.1: Global bottled water market share by product (%) in 2020. 34 ... 20

Figure 2.2: Desalination plants located in Saudi Arabia 49. ... 25

Figure 2.3: Purified bottled water production steps (Nestle) 44. ... 27

Figure 2.4: Percent growth of global water consumption and extractions along with population growth 30. ... 28

Figure 2.5: Time in years for freshwater resource renewal 54 ... 29

Figure 2.6: The state of global aquifers depletion54 ... 29

Figure 2.7: Differences between HPC and FCM methods for microbial analysis 84 ... 37

Figure 3.1: Map of distinct water bottle origin locations with the number of brands found in each country. ... 48

Figure 3.2: Percent difference between the average prices of the different types of water. ... 54

Figure 4.1: pH values of drinking bottled water samples as noted on the bottle label with experimental pH values measured using a pH meter annotated in the chart. ... 67

Figure 4.2: Distribution of different types of bottled waters based on mineral composition. ... 81

Figure 5.1: Average microbial count (cells/mL) of total and live cells found in all bottled drinking water samples as determined through FCM. ... 85

Figure 5.2: Average microbial count (cells/mL) of total and live cells found in purified drinking water types as determined through FCM. “n” refers to the number of experimental replicates. “Status” indicates the total microbial cells/mL and live cells/mL. ... 87

Figure 5.3: Average microbial count (cells/mL) of total and live cells found in natural mineral water as determined through FCM. ... 89

Figure 5.4: Average microbial count (cells/mL) of total and live cells found in natural artesian water as determined through FCM. ... 89

Figure 5.5: Variations in microbial cell counts between Evian in glass and in plastic. .... 91

Figure 5.6: Average microbial count (cells/mL) of total and live cells found in sparkling water types as determined through FCM. ... 92

Figure 6.1: Experimental setup of the online FCM system and bottled water incubation.97 Figure 6.2: Design of transplantation experiment using Evian plastic and glass bottles. . 98

Figure 6.3: Incubation experiments to test for microbiological stability over time for various types of bottled waters. ... 103

Figure 6.4: Comparison of growth during incubation in plastic and glass bottles by transferring of Evian water from glass to plastic and vice versa. ... 108

Figure 6.5: Changes in microbial fingerprints and community composition of bottled water samples during incubation. ... 111

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LIST OF TABLES

Table 3.1: General information noted of each drinking bottled water found during

surveying on 09/22/20. The asterisk symbol denotes undisclosed details. ... 41 Table 3.2: Miscellaneous distinctions among some brands that have more than one type of bottle material, type of water, or source location. ... 45 Table 3.3: Brand distributions of types of water, along with general source countries and type of bottle materials. For the entirety of the study, the following four types of water corresponded to the assigned bottled water brands. ... 46 Table 3.4: List of bottled water samples which had been purchased on 26-27/10/20, along with price per mL in Saudi Riyals currency (SR), and production/expiration dates. ... 50 Table 4.1: Average composition values of elements, total dissolved solids (TDS), total hardness, and pH as listed on each bottled water label. ... 58 Table 4.2: Reagents and standards concentrations used for ICP-OES experiment. ... 62 Table 4.3: Range of composition values for each type of water. The name of the brand sample was noted beside the value if no other samples of the same water type contained information for the same element. ... 64 Table 4.4: Anionic composition of bottled water samples from labels compared with experimental results obtained through ion chromatography. ... 69 Table 4.5: Label and experimental values of ions and metals present in bottled water samples, and the percent difference between the two. ... 72 Table 6.1: Details of bottled water samples used for each incubation experiment

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Chapter 1 : Introduction

Water is an essential resource for all living beings. It is used in many important

industries, in agriculture, and for personal consumption 1. Over 70% of the earth’s surface is covered in water, but about 3% of that volume consists of fresh water, while even less is suitable for direct drinking 2. Advancements in water treatment, such as seawater desalination and disinfection, and groundwater pumping or retrieval from wells have allowed water to be more readily accessible 1. Municipal water distribution systems have also aided in accessibility as it can now be delivered directly to the taps of individual consumers 1.

One of the most common forms of water distribution is through packaging it as bottled water. The bottled water industry has grown continuously over the past few decades 2. In 2019, bottled water accounted for $292.9 billion in worldwide revenue for non alcoholic beverages, and revenue has been increasing at an annual rate of 7% since 2012 3. Bottled drinking water has proven to be beneficial during emergency situations, in remote locations, in areas with poor water quality 4, 5, or in places without proper water

distribution infrastructures 6; during emergencies or while in countries lacking access to clean water or proper distribution systems, water may be subjected to pollution, waste hazards, harmful microbes or toxins 7. However, even in many developed countries with improved tap water quality that regularly undergoes rigorous quality control checks, there remains a continued growth in bottled water consumption 8.

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or purified water produced from treatment processes like distillation or reverse osmosis 9.

Nonetheless, potable water is a clean substance able to sustain life and satisfy our survival needs 10. On that account, why are there countless brands of different kinds of bottled water being marketed to people and what is the distinction between them?

According to several studies and surveys, consumers preferred bottled water over tap due to taste preferences and perceptions of the quality being better and healthier due to the water’s source or mineral content, and for this reason there exists specific favoritism towards certain brands or types of water 11. In addition, the increase in advertisements and publicity of drinking bottled water has lead to increase in demand due to the belief in its claims of purity, health benefits, trust in its quality and even the brand’s image 11, 12. For this purpose, natural mineral water extracted from remote and pristine areas has grown in popularity 13. However, there is no evidence that any specific type of bottled water is inherently better, or that bottled water has better quality than tap water 11, 14. In fact, tap water regulation is often more stringent than that of bottled drinking water, which in most cases is regulated as a food item 15, 16. Also, purified bottled water may even originate from the municipal water supplies, essentially making it bottled tap water sold at a higher price 13, 17.

There are several factors to consider regarding the increased availability of different bottled waters, such as the variation in mineral content and the microbiological quality of the water. Depending on the source, compositions and properties of each water category may vary 18, 19. Yet, the presence of minerals in bottled waters may not necessarily be

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minerals present, as some studies have shown, may even contradict its label and the limits prescribed by health organizations, and health concerns may arise from increased trace elements exposure 22231924. In terms of microbial composition, it is often assumed that bottled water is superior and less contaminated, even though the microbiological quality is not disclosed on the bottles 8, 15. However, some studies have shown that there was no significant difference between the quality of tap water and bottled water 25, some have

even shown contamination present in bottled waters 26, and some have shown that storage time and temperature can also affect the microbial composition of bottled water more than tap water 15. Purified waters, often produced via reverse osmosis and disinfection 19, are mostly free of microbial cells and contaminants due to the advanced filtration and disinfection 27, however, natural mineral waters do not go through any treatment and are

able to maintain their natural microbial flora 19. Although microbial quality is regulated in bottled waters 28, there may be implications concerning the microbiological stability in

bottled water, which can lead to the deterioration of water quality 15.

Along with considerations of water quality, bottled drinking water can lead to negative impacts on the environment, and depending on the origin of the water, the costs can be high. Apart from the growing plastic pollution that occurs due to non-degradable

materials, the energy required and the resulting carbon footprint to produce bottled water is tremendous, and the energy to transport it is even greater 17. Greenhouse gas emissions are a direct effect of burning fossil fuels resultant from plastic production and increasing distances of transportation. Importing bottled water can be uneconomical and

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water from natural resources for the production of bottled natural mineral or artesian water can lead to significant environmental consequences as well 29. Ramifications of excessive water withdrawals, from natural resources at a greater rate than can be replenished, may include depletion of fresh water supplies for the local community and agriculture, drop-in aquifers which cause adverse effects and changes in its

characteristics, wells drying up and risk of salt water intrusion, and impacting local wildlife and vegetation which depend on fresh water availability 29, 30.

Saudi Arabia is an arid country that has limited freshwater resources 31. Drinking water is provided through nonrenewable depleting groundwater and desalinated water 7. Saudi

Arabia is also the largest, and growing, desalinated water producer in the world, and the quality of produced water, which is also the same quality of tap water, is safe enough for consumption 25. Locally produced purified bottled water has been found to be compliant with health organizations’ standards just as well as imported water 7, 32, whereas imported

bottled waters have been shown in some cases to exceed health standard limits for some minerals more than local bottled waters 33. Because the quality of locally produced purified bottled waters is up to health standards, it serves the primary basic function that drinking water should, and depending on it, can alleviate the environmental impact associated with transportation and fuel emissions through importing from out of the country 17.

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sources, analyzed for physical-chemical components to corroborate the composition labels, and measured for bacterial quality to examine the microbiological stability.

Although flavored water and vitamin-enriched water have recently grown in popularity 2, they were not analyzed for the purpose of this study due to the nature of additives.

Through various experimentation methods, we will attempt to answer the following questions:

 How many types of bottled drinking waters are available? How many are locally sourced or imported? Is the inventory partial to one type or source?

 Do the composition labels accurately reflect the physical-chemical values of the corresponding bottled waters based on experimental data? Does it adhere to global health organizations’ drinking water guidelines?

 How does the microbial quality differ between bottled water brands depending on the source?

 How stable is the microbiological quality of bottled drinking water?  Is there a difference in microbial communities with respect to the source of

bottled drinking water?

 Based on the results, is there an advantage to drinking natural mineral waters as opposed to purified drinking water?

To answer the questions above, this study was organized in the course of action as can be observed in the schematic in Figure 1.1. The proposed analysis of various bottled

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stability, and if these attributes correspond to its monetary value and status. Additionally, if there is no apparent difference or benefit to drinking imported or bottled natural waters, this study may aid in supporting and encouraging the use of more sustainably sourced and local bottled waters, which will alleviate negative environmental and economic

consequences associated.

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Chapter 2 : Literature Review 2.1 The bottled drinking water industry

Over the years, the bottled water market has grown continuously worldwide, especially in countries with clean tap water provided at little to no cost 10. Global revenue has been estimated to increase at an annual rate of 7% between 2012 and 2025, while volume sales increase by 3.6% 3. Volumes of bottled water sold are expected to surpass 500 billion

liters worldwide by the year 2024. In 2019, western Europe was found to be the most avid consumer, however, Germany and the United States had the highest annual revenue per capita at US$213.1 and US$206.7, respectively 3. There are several key players leading the bottled water market. The top brands globally include Nestlé, which include labels such as San Pellegrino, Vittel, and Nestlé Pure Life; Danone, which includes Volvic, and Evian; the Coca-Cola Company; PepsiCo; Voss water; and Fiji water

company 3, 34. Although PepsiCo and the Coca-Cola Company are leading corporations in

the carbonated beverage market, the increasing demand for bottled waters has driven them to join this market as well 35. The types of water that dominate the bottled water industry are purified water, which held over 35% of the market share in 2020, followed by mineral water (Figure 2.1) 34.

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There are various reasons that can be attributed to the success of the bottled water industry, including marketing and consumer preferences. Retail and advertisements of bottled water have profited off of concerns regarding tap water quality, and global health and wellness trends 3, 36. Campaigns often emphasize the purity and health of water10, 11, and some may even directly highlight issues with tap water contamination to promote their water as a safer replacement 11. Images of pristine water sources such as mountains

and springs may be placed on the bottle label to further emphasize that the water is pure and of good quality 10, 11. Consumers have been led to believe that bottled water is purer,

healthier, and safer from waterborne illnesses than tap water18. Marketing may also be used to link bottled water consumption with physicality, exercise, and health 36. Consumers may prefer replacing other drinks with water, which has no calories or artificial ingredients 37. The increased awareness over links between increased soda consumption and obesity and poor health have also led to an expansion in bottled water usage2. In some alternative cases, marketing may not necessarily be associated with health, but with trendiness and style; Perrier was promoted as a status symbol, and Dasani was advertised as a lifestyle drink 11. Bottled mineral waters, which are sold for billions

of dollars annually worldwide, have been marketed successfully as a better quality drinking water 38. In Europe, mineral springs were often thought to have medicinal

properties and sometimes religious healing powers due to their unique chemical

compositions 10, 38. Historically, mineral waters were used as bathing sites and associated with relaxation, healing, and health. After regulatory implementations and

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regulated by the government as a public good or right, so it is often not marketed, and its price and distribution are controlled 39.

Aside from marketing and health concerns, consumers may prefer bottled over tap for other reasons. Organoleptic properties, such as taste, may encourage bottled water

preference if tap water is chlorinated, for example. Indeed, one study found that there can be as many as thirteen flavors of drinking water40. Some consumers may also prefer

mineral or spring water 11. Bottled water has proven to be more convenient than tap water in some instances. Water can be packaged in different containers, such as glass or

plastic41. It is portable and can be transported and stored while maintaining its quality

over longer periods of time, most importantly during natural disasters or emergency situations where municipal water can be susceptible to contamination. Urbanization and change in working environments have also aided in bottled water growth18. The

availability of bulk and single use packaging, and the low cost of bottled water in comparison to other beverages, may also contribute to bottled water consumption 37.

2.1.1 Classifications of bottled water

There are several categories of bottled waters based on their source, mineral composition, or treatment. According to the International Bottled Water Association (IBWA), types of bottled water include 42:

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reverse osmosis, 1 micron filtration, and Ozonation 43. Water is re-mineralized after

treatment 44.

Well water: Underground water from a drilled or bored hole created in the ground to extract water from the aquifer 42.

Spring water: Underground water from an aquifer that naturally flows to the surface of the spring location. It may also be extracted, without altering the natural quality of the water, through a borehole reaching the underground water which feeds into the spring 42. Spring water does not undergo any further treatment and its mineral content may vary based on the spring 18.

Artesian water: Water in a well trapped aquifer where the level of the well water is above the level of the aquifer 42. The naturally created pressure from digging a hole into the

aquifer pushes the water out 43.

Sparkling water: Carbonated water, which often loses its natural carbonation after

emerging from the source. However, bottled sparkling water contains the same amount of carbon dioxide that it contained naturally from the source after treatment and possible replenishment of carbon dioxide. This type of water may be labeled as sparkling mineral, spring, or drinking water 42.

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they are meant to retain their microbiological and natural states, however, they may undergo replenishment of carbon dioxide if the water was meant to be naturally carbonated and lost some carbonation during extraction 38.

2.1.2 Drinking water in Saudi Arabia

Saudi Arabia is one of the driest countries in the world lacking natural water resources. The United Nations has classified Saudi Arabia, as well as other Middle Eastern countries, as water-scarce nations 45. Paradoxically, it also has one of the highest water consumption rates in the world 46, 47. In 2019, the amount of water consumed daily

reached 263 liters per capita 47. The water demand has been increasing by 8.8% percent annually 45, and demand is expected to grow up to 25.79 billion m3 by 2025 47. Sources of surface water and renewable groundwater are severely limited due to low levels of

rainfall and high rates of evaporation 45, 46. Other sources of water that are used to meet the country’s water demands include nonrenewable groundwater sources, from

sedimentary and deep rock aquifers, and desalinated seawater 45. Groundwater reserves

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desalinated water production for decades, representing 22% of global production and 7.3 million m3 daily 45, 47. The majority of desalinated water is produced through thermal desalination, which consists of multi-stage flash (MSF) or multi-effect distillation

processes (MED), while membrane desalination via reverse osmosis (RO) produces about 20% of desalinated water 19, 48 (Figure 2.2). Regardless of the method used, desalination

stages include water feed intake, pretreatment, desalination which removes salts, and post-treatment that involves disinfection and re-mineralization 48. Desalination has been recognized to provide high quality and potentially unlimited drinking water abiding by health regulations while preserving natural freshwater supplies 46, 48, 49.

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In the matter of bottled drinking water, Saudi Arabia claimed the spot for the twelfth highest bottled water consumer per capita with over 31.7 gallons per resident in 2019 37. Despite the production of good quality drinking water through desalination, many consumers prefer drinking from bottled waters due to assumptions of poor tap water quality 50. In 2013, it was estimated that nearly 52% of the 183 bottled water companies located in Gulf Cooperation Council (GCC) countries belonged to Saudi Arabia 41, 51.

There were also over 90 bottled drinking water manufacturers noted recently in the country 52. In general, the bottled water market in the Middle East, which is dominated by

Saudi Arabia, is expecting a compound annual growth rate of 6.2% between 2019- 2024 and reaching a value of over 145 billion USD by 2024 50.

Purified bottled waters are produced in a process similar to desalination (Figure 2.3). Water undergoes pretreatment, demineralization, filtration, and post treatment including disinfection and re-mineralization to the brand’s standards 44. In some cases, the bottled

water is sourced from municipal supplies, and then goes through an extra separate

purification process. The quality of purified bottled water is often found to be no different than tap water produced by desalination 41, 53; the main distinction is the form of

distribution. There are also several bottling water companies in Saudi Arabia that receive their water from both underground and desalination 41; this indicates that the bottled

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Figure 2.3: Purified bottled water production steps (Nestle) 44. 2.2 Environmental impact of bottled water

Consuming bottled drinking water can lead to various negative environmental

consequences. Bottled water may place stress on local water resources, as bottling natural water may lead to a depletion of nonrenewable freshwater obtained from the surface or underground 54. There has been a significant increase in water withdrawals and

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Figure 2.4: Percent growth of global water consumption and extractions along with population growth 30.

Groundwater renewability depends on several environmental factors, but may take

several hundreds to thousands of years to replenish (Figure 2.5). Only 6% of groundwater reserves are renewable within a human lifetime 54. One NASA study showed that 21 of the largest 37 aquifers in the world are being depleted at a rate faster than they are being replenished, with the Arabian aquifer system experiencing the largest stress 55 (Figure 2.6). Groundwater also depends on fresh surface water to feed into it and vice versa, and the reduction in surface water reserves affects groundwater in return. Increased water intake, in conjunction with pollution and climate change, affect the quality and amount of freshwater availability. Over-exploitation of natural water resources also threaten the livelihood of wildlife and plants through risks of salt water intrusion and reduction in the water level, which can change the physical and biological characteristics of the

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Figure 2.5: Time in years for freshwater resource renewal 54

Figure 2.6: The state of global aquifers depletion54

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The resultant energy from transportation depends on the distance from the bottling plant and the vehicle(s) used to deliver to the market 17. Demonstrating this impact through a calculated scenario, purified local water transported within Los Angeles via truck costs about 1.4 mega joules of energy per liter, while French spring water exported across the ocean to Los Angeles costs over four times more energy through several modes of transportation 17. Greenhouse gas emissions are a direct effect of burning fossil fuels

which results from plastic production for bottle materials and increasing distances of transportation. These harmful emissions contribute to the effects of climate change and pollution, and can consequently lead to issues involving the few freshwater resources available. Although Saudi Arabia is an arid and water scarce country, importing bottled waters sourced from countries with natural freshwater does not necessarily signify better water quality. Several studies have analyzed domestic and imported bottled water brands in Saudi Arabia and found that the quality of both were comparable and compliant with health regulations for drinking water 31, 57.

2.3 Regulatory Requirements

Regulations on bottled drinking water standards vary by region, and each country has its own standards for monitoring drinking water quality 58. In Saudi Arabia, the Saudi Arabian Standards Organization (SASO) oversees bottled water regulations32. In the

USA, the U.S. Food and Drug Administration (FDA) is responsible 16. However, the

World Health Organization’s (WHO) guidelines for drinking water quality define recommended criteria at an international level and acts as an ideal guide for national standards and safety 44, 58. Many organizations take into consideration the WHO

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water quality is strictly regulated by the European Commission (EC) under the guidance and recommendations of the WHO 44, 60. Nonetheless, WHO guidelines are followed while taking into account local conditions, financial and technical capabilities, and environments since not all parameters are relevant to some regions 58. Therefore, there is no universal path to drinking water regulation 58. Organizations that monitor bottled drinking water may also differ from those that monitor municipal water quality, and standards set may not be equivalent. In the USA, for example, the Environmental

Protection Agency (EPA) manages regulating municipal water, which is stricter than the FDA that categorizes bottled water as a food item 16. The WHO provides a

comprehensive guideline and presents risk assessments of the numerous microbiological, inorganic and organic chemical, radiological, and physical pollutants that may be found in drinking water 59. They also derive recommended concentration value limits for these hazardous elements where relevant. Guidelines are also set for parameters that may affect the acceptability of water’s taste, odor, or appearance 59.

2.4 Mineral composition of bottled water

Bottled drinking water possesses a composition label with the amount of minerals, salts, and ions present. Bottled water, and tap water, typically includes minerals such as

calcium, magnesium, potassium, sodium, and chloride, which can be present as inorganic salts 61. These minerals are found naturally in water but can be added to purified water in

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quantities may lead to adverse health consequences and affect metabolism, intake of essential nutrients from food, functions of the intestinal mucous membrane, or other functions 62.

Concentrations of minerals in water vary widely in all natural waters from different parts of the world as water dissolves elements and metals from the earth 38. Classification of mineral waters depend on the natural high amount of dissolved solutes 38. During

production of purified waters, minerals are added post disinfection after water has been de-mineralized during reverse osmosis and filtration 44. WHO guidelines set very low concentration limits for harmful metals and trace elements such as lead, copper, and iron, and larger concentrations are recommended for major nutrients such as calcium and magnesium; however there are no guideline limit values set because there are no health risks associated with the concentrations present in drinking water63.

Many research studies in several countries have examined the elemental and chemical constituents present in different types of bottled water including major cations, anions, pH, and trace elements, while verifying its compliance with the WHO and other health organizations guidelines. Analysis methods included ion chromatography (IC) 25, 31, 38, 64 mainly to study anion composition, and Inductive Couple Plasma Mass Spectrometry (ICP-MS) 7, 38, 52, 65-67, or Inductive Couple Plasma Optical Emission Spectrometry

(ICP-OES) 31, 38, 67-69 for cation and trace metal analysis. Studies indicate that most bottled

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indicated on the manufacturer label 65, mineral water in glass bottles had higher median

concentrations of some elements 38, and that the quality of bottled water may be less than municipal water 67. Studies done in Saudi Arabia also show similar results, 7, 41, 52, 65. One study comparing imported and local bottled waters showed that they all comply with WHO standards, however the measured values show considerable variations from those of the labels and between replicates 31. Other researchers also indicate discrepancies

between experimental and labeled values 70, 71. Another study showed that there is no significant difference between the quality of bottled water and tap water 25. One study

indicated that local bottled waters were more reliable in compliance with standards than brands that were imported 32. Nonetheless, bottled waters, whether mineral or purified, were mostly compliant with health organization guidelines and standards, and may not pose a great risk to the average consumer. There are several studies that disclose the names of the examined bottled water brands 31, 41, 57, 71, however, they do not discuss

details regarding purified or mineral water sources or treatment conditions.

2.5 Microbiological quality and stability of bottled water

2.5.1 Regulations for microbiological composition of bottled water

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Regulations implemented by the WHO and other health organizations monitor the presence of harmful known microbial species. The WHO guidelines identify 43 microbiological parameters that include toxic bacterial, viral, protozoan, and helminth contaminants 59. Some guidelines also indicate limit values for microbiological

parameters, such as total coliforms, total heterotrophic bacteria, or known pathogens such as Escherichia coli (E. coli) and Pseudomonas aeruginosa 59. The WHO guidelines have

specified limits for known pathogens; E. coli must be undetectable in 100 mL of water. However, WHO guidelines do not specify limits for total coliforms or total heterotrophic bacteria as values are not useful for pathogen detection; it only recommends testing these parameters as an indication after treatment or disinfection to assess the adequacy of treatment and distribution while maintaining low microbial numbers 59, 63. On the other

hand, guidelines for mineral waters from the European Parliament & the Council of the European Union (Directive 2009/54/EC) indicate that heterotrophic plate counts should be ≤ 100 colony forming units (CFU) per mL at 22°C and ≤ 20 CFU per mL at 37°C 73.

Also, in terms of specific microbial contaminants, E. coli, P.aeruginosa and Enterococci must not be detectable in 250/mL of water 44, 73. Purified bottled waters undergo

membrane filtration, reverse osmosis, and/or disinfection which are effective in removing microbial cells and pathogens as small as 0.2 microns 44. However, natural waters

maintain their microbiological states and cannot undergo extensive treatment or disinfection processes, so microbial species present remain intact after bottling 74.

2.5.2 Microbiological research studies on bottled drinking water

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through heterotrophic plate count (HPC). It is culture based method which is done through filtration of the sample through a filter which retains microbial cells, then incubating it on a nutrient rich medium, such as agar, at a set temperature commonly between 22-37°C 73, 75, 76. The viable cultivable microbial colonies are then counted and represented commonly as CFU 73, 75, 76 or as Most Probable Number (MPN) of

presumptive coliforms 72. Most research has been done for mineral bottled waters 25, 73, 75, 76. However, one study examined spring and purified bottled waters and found that total

coliform count was absent 72. In Saudi Arabia, another study on local bottled waters also

did not determine any CFU and found that bottled waters were comparable in microbiological quality to tap water 25.

Besides the aforementioned studies, most research on mineral water found positive HPC and several coliform contaminated samples. Bottled water may represent oligotrophic environments for bacteria and possess enough nutrients and substrates to sustain

microbial growth 52, 77. The most frequent phylum of bacteria found in drinking water is Proteobacteria78. Contaminants found in some samples included Escherichia coli 73, 76, 79,

Pseudomonas aeruginosa 52, 73, 76, 79, Enterobacter aerogenes43 and Staphylococcus sp. 52,

76. There were several interesting findings among these studies. The studies also show

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potentially pathogenic bacteria were found in a smaller percentage of carbonated samples than still water 73. In terms of microbiological stability, research show that microbial concentrations increase over time after bottling. Mineral waters are known to increase in cell numbers up to104- 105 CFU/mL within one week after bottling 77. One study has also found that imported mineral waters and bottles with older production dates had the highest CFU reaching values of up to 104 CFU/mL 75. This indicates that the microbial

quality of bottled water may be affected by storage time, and perhaps storage conditions which are unknown. This study also found that microbial analysis indicated that nearly 80% of dominant strains identified were gram negative bacteria which constituted Pseudomonas or similar 75. Studies containing a comprehensive analysis of all types of microbial species, apart from contaminants, found in several bottled mineral waters have not been demonstrated.

2.5.3 Determining microbiological content through Flow Cytometry

Upon considering methodologies for evaluating and enumerating microbiological components of water, HPC is not very efficient when it’s compared to newer and more innovative methods of biological content analysis, such as Flow Cytometry (FCM) 80 (Figure 2.7). It has the capability to rapidly calculate the total, as well as intact, cell concentrations through fluorescent DNA staining methods 81. HPC is only able to detect

viable cells and typically less than 1% of total bacteria is cultivable 81. FCM is also able

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Figure 2.7: Differences between HPC and FCM methods for microbial analysis 83

Although FCM has been used extensively on drinking waters to investigate treatment methods, water quality water stability and bacterial growth81, 84-87, relevant studies to determine the microbial quality of a variety of bottled drinking waters have not been established. There are a few studies who have implemented the analysis of a bottled mineral water in their research, commonly Evian, to compare results with other variables

87, 88. One study showed that bottled mineral water had a higher HNA cell percentage than

tap water, but both contained total cell concentrations were valued at 103 -105 cells/mL 82. Other studies analyzed microbial growth potential or limiting factors in water samples including bottled mineral water 80, 87, yet, the mineral water was filtered, transferred into laboratory glassware, or altered in some way. Consequently, the natural microbial quality confined in the bottle as originally presented to the consumer is undetermined. Another study examined groundwater after microfiltration and bottling in a PET container, which would resemble purified water 89. Results illustrated that as much as 10% of native

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observed up to 1-3 × 105 cells/mL after bottling and storage 89. Researchers were able to

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Chapter 3 : Surveying Available Bottled Drinking Water

3.1 Overview

There are numerous bottled drinking water brands available in the market from different origins. However, the amount accessible by the consumer at a given time in one location has not been explored within KAUST. For the first part of this study, we wanted to uncover how many types of bottled waters are present locally, what differences can be noted, and where these bottles are coming from. Uncovering this information can help identify the extent of influence in the local market, and the source of any environmental or economic impact.

3.2 Methods

The primary local supermarket (Tamimi Supermarket at KAUST) was explored on the morning of 09/22/20 to obtain a list of the entire available inventory of bottled water brands. The supermarket was visited during the morning when there would be fewer people expected and the most stock available. For the purposes of this study, vitamin-enhanced or flavored waters were excluded. The label of each bottled water was also analyzed for type of water, origin source, bottling location, and composition (Chapter 4). The assortments of bottled waters were purchased over two consecutive days

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3.3 Results and Discussion

3.3.1 Initial bottled water survey findings

Overall, there were a total of nineteen distinct brands of bottled drinking water discovered in the supermarket (Table 3.1). The labels of each bottle contained similar details. Stated information included the brand name/logo and type of water, volume, average chemical composition values (discussed further in Chapter 4), source of water, bottling location, and production and/or expiration dates (Table 3.4). Some bottles also had additional information about any treatment process underwent, whether it be natural or man-made.

According to the labels, five types of bottled water were identified: bottled drinking water, natural mineral water, spring water, artesian water, and sparkling water. Many of the brands that were labeled as natural mineral waters were sourced from spring water (e.g. Evian and Volvic), hence, the brand labeled as spring water (Wildalp) was

categorized as mineral water throughout the study. Still Scottish mountain water was also categorized as mineral water because it matched the mineral water description 18.

Sparkling water was identified as sparkling natural mineral water (e.g. Badoit), artificially carbonated purified water (e.g. Berain), or artificially carbonated natural mineral water (e.g. Perrier). The water containers were made of either plastic or glass material. Several brands had both plastic and glass bottles for the same type of water (e.g. Berain, Evian, Voss), while other brands were solely found in one type of bottle material; sparkling waters were only contained in glass bottles. Glass retains CO2 better than

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Table 3.1: General information noted of each drinking bottled water found during surveying on 09/22/20. The asterisk symbol denotes undisclosed details.

No. of Brands No. of Distinct Bottles Brand Name Bottle Material Type Of

Water Water Source

Location Of Bottling Type of Treatment Bottle volume (mL) Price (SR) 1 1 Tamimi Markets Bottled Drinking Water

Plastic Purified water

Well water

Hana company for food industries. AlQassim- Buraidah, KSA * 200 0.35 330 0.40 5000 * 2 Well water- Al Shadida Valley Tania bottled drinking water factory. AlKharj, KSA Ozonation 200 0.35 600 0.75 2 3 Nestle Pure

Life Plastic Purified water

Underground water or desalination water

Springs water factory

co. Dammam, KSA *

200 0.60

4 Desalination water Naqiah water factory

co. Madinah, KSA

Reverse Osmosis 330 0.60 1500 1.80 3 5 Berain Plastic Purified water * Berain company. Jeddah, KSA Ozonation 200 0.55 330 0.60

Well water Berain company.

Riyadh, KSA 12000 * 6 Glass 300 2.75 7 Glass Sparkling water 750 7.05

4 8 Hana Water Plastic Purified water Well water

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No. of Brands No. of Distinct Bottles Brand Name Bottle Material Type Of

Water Water Source

Location Of Bottling Type of Treatment Bottle volume (mL) Price (SR)

5 9 Arwa Plastic Purified water Well water

Saudi Coca cola beverage bottling

company. Sudair city, Riyadh,

KSA * 200 0.60 330 0.80 1500 * 6 10 Evian Plastic Natural

mineral water French Alps

Cachat spring- S.A.E.M.E. Evian, France Natural filtration through rocks 330 6.25 500 * 1250 13.40 11 Glass 330 8.00 750 12.00

7 12 Volvic Plastic Natural

mineral water French Volcanoes

Clairvic spring. Volvic, France (bottled at the source) Filtration through 6 layers of volcanic rock 330 5.95 500 6.50 750 8.70 1500 13.70 8 13 Voss Plastic Artesian natural mineral water Artesian water- Norway Artesian water- Norway

Voss production AS. Voss, Vantestrom, Norway (bottled at Voss source) * 330 8.00 500 8.55 14 Glass 375 10.65 800 18.05 15 Glass Sparkling water 375 10.65 9 16 Perrier Glass Natural mineral water & CO2

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No. of Brands No. of Distinct Bottles Brand Name Bottle Material Type Of

Water Water Source

Location Of Bottling Type of Treatment Bottle volume (mL) Price (SR)

10 17 Aquafina Plastic Purified water Groundwater

AlJomaih beverage bottling company. Buraidah, KSA Purified using Aquafina's HydRO-7 process 200 * 330 0.50 11 18 Badoit Glass Sparkling natural mineral water

Saint Galmier, France

SAEME. Saint Galmier, France (source Badoit) * 330 7.30 750 11.20

12 19 Fiji Plastic Natural

artesian water

Yaqara, Viti Levu, Fiji Islands

Natural waters of viti

limited. Yaraqa, Fiji *

500 5.70

13 20 Acqua

Panna Glass

Natural

mineral water Toscana, Italy

Sanpellengrino S.p.A. Acqua Panna

springs, Florence, Italy (bottled at souce) * 250 5.05 14 21 Highland Spring Plastic Natural mineral water

Drawn from organic land in Scotland Highland spring Ltd. Perthshire, Scotland, UK (bottled at source) * 500 3.60 15 22 S. Pellegrino Glass Sparkling natural mineral water

San Pellegrino, Italy

Sanpellegrino S.p.A. San Pellegrino Terme, localita Ruspino, Italy (bottled at source) * 250 4.33 (6 pack for 25.95) 16 23

Nova Plastic Purified water

Underground well water-Saad City

Health water bottling company. Saad, KSA * 200 0.55 24 Well water- Nufoud Se'ed HWB co. plant. Seed, KSA 330 0.60

17 25 Wildalp Plastic Natural

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No. of Brands No. of Distinct Bottles Brand Name Bottle Material Type Of

Water Water Source

Location Of Bottling Type of Treatment Bottle volume (mL) Price (SR) 18 26 Still Scottish Mountain Water by Sainsbury's Plastic Still mountain water, underground Caledonian water- eastern edge of the

Campsie Fells Produced in the UK for Sainsbury's Supermarkets Ltd, London Natural filtration through layers of volcanic rocks 500 6.50

19 27 Tannourine Plastic Mineral water Natural spring water from

Tannourine Lebanon

bottled at the source by Societe Libanaise des Souces des Eaux de Tannourine s.a.l Lebanon (Daher &

Sarkis Sources)

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Studying the labels for water sources and bottling locations unintentionally brought about some curious revelations. It was uncovered that some brands had bottles with water that was sourced or bottled at different locations. Table 3.2 indicates that Tamimi Markets, Nestle Pure Life, and Nova all contained bottles that were sourced from different cities in Saudi Arabia. Also, certain Tamimi Markets bottled drinking water was bottled at the same location as Hana, which can be noted in Table 3.1. Specific bottled water brands containing samples with more than one bottling location or bottle material (plastic/glass) shall be identified throughout the rest of the thesis as: Brand name- location or brand name- bottle material.

Table 3.2: Miscellaneous distinctions among some brands that have more than one type of bottle material, type of water, or source location.

Brands with Variations Among Bottled

Drinking Waters Distinct Features Present

Berain Two types of water, three types of bottles: Purified (glass/plastic), sparkling Voss

Two types of water, three types of bottles: Artesian natural mineral (glass/plastic),

sparkling

Evian Two types of bottles:

Natural mineral water (glass/plastic) Tamimi Market Bottled Drinking Water Two water sources with separate bottling

locations

Nestle Pure Life Two water sources with separate bottling locations

Nova Two water sources with separate bottling locations

3.3.2 Origin locations of different bottled drinking waters

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Arabia, and the water originated from desalinated water, well water, or groundwater (Table 3.3). Only one brand was identified as sourced from desalinated water (Nestle Pure Life). Another brand (Aquafina) was treated through reverse osmosis, which is the same process used for desalination. The majority of purified water samples originated from groundwater or well water. Not all specific sources were identified, but the bottling locations varied among several cities in Saudi Arabia. On the other hand, all other types of water were imported from other countries. Most of the waters arrived from Europe, and only one came from Fiji. All mineral waters were bottled at their respective sources, and incidentally, all of these brands are named after the source location.

Table 3.3: Brand distributions of types of water, along with general source countries and type of bottle materials. For the entirety of the study, the following four types of water corresponded to the assigned bottled water brands.

Type of Bottled Water No. of Brands Brands Country of

Origin No. of Sources

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Sparkling water 5 1. Perrier 2. Badoit 3. S. Pellegrino 4. Voss 5. Berain -France -France -Italy -Norway -Saudi Arabia Each bottled at distinct sources of origin (no specific source stated) Glass Artesian water 2 1. Fiji 2. Voss -Fiji -Norway Each bottled at distinct sources of origin (no specific source stated)

Plastic/ glass

Of the nineteen bottled drinking water brands identified in total, only seven originated locally from Saudi Arabia, as noted in Figure 3.1. This reveals that over 60% of the bottled waters were imported, which further supports the notion of environmental impact that importing water bottles may contribute. Purified bottled waters are often produced and packaged close to the local market, whereas natural waters are usually packaged at their unique sources and transported over long distances to places of demand 17. The extent of transportation has a large impact on CO2 greenhouse gas emissions, thus,

transporting water bottles is a burden on climate change issues. Evian transported from France is about 5,000 km away, while Fiji must travel over 15,000 km to reach

consumers in Saudi Arabia. Regardless of the container, the weight of 1 liter of water is about 1kg, and the weight becomes greater when adding on the weight of the bottle. Glass poses a greater impact to the environment, as it is about 40 times heavier for glass than for plastic, and the heavier load results in higher costs in packaging, fuel, and carbon emissions. Transportation of local water via trucks from a distance of 500 km can

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Figure 3.1: Map of distinct water bottle origin locations with the number of brands found in each country.

3.3.3 Bottled drinking water shelf life

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range for both plastic and glass bottles. Although water itself can have an infinite shelf life, no matter the source, water quality and taste can change over time due to chemicals leaching from containers or due to long storage time 16, which suggests that the

production and expiration dates pertain to the packaging.

Also, due to the porosity of plastic bottles 90, environmental elements may leach into the water and affect taste, smell, or acceptability. The FDA, which regulates bottled drinking water in the USA, does not require an expiration date to be listed so long that it is

produced following Current Good Manufacturing Practice (CGMP) quality regulations and is stored properly 16. However, it recommends a two-year shelf life for still bottled

drinking water. The EC, which regulates bottled water in Europe, also recommends a two two-year shelf life for still drinking water, but it also informs that the type of packaging and storage conditions may affect the “best before” date 91. The purified bottled water

samples examined all had an expiration date one year post production. However, they were all sourced from Saudi Arabia, where bottled water is regulated by the Saudi Arabian Standards Organization (SASO) 92, thus, the recommended shelf life and expiration dates may vary.

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Table 3.4: List of bottled water samples which had been purchased on 26-27/10/20, along with price per mL in Saudi Riyals currency (SR), and production/expiration dates.

Brand Distinct Bottles Brand name Bottle Type Type of

Water Water Source

Location of Bottling Bottle Volume (mL) Price per mL Production Date Expiration Date 1 1 Tamimi markets bottled drinking water plastic Purified water Well water

Hana company for food industries. AlQassim- Buraidah, KSA 200 0.0018 22/03/2020 22/03/2021 2 Well water- Al Shadida Valley Tania bottled driniking water factory. AlKharj, KSA 200 0.0018 3/7/2019 3/7/2020 2 3 Nestle pure life plastic Purified water Underground water or desalination water Springs water factory co. Dammam, KSA 200 0.0030 17/08/2020 16/08/2021 4 Desalination water Naqiah water factory co. Madinah, KSA 330 0.0018 16/09/2020 15/09/2021 3 5 Berain plastic Purified water * Berain company. Jeddah, KSA 330 0.0018 22/05/2020 22/05/2021

6 glass Well water Berain company.

Riyadh, KSA 300 0.0092 10/3/2020 10/3/2021

7 glass Sparkling

water Well water

Berain company.

Riyadh, KSA 750 0.0094 23/12/2019 23/12/2020

4 8 Hana water plastic Purified

water Well water

Hana company for food industries.

AlQassim- Buraidah, KSA

330 0.0018 30/12/2019 30/12/2020

5 9 Arwa plastic Purified

water Well water

Saudi Coca cola beverage bottling

company. Sudair city, Riyadh, KSA

200 0.0030 18/03/2020 18/03/2021

6 10 Aquafina plastic Purified

water Groundwater

AlJomaih beverage bottling company.

Buraidah, KSA

330 0.0015 28/07/2020 27/07/2021

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Brand Distinct Bottles Brand name Bottle Type Type of

Water Water Source

Location of Bottling Bottle Volume (mL) Price per mL Production Date Expiration Date

water Nufoud Se'ed Seed, KSA

8 12 Evian plastic Natural mineral water French alps Cachat spring- S.A.E.M.E. Evian, France 330 0.0189 1/10/2019 1/10/2021 13 glass 330 0.0242 17/07/2019 17/07/2021 9 14 Volvic plastic Natural mineral water French volcanoes Clairvic spring. Volvic, France (bottled at the source) 330 0.0180 21/10/2019 21/10/2021 10 15 VOSS plastic Artesian natural mineral water Artesian water from Norway Voss production AS. Voss, Vantestrom, Norway (bottled at Voss source) 330 0.0242 19/12/2019 19/12/2021 16 glass 375 0.0284 11/7/2019 11/7/2021 17 glass Sparkling water 375 0.0284 28/11/2019 28/11/2021 11 18 Perrier glass Natural mineral water & CO2 Water captured at the source in France N.W.S, SUD. Vergeze, France (bottled at source Perrier) 330 0.0183 27/05/2020 1/5/2022 12 19 Badoit glass Sparkling natural mineral water Saint Galmier, France SAEME. Saint Galmier, France (source Badoit) 330 0.0221 20/11/2018 19/11/2020 13 20 Fiji plastic Natural artesian water

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Brand Distinct Bottles Brand name Bottle Type Type of

Water Water Source

Location of Bottling Bottle Volume (mL) Price per mL Production Date Expiration Date mineral water Pellegrino Terme, localita Ruspino, Italy (bottled at source) 17 24 Wildalp plastic Natural spring water still Hochschwab massif spring water

from Styrian Alps/ Seisensteinquelle, Wildalpen, Austria Wildalpen Wasserverwertungs GmbH Sausenbach, Wildalpen, Austria 500 0.0091 5/3/2019 5/3/2021 18 25 Still Scottish Mountain Water By Sainsbury's plastic Still mountain water, under-ground Caledonian water from eastern edge of the Campsie Fells Produced in the UK for Sainsbury's Supermarkets Ltd, London 500 0.0130 * 7/1/2022

19 26 Tannourine plastic Mineral

water Natural spring water from Tannourine Lebanon bottled at the source by Societe Libanaise des Souces des Eaux de

Tannourine s.a.l Lebanon (Daher & Sarkis

Sources)

330 0.0053 9/3/2020 9/3/2021

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3.3.4 Price variations among different bottled water samples

The price of each bottled water purchased was listed in Table 3.4. The prices were affected by several factors including bottle material and type of water. Glass bottles were more expensive than their plastic bottle counterparts; Berain in glass costs over 400% greater per mL than in plastic, Evian costs 28% greater in glass, and Voss costs 17% more. In terms of type of water, as shown in figure 3.2, purified water was the most affordable type of water, on average. The average value of mineral water exceeded that of purified water by over 400%, while artesian water exceeded it by over 600%. The

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Figure 3.2: Percent difference between the average prices of the different types of water.

3.3.5 Study implications

An interesting phenomenon observed throughout the duration of the study was the appearance and disappearance of bottled water brands locally. During the first day of shopping, two brands (Wildalp and Still Scottish) were discovered that were not noted on the day of the first survey, and a third new brand (Tannourine) was found on the second day of shopping that hadn’t been seen previously. Over time, other different brands were observed in the supermarket and in other locations in the community, such as restaurants. There may be analysis limitations due to the continuously changing assortments of bottled water and the circumstances of the enclosed residential campus community, however, the amount of bottled waters obtained suffice to represent the intention of this study. In short, there are obvious variations between local and imported brands of bottled drinking water. Because Saudi Arabia comprises mainly desert land with no natural springs and scarce natural sources of water, there are no available bottled natural mineral

0.0027 0.0145 0.0213 0.0193 0 0.005 0.01 0.015 0.02 0.025 Purified bottled drinking water Natural mineral water

Artesian water Sparkling water SR/mL

Type of Bottled Water

Average Price of Types of Water and Percent Increase Compared to Purified Drinking Water

+ 428%

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waters. However, this does not necessarily affect or diminish the quality of local bottled waters in comparison; Some mineral waters have been found to contain toxic compounds or organic contaminants present due to possible pollution in the aquifers 68. In the

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Chapter 4 : Physical Chemical Analysis of Composition Label

4.1 Overview

All bottled drinking waters possess a label containing the average composition of several ions and minerals, pH values, and total dissolved solids. In spite of the regulations set in place, there are several experimental studies that have proven the inaccuracy of label values 22, 94-96. In the second part of this study, we have explored the physical-chemical compositions of all bottled drinking water samples obtained, compared the values provided on the label, and then conducted several experiments to evaluate its validity on behalf of pH and most listed ions while reviewing its compliance to drinking water health guidelines.

4.2 Methods

The values of the average composition of chemicals, minerals, and pH were obtained from the labels of each bottled water sample and recorded into Table 4.1. In order to evaluate the values noted from the composition labels, several experimental procedures were performed. The samples were analyzed during the same week after purchasing and were stored at room temperature (21-22°C) with their respective caps kept on. For this study, certain uncommon elements, as well as total dissolved solids (TDS) and total hardness were not analyzed. Results were analyzed using Microsoft Office Excel, and the PCA (Figure 4.2) was processed using RStudio with the “FactoMineR” package.

A digital pH meter was used to measure the pH of each sample. The WTW™

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pH values of 4.00, 7.00 and 10.00, at 21°C. The electrode was rinsed with Milli-Q water before use and in between measuring each standard or sample. The electrode was

submerged into each sample at room temperature (between 21-23°C) to obtain the pH reading. The first value the reading stabilized at was the value recorded. However for sparkling water samples, the value promptly recorded was the first to become stable for a couple of seconds. This allowed the capture of the most accurate pH value before the escape of CO2 after opening the bottle; reduction of CO2 concentration would increase

the pH of sparkling water. The electrode was stored with a protective cap containing storage solution before and after use.

Ion Chromatography (IC) was used to measure anions including fluoride (F), chloride (Cl), bromide (Br), nitrate (NO3-), nitrite (NO2-), sulfate (SO42-), and phosphate (PO₄³⁻)

present in each sample. IC functions by separating ions at different rates, or retention times, based on their charge and affinity to the stationary phase in a separation column while the mobile phase aids in transporting the sample through the column. The sample passes through a suppressor which enhances the ion detection before passing through a detector that measures the change in conductivity then translates results into a

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analyze the data. Through the chromatograms generated, the anion concentrations are determined by comparing the area under each peak to those of the standards with known concentrations. The drinking water samples were filtered through 0.22µm syringe filters and were not diluted prior to use.

Table 4.1: Average composition values of elements, total dissolved solids (TDS), total hardness, and pH as listed on each bottled water label.

Average Composition (mg/L)

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Inductively coupled plasma optical emission spectroscopy (ICP-OES) was used to calculate concentrations of most cations indicated in the composition labels and analyze some trace metals. ICP-OES is an analytic technique that determines the chemical composition using a spectrometer and plasma. Ions become excited into a higher energy state through the high temperature inductive plasma (argon gas) and then the ions release radiation at a specific wavelength when they lose this excitation energy. This is

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performed between every ten samples using an ICP multi-element standard (CPAchem) with concentrations of one and ten parts per million (ppm).

Table 4.2: Reagents and standards concentrations used for ICP-OES experiment.

Elements Standards used Concentrations

(ppm) Wavelength analyzed (nm) Na TraceCERT 0.2, 2, 20, 200 568.821 Ca Inorganic Ventures 0.2, 2, 20, 200 315.887 K TraceCERT 0.1, 1, 10, 100 766.491 Mg TraceCERT 0.1, 1, 10, 100 279.800 Fe Inorganic Ventures 0.05, 0.5, 5, 50 234.350 Pb BDH 0.05, 0.5, 5, 50 220.353 Al TraceCERT 0.05, 0.5, 5, 50 237.312 Cu Ricca Chemical Company 0.05, 0.5, 5, 50 327.395 Ni Inorganic Ventures 0.05, 0.5, 5, 50 216.555 Zn Ricca Chemical Company 0.05, 0.5, 5, 50 334.502

The results of ICP-OES and IC were measured in ppm, and were then compared to bottled water label values in mg/L, which is an equivalent measurement.

4.3 Results and Discussion

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composition label; this may indicate that the water did not contain that particular component or that the information was not disclosed.

Minerals listed in all types of bottled water included: bicarbonate (HCO3-), sulfate(SO42-),

chloride (Cl-), fluoride (F-), nitrate (NO

3-), calcium(Ca), magnesium (Mg), sodium (Na),

and potassium (K). Elements that were only unique to certain water types or excluded include: silica (SiO2): not listed on any purified water bottled, bromate (BrO3-): only

listed in purified water, nitrite (NO2-): only noted in one mineral water bottle (Wildalp),

carbonate (CO2-): not listed in any mineral waters, and iron (Fe): not listed in artesian and sparkling waters. With the exception of silica, the constituents that were not listed on every type of bottled water were expected to be present in trace amounts that were not significant. The concentrations of fluoride and nitrate were similar across all types of water.

References

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