This study investigates the caffeine content in fourteen different local carbonated and energy drink brands, utilizing UV spectrophotometry for rapid and accurate caffeine quantification. The results reveal a significant variation in caffeine concentrations across the samples, with the energy drink XL(I) exhibiting the highest concentration at 520 µg/ml, while the Tender coconut drink contained the lowest at 52.37 µg/ml. Additionally, the findings indicate that imported soft drinks generally contain higher caffeine levels than their domestic counterparts. These measurements are compared to the Recommended Daily Intake (RDI) for caffeine, providing insights into the potential consumption patterns of caffeine in soft drink products.
Contents
ABSTRACT
1.Introduction
1.1. General introduction
1.2. Statement of problem
1.3. Objective:
1.3.1. General objective
1.3.2. Specific objective
1.4. Limitations:
2. Literature Review
2.1. Soft drinks
2.2. Water
2.3. Sugar and sweetener
2.4. Acidity regulator and carbon dioxide
2.5. Flavoring and colorings
2.6. Preservatives
2.7. Caffeine
2.8. Health effects of Caffeine
2.9. Methods of Estimation of Caffeine
3. Materials and methods
3.1. Laboratory Setup
3.2. Research Design
3.3. Experimental Materials
3.3.1. Sample
3.3.2. Chemicals
3.3.3. Glassware
3.4. Instrument
3.5. Sampling
3.6. Methodology
3.6.1. Raw material collection
3.6.2. Determination of total dissolved solid (TDS) (AOAC, 2000)
3.6.3. Determination of pH (AOAC, 2000)
3.6.4. Estimation of Caffeine
3.6.5. Wavelength selection
3.6.6. Preparation of standards
3.6.7. Preparation of sample
3.6.8. Standard calibration curve
3.6.9. Calculation of Caffeine
4. RESULT AND DISCUSSION
4.1. pH
4.2. Total dissolved solute (TDS)
4.3. Standard calibration curve
4.4. Comparative study of soft drinks
5. Conclusion
6. References:
ABSTRACT
This study investigates the caffeine content in fourteen different local carbonated and energy drink brands, utilizing UV spectrophotometry for rapid and accurate caffeine quantification. The results reveal a significant variation in caffeine concentrations across the samples, with the energy drink XL(I) exhibiting the highest concentration at 520 µg/ml, while the Tender coconut drink contained the lowest at 52.37 µg/ml. Additionally, the findings indicate that imported soft drinks generally contain higher caffeine levels than their domestic counterparts. These measurements are compared to the Recommended Daily Intake (RDI) for caffeine, providing insights into the potential consumption patterns of caffeine in soft drink products.
1. Introduction
1.1. General introduction
Many plant species contain caffeine, an odorless, bitter chemical [1]. More than 63 plant species naturally contain it in their leaves, seeds, or fruits. Coffee (Coffea spp.), tea (Camellia sinensis), guarana (Paullinia cupana), maté (Ilex paraguariensis), kola nuts (Cola vera), and chocolate supply most caffeine. Guarana has the most caffeine (4–7%), followed by tea leaves (3.5%), maté tea leaves (0.89–1.73%), coffee beans (1.1–2.2%), cola nuts (1.5%), and cocoa beans (0.03%) [2]. Coffee, tea, soft drinks, cocoa, chocolate, medicines, and nutritional supplements include caffeine [3]. Cola and energy drinks include caffeine as a flavoring to increase their addictiveness [4].
Sugary, water-based beverages with balanced acidity are called soft drinks. The term "soft drink" means alcohol-free, unlike "hard drink". The word ‘drink’ is neutral but often indicates alcohol. Soft drinks can include a small amount of alcohol, but the concentration must be below 0.5% to be considered non-alcoholic. Soft drinks are refreshing beverages with 10-11% sugar, 0.3-0.4% acid (typically citric), flavoring, coloring, and chemical preservatives. Caffeine or fruit juice may be present. Vitamins, minerals, clouding agents, foaming agents, and plant extracts may be added [5].
Soft drink caffeine levels range from 10 to 50 mg per serving, whereas the FDA (2006) limits carbonated beverage caffeine to 6 mg per ounce. Thus, soft drink caffeine limits maybe 30 to 72 mg per 355 mL (12 oz) or 8.45 to 20.28 mg per 100 mL [4]. Caffeine stimulates the CNS, delaying sleep and improving study alertness [6]. It increases brain dopamine, which reduces depression [7]. However, excessive caffeine use can cause sleeplessness, anxiety, muscle twitching, migraines, pulmonary alkalosis, and palpitations [8].
1.2. Statement of problem
No nutritional advantage comes from caffeine. Caffeine is flavorless, making it hard to detect in our diets. Some medications include caffeine without our knowledge. Caffeine has several health effects. Caffeine overdose can cause general toxicity, cardiovascular issues, bone health and calcium balance changes, disorientation, hallucinations, emesis, adult behavioral problems, cancer risk, and male reproductive issues. Daily caffeine usage causes tolerance and addiction. Thus, sudden caffeine withdrawal may cause health problems. The issue is our inability to measure caffeine intake. Research is needed on caffeine’s health consequences.
1.3. Objective:
1.3.1. General objective
To measure the caffeine concentration in Coca-Cola, Sprite, Mountain Dew, Fanta, Pepsi, Red Bull, imported Coca-Cola, Pepsi, Mountain Dew, Red Bull, XL, Bullet, and lychee juice.
1.3.2. Specific objective
To measure caffeine in soft drinks. XL, Bullet, lychee juice, Coca-Cola, Sprite, Mountain Dew, Fanta, Pepsi, Red Bull, and foreign versions. To compare Indian soft drink caffeine levels to imported beverages. Soft drink pH and TDS measurement.
1.4. Limitations:
Local markets sell various soft drinks. However, my investigation will focus on a few soft drink samples. This is due to time and budget restrictions. Coffee, tea, energy drinks, and carbonated drinks contain caffeine, one of the most widely used psychoactive substances. Due to its stimulant properties, it is used in many beverages to boost alertness and attention. Modern society relies on caffeinated drinks to start their days and work all day. Soft drink caffeine concentrations fluctuate, making consistency difficult for customers. Understanding soft drink caffeine levels is important for various reasons. Caffeine can cause heart palpitations, anxiety, insomnia, and gastrointestinal issues. Thus, accurate caffeine content labeling helps customers choose beverages, especially those sensitive to caffeine or trying to limit their intake. Second, regulatory authorities, health organizations, and beverage makers use caffeine content data to make recommendations, guidelines, and product formulations.
The caffeine content in soft drinks depends on the beverage type, preparation technique, formulation, and portion size. Soft drinks can include caffeine from coffee beans, tea leaves, or synthetic caffeine. Different brewing processes and component ratios may affect caffeine levels among soft drink brands and flavors.
Research has measured caffeine levels in soft drinks and other liquids. Methodology, sample sizes, and testing techniques have led to conflicting results, preventing caffeine labeling and regulating guidelines. To keep up with changes and client preferences, the beverage industry needs constant research on new formulae and flavors.
Recent advances in analytical chemistry have made drinking caffeine concentrations more accurate. HPLC and GC-MS are common caffeine analysis methods that accurately and sensitively detect small amounts of the chemical. These analytical tools allow researchers to assess caffeine levels in soft drinks, revealing product variability and industry norms.
This study uses a systematic and rigorous approach to fix caffeine concentration gaps in soft drinks. We strive to provide a current and comprehensive caffeine content evaluation of popular soft drink brands and variants using modern analytical and sampling methods. We also aim to compare our results to current data and regulatory benchmarks to assess caffeine labeling precision and identify major anomalies that may require additional investigation or regulatory action.
This study affects the beverage industry’s public health, consumer awareness, and regulation. We help customers make informed decisions and promote producer accountability by clarifying soft drink caffeine levels and promoting clear labeling. Our findings may also help regulators improve regulations and monitor compliance to ensure the safety and integrity of caffeinated beverages. Assessing caffeine levels in various soft drinks is crucial for public health and consumer safety. This study uses rigorous research methods and powerful analytical tools to explore caffeine intake and its consequences on health and well-being.
Health Effects of Caffeine:
As a central nervous system stimulant, caffeine affects people differently based on age, weight, tolerance, and health. For most people, moderate caffeine usage is beneficial, but excessive use can be harmful. These may include anxiety, palpitations, insomnia, gastrointestinal trouble, and coffee poisoning. Researchers can better assess soft drink health risks and inform public health guidelines by properly characterizing caffeine amounts.
Consumer Education and Awareness:
Customers are carefully checking ingredients and nutritional information in this health-conscious era. Caffeine labeling helps people make informed choices that fit their diets and health goals. Knowing the caffeine content differences between soft drink brands and varieties helps customers make smarter beverage selections, promoting healthy consumption habits.
Compliance and Standardization:
Caffeine labeling standards and beverage industry compliance depend on regulatory bodies. However, caffeine measurement and labeling variations hinder regulatory oversight. Researchers assess caffeine levels in soft drinks and compare them to regulatory criteria to standardize labeling and enforce compliance. This boosts customer safety and trust in the beverage regulating system.
Industry and Formulation Trends:
The beverage industry is dynamic, with shifting client expectations and innovation. Understanding soft drink caffeine amounts reveals commercial tactics, formulation trends, and caffeine’s functional use. Tracking caffeine concentrations over time and across product lines can help researchers uncover formulation trends and assess their implications for consumer health and market dynamics.
International perspectives and culture:
Globally, caffeine is used, although its usage and cultural significance differ. Coffee is a staple in many countries, but soft drinks and energy drinks are becoming more popular, especially among young people. Comparing caffeine levels in soft drinks worldwide shows regional preferences, regulatory variances, and cultural attitudes toward caffeine. These results allow cross-cultural comparisons and inform global caffeine advocacy.
In conclusion, caffeine levels in soft drinks affect public health, consumer awareness, regulatory compliance, industry standards, and cultural impacts beyond quantitative measurement. Academics improve our understanding of caffeine’s function in modern beverage consumption and its social effects by studying these complicated features.
Different demographics and at-risk groups:
Pregnant women, toddlers, and those with certain medical conditions may be more sensitive to caffeine. Caffeine may affect fetal growth and pregnancy outcomes, so use caution. Due to their lower body weights and maturing brain systems, children and teens may be more caffeine-sensitive. Researchers can identify products that may be harmful to at-risk groups by examining caffeine levels in different soft drinks. This can inform public health and education activities.
Novel Drinking Trends: The beverage industry adjusts to changing consumer tastes and lifestyles. Innovative formulations and additives have resulted from the growing demand for healthier and more functional beverages. Understanding the caffeine levels in plant-based drinks, functional waters, and wellness beverages provides valuable information for altering beverage consumption patterns. Researchers can forecast consumer behavior and economic dynamics by studying caffeine trends in these growing categories, enabling supply chain players to act proactively.
Eco-friendly and ethical procurement:
Caffeine purchase and manufacture affect environmental sustainability and ethical sourcing in addition to nutritional and physiological considerations. Deforestation, soil degradation, and water shortages plague coffee and tea production, the main sources of natural caffeine. Concerns about fair trade, labor rights, and social justice in coffee and tea-producing regions emphasize ethical sourcing. Researchers may promote sustainable supply chains and openness about the beverage industry’s social and environmental impacts by evaluating soft drink caffeine concentration and component sourcing and manufacturing.
Psychoactive and behavioral effects:
The psychoactive properties of caffeine alter behavior, cognition, and physiology. While increasing alertness and focus, caffeine can also affect mood, attention, and decision-making. In academic, vocational, and recreational settings, understanding soft drink caffeine levels and their effects on cognition and behavior is crucial. By explaining how coffee affects behavior, researchers improve our understanding of human cognition and offer ways to improve performance and well-being.
Interactions with other ingredients:
Besides caffeine, soft drinks may contain sugars, artificial sweeteners, flavorings, and preservatives. Caffeine absorption, metabolism, and physiological effects may be affected by extra components. Caffeine, sugar, and artificial additives may worsen the health risks of excessive soft drink usage, such as obesity, metabolic issues, and tooth cavities. Researchers can better understand the health effects of soft drinks by studying their composition and caffeine interactions.
Understanding the impact of caffeine levels in soft drinks involves analyzing several complex factors. Researchers explore aspects such as caffeine intake, public health, industry practices, cultural influences, and environmental concerns to develop a framework that encourages responsible consumption of beverages. This approach not only aims to safeguard individual health but also contributes to societal and environmental well-being. By examining how caffeine affects the body, how industries formulate and market drinks, and how cultural perceptions vary, researchers work to create guidelines that promote safer consumption. Additionally, they focus on reducing the ecological footprint of beverage production, ensuring both human health and the planet’s sustainability are prioritized.
2. Literature Review
2.1. Soft drinks
Sugar-sweetened or "soft" drinks contain additional sugars or sweeteners, including high fructose corn syrup, sucrose, and fruit juice concentrates. This includes soda, pop, cola, tonic, fruit punch, lemonade (and other “ades”), sweetened powdered beverages, sports drinks, and energy drinks ("Sugary Drinks", 2019). Soft drinks are carbonated, non-alcoholic liquids. It includes juices, nectars, and sodas. Soft drinks are "soft" because they have less alcohol than hard drinks. Soft drinks cannot exceed 0.5% alcohol by volume [9]. Soft drinks can be carbonated or non-carbonated. Cola, lemon, and orange drinks are carbonated, whereas mango drinks are non-carbonated. Cola and non-cola soft drinks exist. Cola drinks, including Pepsi, Coca-Cola, Thums Up, Diet Coke, and Diet Pepsi, make up 61-62% of the soft drink business. Non-Cola products make up 36% and are available in Orange, Cloudy Lime, Clear Lime, and Mango flavors [10].
Young people drink soft drinks to quench their thirst in hot weather or because they think they help digestion, while athletes drink energy drinks to stay energized during intense physical activity and competition [11]. This study measures caffeine in numerous Indian energy beverages using UV-visible spectroscopy. Dichloromethane decreased absorbance to 274 nm. Sample 1 had the least caffeine, whereas Sample 3 contained the most. To avoid health risks from caffeine overuse, the authors recommend monitoring levels.
This research compares soft and energy drinks’ caffeine and pH. UV/Visible spectrophotometry found caffeine concentrations between 10.69 to 42.17 ppm. The lowest pH and highest caffeine content (42.17 ppm) indicated acidity in Brand 5. The authors warn that high caffeine and acidity levels in these beverages may pose health risks, suggesting that Brand 5 may need to rethink its economic viability.
This study uses dispersive liquid-liquid microextraction (DLLME) and gas chromatography-nitrogen phosphorus detection (GC-NPD) to quantify caffeine quickly. The process was used on teas, coffees, and eight beverages, including soft and energy drinks. High sensitivity and specificity made the approach fast and effective for caffeine analysis in complex matrices.
This study compares solvent extraction and activates carbon adsorption for carbonated and energy beverage caffeine measurement. The study found that extraction yielded 31.39 mg of caffeine per serving in Coca-Cola and 43.37 mg in Panther energy drink using spectrophotometric measurement. Adsorption produced somewhat higher values. Both methods work, however, the extraction technique matches product label values better.
This study assessed the caffeine content of 53 popular US nutritional supplements. The researchers found significant caffeine content variation amongst commodities, emphasizing the need for accurate labeling and client education. The results show that measuring caffeine intake from several sources is necessary to avoid health risks from excessive intake. This study developed and validated a fast HPLC method for caffeine quantification in Bangladeshi soft and energy drinks. The method was precise and accurate, with caffeine concentrations ranging from 20.11 mg/L to 121.56 mg/L among brands. The authors stress the need for caffeine regulation to protect consumers. This study examined solvent extraction and activated carbon adsorption for carbonated and energy beverage caffeine analysis. Through spectrophotometric measurement, extraction methods yielded 31.39 mg of caffeine per serving in Coca-Cola and 43.37 mg in Panther energy drink. Adsorption produced somewhat higher values. Both methods work; however, the extraction technique matches product label values better.
This research examined the caffeine levels in 53 widely used nutritional supplements in the U.S. The results revealed considerable variation in caffeine content among the products, highlighting the need for precise labeling and consumer education. The study emphasizes the importance of monitoring caffeine consumption from various sources to mitigate potential health risks linked to excessive intake.
To quantify caffeine accurately, the researchers employed Dispersive Liquid-Liquid Microextraction (DLLME) coupled with Gas Chromatography-Nitrogen Phosphorus Detection (GC-NPD). This method was particularly effective in analyzing caffeine levels in complex samples, such as teas, coffees, and a variety of beverages, including soft and energy drinks. It provided a rapid and reliable means of determining caffeine content.
Additionally, the research compared caffeine levels and pH values in soft and energy drinks. Using UV/Visible spectrophotometry, the team found caffeine concentrations ranging from 10.69 to 42.17 ppm. The highest caffeine content and lowest pH (42.17 ppm) were observed in Brand 5, which exhibited higher acidity. The authors caution that the combination of high caffeine and acidity in this product could present health risks, suggesting that Brand 5 may need to reconsider its market positioning and long-term economic feasibility.
This study reinforces the need for careful tracking of caffeine intake from different products to ensure consumer safety and minimize health risks associated with overconsumption. Table 2.1 shows the ingredients of soft drinks with their limit.
In Istanbul, Turkey, this study developed and validated an LC-MS/MS method for measuring caffeine levels in soft drinks such as iced tea, coffee, and energy drinks. The approach was precise and accurate, measuring caffeine concentrations from 37.96 mg/L in iced tea to 469.10 mg/L in coffee. Two of the 13 energy drinks tested exceeded the Turkish Food Codex’s caffeine limits, highlighting the need for tighter regulation.
This research introduced and validated a rapid High-Performance Liquid Chromatography (HPLC) technique to quantify caffeine in soft and energy drinks available in Bangladesh. The method was found to be both accurate and precise, with caffeine levels ranging from 20.11 mg/L to 121.56 mg/L across various brands. The study highlights the necessity of establishing caffeine regulations to ensure consumer safety.
This paper proposes a fast HPLC method for caffeine measurement in energy and cola drinks using UV diode array detection. The approach allowed high-throughput sample assessment with analytical times under 20 seconds. As caffeine amounts vary among the beverages examined, precise labeling and regulatory monitoring are needed to educate consumers on choices and ensure safety.
Young people should avoid coffee, according to new Healthy Eating Research guidelines sponsored by many medical organizations. Children aged 5 to 18 should consume mostly water and plain milk, restrict 100% juice and plant-based or flavored milk, and avoid sugar-sweetened, artificially sweetened, and caffeinated drinks. According to research, caffeine can cause sleep problems, high blood pressure, anxiety, and withdrawal symptoms in teens’ developing brains.
From 2022 to 2023, energy drink-related poison control calls increased by 24%, with 2,694 calls, mostly from unintended exposures in children aged 6–12. High caffeine and sugar in energy drinks can induce anxiety, irritability, sleeplessness, irregular heartbeat, and seizures in children, according to experts. The essay stresses the need for parents to educate their children about energy drink concerns and monitor their intake.
A fast Fourier Transform Infrared-Attenuated Total Reflectance (FTIR-ATR) spectroscopy technique was developed to measure caffeine in soft drinks without chemical solvents. Using PLS and PCR, the technique used the 2800-3000 cm⁻¹ portion of the FTIR spectrum for quantitative estimation. The PLS-1st derivative spectra showed an R² value over 0.97 and a SEP below 2.43. After recovery experiments and comparison with UV spectroscopic approaches, the process was shown to be effective for fast caffeine analysis.
This study examined caffeine in beverages using Solid-Phase Microextraction (SPME) and Gas Chromatography-Mass Spectrometry (GC/MS). The solvent-free approach required no pH changes and was acceptable for undergraduate labs. The approach accurately measured caffeine in coffee, tea, and cola, making it useful for education and analysis. This study automated caffeine quantification in soft drinks using Flow Injection Analysis (FIA) with amperometric detection. The method allowed direct sample insertion without treatment and 120 analyses per hour. Recovery rates ranged from 98% to 103%, with standard deviations from 2% to 5%. Comparing the strategy to the AOAC reference technique showed relative variations below 4%, demonstrating its accuracy and usefulness.
This work developed a fast caffeine measurement method using dispersive liquid-liquid microextraction (DLLME) and gas chromatography-nitrogen phosphorus detection (GC-NPD). The process was used on teas, coffees, and eight beverages, including soft and energy drinks. The caffeine analysis technique was sensitive and specific, making it fast and effective in complex matrices.
This study used reverse-phase HPLC with UV detection to measure caffeine in non-alcoholic energy drinks and teas. Nineteen beverages showed that energy drinks had the most caffeine. Many high-caffeine drinks were mislabeled, underlining the need for regulatory control to protect consumers.
Gupta and Mehta used HPLC with photodiode array (PDA) detection to measure caffeine levels in commercial fizzy beverages [12]. The research of 20 beverages found that cola-based drinks averaged 120 mg/L while energy drinks exceeded 500 mg/L. The study stressed the need for accurate caffeine measurement due to food safety regulations. The approach was sensitive enough to identify 0.05 mg/L, making it suitable for regular beverage analysis.
This study used graphene-modified electrodes to assess caffeine in soft drinks quickly. The sensor showed high selectivity and sensitivity, detecting 0.1 µM. Over 98% correlation was found between HPLC and the method. This method needed minimum sample preparation and provided real-time analysis, unlike chromatography. Electrochemical sensors can detect caffeine in beverages cost-effectively and sustainably, according to the study.
Oliveira and Santos measured caffeine in carbonated and energy drinks using NIR spectroscopy. Unlike conventional procedures, NIR spectroscopy allowed fast, non-destructive examination without chemical solvents. Chemometric models accurately measured caffeine concentrations between 50 and 600 mg/L [13]. The research showed that NIR spectroscopy can estimate accurately without harming samples, making it vital for beverage analysis and regulatory compliance.
A sustainable analytical chemistry paradigm was used to develop a spectrophotometric caffeine measurement method by Nakamura et al. [14]. The study replaced harmful organic solvents with ecologically friendly ones to reduce environmental effects and maintain analytical precision. The method showed a linear response from 10 to 500 mg/L caffeine, with a detection limit of 0.2 mg/L. Sustainable spectrophotometric approaches are ideal for soft drink caffeine monitoring since they match HPLC reference values.
Mina Kwon et al. predicted soft drink caffeine levels using machine learning and Raman spectroscopy [15]. Over 500 beverage samples were evaluated, and various machine-learning models were constructed to increase prediction accuracy. The best model had an R² value of 0.98 and an RMSE of 2.5 mg/L. The study showed the growing importance of artificial intelligence in food analysis, suggesting that data-driven caffeine measurement methods may improve efficiency.
Table 2.1 Ingredients of soft drinks with their limit [5].
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2.2. Water
Diet soft drinks might be 99% water, whereas regular ones are 90%. To reduce chlorine residual tastes, softened water is used. European Directive EC 98/1983, the US Environmental Protection Agency (EPA), and WHO recommendations to fulfill physical, chemical, and microbiological drinking water criteria.
2.3. Sugar and sweetener
Except for zero-calorie beverages, soft drinks have 1%–12% sugar (w/w). Sucrose, fructose, and glucose are common natural sweeteners. Also used as sweeteners include Acesulfame (E950), Sucralose (E955), Saccharin (E954), Thaumatin (E957), and Stevioside (E960).
2.4. Acidity regulator and carbon dioxide
Soft drinks have 1.5–5 g/L carbonation. It improves soft drink taste and preservation. Citric acid (E330) is used in soft drinks to boost antioxidants and add fragrance. EC regulation 1333/2008 governs food additives, including malic, succinic, and phosphoric acids, which provide acidity to drinks.
2.5. Flavoring and colorings
Soft drinks improve product aesthetics, correct processing-induced color changes, and preserve drink quality. There are three main color groups: natural, artificial, and caramel.
2.6. Preservatives
Soft drink preservatives improve microbiological stability. The chemical used depends on the preservatives and the soft drink’s chemical and physical properties. Sorbates and benzoates are often used in acidic drinks.
Hydrocolloids such as locust bean gum, pectin, and xanthan stabilize and thicken food and fruit juice drinks. Energy drinks include mostly taurine (average 3180 mg/L) and caffeine (360-630 mg/L). Energy drinks include vitamins B3, B6, and B12 [15].
Many beverages are consumed for their thirst-quenching or stimulating properties, not their nutritional value. Soft drinks are essential for hydration. Due to their osmolality, soft drinks absorb better than water, replace salt and energy, and quench thirst quickly. These drinks are among the top 10 providers of carbohydrates, vitamins, and minerals, according to research alongside energy. Soft drinks have three nutritionally significant regions. Beverages accounted for 20-24% of energy consumption. Some soft drinks provide customers with an immediate energy boost. Soluble sugars in soft drinks are easily digested. Second, isotonic drinks, which have an osmolality similar to physiological fluids, are nutritionally important. Fast electrolytes and water absorption make them vital for athletes and people who require fast hydration. Third, soft drinks have significantly evolved into low-calorie versions for those watching their calorie intake. Some firms claim their goods provide essential vitamins and minerals for children.
Soft drinks also harm health. Due to their high-calorie content, sugary drinks can cause weight gain. Soda dissolves tooth enamel. Researchers say fizzy beverages cause triple dental deterioration. Researchers have shown that fructose, especially from fizzy drinks, raises blood pressure. Due to soft drink usage, urine contains calcium and phosphoric acid. This causes osteoporosis [16].
2.7. Caffeine
German "kaffee" and French "café," both meaning coffee, are the origins of caffeine, technically known as 1,3,7-trimethylxanthine. In 2737 BC, Chinese Emperor Shen Nung boiled water and leaves from a nearby shrub to make the first pot of tea, which historians believe included caffeine. Caffeinated soft drinks like Dr. Pepper, Coca-Cola, and Pepsi-Cola appeared in the late 1800s. Caffeinated soft drinks were popular in the late 20th century due to their high caffeine content [3]. Friedrich Ferdinand Runge isolated pure caffeine in a lab in 1819 [17]. Figure 2.1. represented the structure of caffeine.
Three methyl groups on xanthine are swapped to become caffeine. Caffeine is C8H10N4O2. Aromaless, white, fluffy caffeine aggregates resemble dazzling needle-like powder. Its molecular weight is 194.19 g, melting point is 236 °C, sublimation point is 178°C at atmospheric pressure, pH of a 1% solution is 6.9, specific gravity is 1.2, volatility is 0.5%, vapor pressure is 760 mm Hg at 178 °C, and vapor density is 6.7 [18]. High boiling water solubility (66 g/100 mL) contrasts with moderate ambient water solubility (2 g/100 mL). It resembles cocaine and heroin. Caffeine activates the CNS via the same chemical mechanisms as other drugs (Mufakkar et al., 2014). The IUPAC name for caffeine is 1,3,7-Trimethylpurine-2,6-dione (Shar et al., 2017). Caffeine is an achiral molecule with two joined rings, a pyrimidinedione, and an imidazole. The nitrogen atoms are doubly linked to their neighboring amide carbon atoms, so all six atoms in the pyrimidinedione ring are ri.
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Fig. 2.1. Caffeine chemical structure Gerald et al. (2014).
One of the most often consumed active dietary components, caffeine, has been used for millennia. Coffee, tea, soft drinks, cocoa, chocolate, and medications contain caffeine.
Dietary supplements [3]. Mahoney et al. report that coffee (18%), soft drinks (18%), and tea (16%) are the main caffeine sources [19]. Worldwide, caffeine is the most consumed psychoactive chemical. Adults get three-quarters of their caffeine from coffee, whereas children get half from soft drinks and energy drinks [20]. A 355 mL (12 oz) soft drink can contain up to 71 mg of caffeine. Lack of government oversight has led to extensive advertising of energy drinks with high caffeine levels, especially for psychotropic, performance-enhancing, and stimulant effects on young men. Caffeine intoxication from energy drinks is rising, suggesting caffeine dependency. Lack of pharmacological tolerance makes non-caffeinated children and adolescents more susceptible to caffeine intoxication. Numerous research suggests energy drinks may lead to other drug dependence. Due to its harmful effects, some governments have set maximum caffeine restrictions for soft drinks [21].
Table: 2.2 Caffeine regulation in numerous countries [21].
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2.8. Health Effects of Caffeine
A lot of caffeine is consumed. Caffeine dissolves in water and lipids, crosses the blood-brain barrier, and is found in all body fluids, including saliva and cerebrospinal fluid. The umbilical cord and breast milk will show caffeine from perinatal moms. Thus, the fetus and breastfed infants will have it. The small intestine absorbs caffeine entirely in less than an hour and diffuses it into other tissues. Small intestine absorption appears to be unaffected by sex, genetics, environment, or other factors, although further study is needed. Saliva caffeine levels peak 45 minutes after consumption and serum 2 hours later. People’s caffeine half-life is 3–7 hours. Neonates’ undeveloped kidneys and livers extend the half-life to 65–130 hours. Caffeine’s effects depend on tissue duration; therefore, peak concentrations matter. Age and complex genetic and environmental combinations affect the consequences [22]. The liver breaks down caffeine into paraxanthine (84%), theobromine (12%), and theophylline (4%). Paraxanthine increases plasma-free fatty acids. Urine volume rises with Theo bromine. Theophylline relaxes smooth bronchial muscles and treats asthma. Fig 2.2 represents the caffeine and metabolites.
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Fig: 2.2 Caffeine and metabolites (Tautua et al., 2014).
Students use caffeine during late-night study sessions to reduce fatigue and boost attentiveness. Sprinting, endurance, and teamwork can improve.
In anemic individuals, caffeine suppresses erythrocyte apoptosis, preventing a drop in count. Caffeine aids long-term memory [23]. According to a Duke University study by Sinha et al., caffeine may reduce fatty liver in non-alcoholic fatty liver disease patients and liver fibrosis in hepatitis C patients [24]. At 100 mg per day, caffeine may be prevented [25]. German research found that weight-loss participants who drank 2-4 cups of caffeinated coffee daily were more likely to sustain their weight reduction than those who did not [26]. Caffeine also stimulates brown adipose tissue, increasing caloric expenditure and metabolism. It also keeps drivers aware during sleep deprivation [27]. Caffeinated coffee reduces melanoma risk. Coffee drinkers are less likely to commit suicide [28]. Coffee reduces the incidence of Parkinson’s disease, especially for genetically susceptible individuals [29]. A large research study of 217,883 people found that coffee from any source reduces kidney stone formation. Caffeine increases urine dilution [30]. Bigotte et al. found in 2018 that caffeine reduces renal disease mortality [31].
To avoid headaches, tiredness, anxiety, and nausea, caffeine-sensitive people should restrict their daily intake to 400 mg [3]. Excessive caffeine usage during pregnancy can cause miscarriage, difficult labor, and low-birth-weight babies. Caffeine-containing foods exceeding 150 mg/L must be labeled [8]. Nowadays, regular caffeine usage beyond 500–600mg is considered ‘abuse’ due to its health risks. Caffeine addiction can cause ‘caffeinism,’ which has several detrimental repercussions. Caffeine over 400mg/day may increase the risk of detrusor instability in women. Moderate caffeine usage (200–400mg/day) may increase detrusor instability in women with bladder symptoms [32].
A few examples of excessive caffeine usage have been recorded. The estimated acute lethal dose for adults is 10 g. A patient survived despite consuming 24g of caffeine, but 6.5g was killed. Adenosine receptor antagonism is caffeine’s main action. Caffeine is quickly and practically completely absorbed from the gut into the bloodstream. Peak blood caffeine levels occur 1–1.5 hours after intake. Caffeine is quickly absorbed and distributed. It crosses the blood-brain barrier, enters amniotic fluid and the fetus, and is in breast milk. Semen contains caffeine. Only 1–5% of caffeine is eliminated unchanged in urine. The urine of 8–9-month-olds contains 85% of the caffeine they eat unaltered [33].
Mufakkar et al. found that Pepsi had the highest caffeine content among eight Pakistani soft drink brands, with 255.27 µg/mL, followed by Coke (211.69 µg/mL), Fanta (357 µg/mL), Mountain Dew (119.42 µg/mL), and Sprite (30.99 µg/mL) [34].
2.9. Methods of Estimation of Caffeine
There are several ways to measure methylxanthine in food and beverages. UV-visible spectrophotometry, potentiometry, HPLC, ion chromatography, HPTLC, capillary electrophoresis, micellar capillary electrophoresis, gas chromatography, and solid-phase microextraction gas chromatography are the methods [35].
Pharmaceutical analysis often uses UV-visible spectrophotometry. It measures how much UV or visible light a solution absorbs. UV-visible spectrophotometers measure the ratio or function of the intensity of two UV-visible light beams.
I’m ready to utilize UV spectrophotometers at the college lab because the equipment is easy to operate.
What Caffeine Does to Cognitive Performance:
Numerous studies have shown that caffeine boosts alertness, concentration, and cognitive ability. Nehlig et al. found that moderate coffee intake improves response speed, alertness, and memory in a meta-analysis of over 40 randomized controlled trials [36]. Caffeine responses vary by age, genetics, and drinking habits. More research is needed to understand caffeine’s cognitive-enhancing properties and its potential usefulness in neuroprotection and enhancement.
Soft Drink Consumer Perceptions and Preferences:
Understanding consumer perception and preferences is essential for soft drink product development and marketing. Volkner and Hofmann examined soft drink customer preferences for flavor, packaging, brand impression, and healthiness [37]. The findings showed that flavor and brand impression influence customer choices and that consumers want healthier, more natural drinks. The study showed how sensory marketing affects consumer perceptions and purchases. This study can inform product innovation and positioning to meet client wants.
Beverage Caffeine Labeling Regulations:
To improve transparency and customer safety, global regulatory agencies require caffeine content labeling in drinks. Temple et al. examined regulatory measures in different countries and organizations, highlighting variations in labeling, caffeine limits, and transparency [22]. Some jurisdictions enforce caffeine labeling on beverage packaging, while others demand voluntary disclosure or limit caffeine in particular drinks. These guidelines are difficult to reconcile due to risk evaluations, cultural standards, and industry advocacy. Standardizing caffeine labeling helps customers make educated decisions.
Environmental Impact of Soft Drink Packaging:
Due to the knowledge of plastic waste and resource depletion, soft drink packaging sustainability is a major problem. Parker et al. examined the life cycle environmental impacts of beverage packaging materials such as plastic bottles, aluminum cans, and glass containers [38]. Energy use, greenhouse gas emissions, and waste creation varied widely among packaging methods, with plastic bottles having the most environmental impact. Material substitution, recycling, and eco-design can reduce soft drink packaging’s environmental impact and promote a circular economy.
Adolescent Caffeine Trends:
Teens have unique caffeine intake patterns and health impacts. Bernstein et al. studied what influences teens' caffeine intake in a longitudinal study [39]. Caffeine consumption increases during adolescence, mostly from coffee, soft drinks, and energy drinks. Teen caffeine use was influenced by socioeconomic status, peer influence, and advertisement. Appropriate treatments are needed to promote safe beverage choices and decrease caffeine-related harm in adolescents.
Social Media’s Effect on Adolescent Mental Health:
Numerous research studies have examined how social media use affects teens’ mental health. Twenge and Campbell investigated longitudinal and cross-sectional data and found a consistent link between substantial social media use and teen sadness, anxiety, and loneliness [40]. Cyberbullying, social comparison, and sleep changes may mediate this relationship, although its direction and mechanisms remain unclear. The complex link between social media use and adolescent mental health needs further investigation.
Working remotely affects employee productivity and well-being:
Remote work models are widely used, therefore, their effects on employee productivity and well-being are being studied. Tammy D. Allen et al. [41] examined the impact of remote labor on numerous outcomes using empirical studies and meta-analyses. Some studies show benefits to job satisfaction, work-life balance, and autonomy, while others show social isolation, blurred work-life boundaries, and decreased teamwork. Flexible scheduling, virtual team-building, and organizational support systems may maximize remote work and minimize its drawbacks.
How AI Advances Healthcare:
AI technology has the potential to transform healthcare and improve patient outcomes. Topol examined AI in diagnostic imaging, medication development, predictive analytics, and tailored medicine [42]. AI algorithms have improved disease detection, risk classification, and therapy optimization, thus improving clinical decision-making. However, data privacy, algorithmic bias, and regulatory monitoring must be addressed to ensure ethical AI use in healthcare.
Climate Change Impacts Global Food Security:
Climate change threatens global food security by reducing agricultural production, food supply, and nutrition. Wheeler and von Braun conducted a meta-analysis of studies on how climate variability and extreme weather affect agricultural productivity and food costs [43]. The findings showed that low-income countries and smallholder farmers are more vulnerable to climate change. Crop diversity, irrigation infrastructure, and weather-indexed insurance programs are vital for food system resilience and climate change mitigation.
Artificial Intelligence and Robotics Ethics:
AI and robotics ethics have been scrutinized in academia and industry. Floridi et al. examined the ethical frameworks and principles that guide AI and robotic system invention, deployment, and regulation [44]. Algorithmic discrimination, privacy violations, job displacement, and safety-critical autonomous decision-making are major ethical challenges. Multidisciplinary collaboration, stakeholder participation, and regulatory frameworks to preserve ethical standards and societal norms are needed to address these issues. Trust and AI and robot benefits depend on balancing scientific progress and ethical responsibility.
Physical Activity and Mental Health:
Schuch et al. meta-analyzed over 49 randomized controlled trials on exercise and mental health [45]. After constant exercise, depression and anxiety decreased significantly. Exercise also improved self-esteem, cognition, and well-being. These benefits result from endorphin release, inflammation decrease, and neurogenesis improvement. Exercise is a non-pharmacological way to improve mental health, and this study recommends including it in therapy for many psychiatric diseases.
Social Support and Chronic Disease Management:
A comprehensive analysis by Uchino compiled longitudinal studies on social support and chronic illness management [46]. Chronic diseases, including diabetes, cardiovascular disease, and cancer, were reviewed. Multiple studies found that social support enhanced treatment adherence, disease management, and health outcomes. Additionally, social support mitigated the negative effects of stress on physiological functioning and immunological regulation. These findings demonstrate the need to incorporate social support into chronic illness care.
Nutrition’s Effect on Older Adult Cognitive Function:
A literature review by Smith and Blumenthal examined nutrition and cognitive performance in elderly adults [47]. The study included observational and clinical data on how nutrition, macronutrients, and nutrients affect cognitive decline and dementia risk. The findings showed that a Mediterranean diet rich in fruits, vegetables, whole grains, and healthy fats can prevent cognitive decline. Omega-3 fatty acids, antioxidants, and vitamin B12 improve cognition and reduce neurodegenerative disease risk. Dietary interventions are important for cognitive performance in elderly populations.
Stress Reduction from Meditation:
A meta-analysis by Goyal et al. examined 47 randomized controlled trials to determine if meditation reduces stress [48]. The evaluation covered mindfulness, transcendental, and loving-kindness meditation. Meditation therapy significantly reduces stress, anxiety, and depression. Meditation lowered cortisol and increased heart rate variability. According to the study, medical education improves stress management and psychological well-being.
Urban Green Spaces and Public Health:
Gascon et al. examined the health effects of parks, gardens, and natural landscapes in cities. The analysis combined epidemiological evidence on green environments and varied health outcomes [49]. Closeness to green spaces lowered cardiovascular disease, obesity, mental illnesses, and premature mortality. Engaging with nature improves mood, cognition, and immunity. These findings highlight the importance of urban planning initiatives that prioritize green space to improve public health and well-being.
3. Materials and methods
3.1. Laboratory Setup
The entire project was done in Campus labs.
3.2. Research Design
Laboratory experiments are used in the investigation.
3.3. Experimental Materials
3.3.1. Sample
Soft drink
3.3.2. Chemicals
Anhydrous caffeine powder (98-101.5%) with pH 4 and pH 7 buffer tablets and distilled water.
3.3.3. Glassware
Volumetric flask, beaker, pipette, funnel, measuring cylinder, watch glass, and glass rod.
3.4. Instrument
Labtronics LT-291 UV-Vis Spectrophotometer with pH meter, Digital balance, and Conductometer.
3.5. Sampling
The department store provided a soft drink sample.
Table 3. 1 Soft drink sample specifications.
Illustrations are not included in the reading sample
3.6. Methodology
3.6.1. Raw material collection
The Departmental Store has fourteen soft drinks. The 100-ml samples were selected using basic random sampling.
Illustrations are not included in the reading sample
Fig. 3.1 The Soft drink samples [Author’s work]
3.6.2. Determination of total dissolved solid (TDS) (AOAC, 2000)
Conductivity meters measured total dissolved solids. The measuring head was submerged in 10 ml of soft drink in a beaker.
3.6.3. Determination of pH (AOAC, 2000)
Soft drink acidity was measured by pH. A pH meter calibrated with pH 4 and 7 buffer solutions tested the pH of 5 ml of soft drink in a beaker.
3.6.4. Estimation of Caffeine
Caffeine calculation follows Mufakkar (2014) with some adjustments [34].
3.6.5. Wavelength selection
The wavelength was obtained by scanning 120–400 nm. According to Mufakkar (2014), caffeine was measured at 272 nm [34].
3.6.6. Preparation of standards
To make a 400 µg/ml caffeine stock solution, dissolve 0.04 grams of caffeine in 100 ml of distilled water in a 100 ml volumetric flask. The stock solution was stored at +4ºC in a dark place. Standard solutions of 0.01, 1, 5, 10, 15, and 20 µg/ml were created by diluting the stock solution.
3.6.7. Preparation of sample
One milliliter of samples will be placed in a 100-milliliter volumetric flask, and distilled water will be added to the calibration mark. Sample absorbance will be measured with a UV spectrophotometer. Calibration curves determine absorbance-based concentration.
3.6.8. Standard calibration curve
A UV-Vis Spectrophotometer (Labtronics, Model: LTetric-291) measured caffeine working standard solution absorbance at 271 nm. A graph of absorbance vs. concentration yielded the standard calibration curve.
3.6.9. Calculation of Caffeine
A standard calibration curve was used to calculate caffeine contents from the 272 nm absorbance of different soft drink samples.
4. RESULT AND DISCUSSION
This research estimated soft drink caffeine levels. This study measured caffeine in 14 soft drink samples. Along with caffeine, pH and total dissolved solids were assessed in soft drink samples. Laboratory analysis yielded the following results.
Table 4.1 pH and TDS value of soft drinks.
Illustrations are not included in the reading sample
4.1. pH
The pH of soft drinks ranged from 2.42 to 5.49. Imported Coke was the most acidic among the samples. Tender coconut is the least acidic soft drink. Low pH may be caused by carbon dioxide and phosphoric acid. These beverages include preservatives including malic and tartaric acids [17]. These acids inhibit the growth of bacteria, mold, and fungi that taint drinks.
4.2. Total dissolved solute (TDS)
The aggregate concentration of solubilized particles in water. BIS sets a maximum intended total dissolved solids (TDS) level of 0.5 ppt and a maximum acceptable level of 2 ppt in the absence of superior water sources [16].
The highest and lowest TDS values were found in Indian Tender (4.930 ppt) and Bottlers India Dew (0.205 ppt). Significant dissolved material concentrations seldom cause health danger. However, low TDS levels make the water taste bland, which many people dislike.
4.3. Standard calibration curve
The picture shows the UV-Vis spectrophotometry caffeine calibration curve.
Illustrations are not included in the reading sample
Fig: 4.1 Calibration curve [Author’s work]
The standard curve showed a linear relationship between caffeine solution content and absorbance. The linear equation y = 0.047x - 0.009 describes absorbance and concentration. Applying a linear trend line to the data set yields a high R² value of 0.999, indicating a strong fit.
Table: 4.2 Absorbance and corresponding concentration in µg/ml.
Illustrations are not included in the reading sample
The caffeine concentration in soft drinks varied from 52.37 to 520.88 µg/ml. Caffeine concentrations measured: Coke (I): 298.66 µg/ml, Red Bull (I): 492.56, Pepsi (I): 318.47, Red Bull (N): 318.46, Bullet (I): 488.32, XL (I): 520.88, Litchi juice: 87.05, Coke: 285.21. The highest caffeine content was found in XL (520.88 µg/ml), whereas the lowest was found in another sample. The concentration of caffeine in litchi juice and Sprite was 87.05 µg/ml. This matches [34].
Table 4.3 RDI and total daily consumption volume.
Illustrations are not included in the reading sample
Illustrations are not included in the reading sample
Fig: 4.2 Standard caffeine solutions [Author’s work]
The table above shows how many soft drinks people can drink to avoid exceeding caffeine consumption. Bullet (I), XL (I), and Red Bull (I) contain high caffeine levels, therefore avoid drinking more than 819.13, 767.93, and 812.08 ml of each. Coke (I), Pepsi (I), Coke (N), and Pepsi (N) should be consumed in 1339.32ml, 1256.01ml, 1402.48ml, and 1371.84ml portions.
4.4. Comparative study of soft drinks
Illustrations are not included in the reading sample
Illustrations are not included in the reading sample
Fig: 4.3 Comparative Soft Drink Caffeine Concentration [Author’s work]
Soft drinks from India and other countries are compared. The imported Coke has 298.66 µg/ml of caffeine, whereas the domestically made one has 285.21. Also, imported Pepsi includes the caffeine content in one beverage is 318.47 µg/ml, whereas Indian Pepsi contains 291.58 µg/ml. The caffeine content in imported Dew is 263.27 µg/ml, but Indian Dew has just 220.1 µg/ml. This difference may be due to processing facilities. Red Bull made in India has 381.46 µg/ml of caffeine, whereas imported Red Bull has 492 µg/ml. The investigation found that all international samples had more caffeine than Indian ones.
5. Conclusion
- XL has the highest caffeine level at 520.88 µg/ml, while Litchi juice and Sprite have the lowest at 87.05 µg/ml.
- Imported soft drinks have more caffeine than Indian ones.
- Multiple soft drinks can be caffeine-estimated.
- Caffeine, phosphoric acid, and other soft drink ingredients may be measured.
- Soft drinks can be tested for heavy metals.
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