The Caffeine Content of Energy Drinks in accordance with the Information on the Package Label

. Six diferent brands of energy drinks (EDs) were analyzed to determine the concentration of cafeine in accordance with the information on the package label. Approximately 28.1 milligrams per 100 milliliters (mg/100ml) of cafeine was present in the Mo energy drink brand, which was nearly 1.5 times the cafeine concentration listed on the product label. Te energy drink with the highest cafeine content was Dragon, which contained 30.1mg/100ml, or a total of 150.5mg per 500ml bottle. Manufacturers of energy drinks (EDs) should accurately list the amount of cafeine and other ingredients on the product label so that consumers know how much of each ingredient they are taking. Taking too much cafeine can be bad for your health.


Introduction
Energy drinks (EDs) are nonalcoholic, sugary beverages that contain variable amounts of cafeine, taurine (amino acid), glucuronolactone, herbal extracts, minerals, and vitamins in order to increase physical and mental endurance [1].Te majority of people who consume EDs are engaged in strenuous physical activity, as well as those who prefer to remain alert for longer durations, such as athletes, drivers, and students.Because EDs are marketed for their perceived or actual benefts as stimulants, performance enhancers, and energy boosters, this is the case [2].EDs were frst introduced in Asia and Europe around 1960 [1].In 1987, when the most well-known brand, Red Bull, was introduced, its consumption became very popular [3].
Due to the widespread and long-term consumption of beverages containing cafeine naturally, such as cofee and tea, the regulation of beverages containing added cafeine has been complicated [4].Nevertheless, a number of nations have enacted regulations governing the labeling, distribution, and sale of energy drinks containing signifcant amounts of cafeine [4].Te European Food Safety Authority mandates that energy drinks with a cafeine content greater than 150 parts per million (ppm) or 150 mg per liter must be labeled as having "high cafeine content" and the exact amount must be specifed [5].Australia has banned energy drinks with more than 320 ppm of cafeine and wants to put them in the same category as drugs.Countries such as Denmark, Uruguay, and Turkey have banned energy drinks entirely, and Sweden has prohibited the sale of energy drinks to minors [5].Te maximum recommended daily cafeine intake varies between 2.5 mg/kg/day and 6 mg/kg/day for children, 100 mg/day for adolescents, and 400 mg/day for adults [6].Tolerance to cafeine varies between individuals, with the majority of people developing toxic symptoms at doses of 200 mg [7].
A study conducted by CSE [4] to determine the cafeine content of energy drinks revealed deviations from the amount of cafeine listed on the label, as shown in (Table 1).
Joo et al.found that the health risks associated with ED consumption are primarily attributable to their cafeine content.An overdose of cafeine can cause palpitations, hypertension, dieresis, stimulation of the central nervous system, nausea, vomiting, marked hypocalcemia, metabolic acidosis, convulsions, and, in rare cases, death [9].Tere is also an increased risk of arterial hypertension and Type 2 diabetes in adults due to the fact that high cafeine consumption reduces insulin sensitivity [10].Cafeine consumption increases the risk of late miscarriages, smallfor-gestational-age infants, and stillbirths among pregnant women.Frequent cafeine consumption may impair cognition in general and long-term memory and learning in particular [11].Cafeine at a dose of 400 mg increases anxiety, particularly when combined with a stressful task.Some studies suggest that EDs can trigger the onset of the frst seizure and contribute to stroke [12].
Cafeine overconsumption is linked to a variety of additional health issues.Cafeine overdose, for instance, has been reported as a potential cause of rhabdomyolysis [13].Due to their diuretic efect, EDs can cause hypokalemia as well as high creatinine kinase levels and renal impairment [14].Individuals who do not regularly consume large quantities of cafeine may experience an increase in diuresis after consuming EDs.Terefore, dinks have an overall dehydrating efect [15].In addition, Hasselkvist et al. [16] found a signifcant relationship between EDs and dental erosion.Te low pH of energy drinks is largely responsible for their high demineralization potential [17].According to Vos and Lavine [18]; increased consumption of sugar-sweetened beverages, including ED, is associated with overweight and obesity, as well as an increased risk of diabetes mellitus and cardiometabolic diseases because beta cells are unable to secrete enough insulin to maintain normal blood glucose levels when exposed to excesses of simple sugars for extended periods of time.
Cafeine, the primary component of the drinks, stimulates sympathetic nerves, which can cause adverse health efects, such as sudden death .
Despite an increase in the number of industries in Tanzania producing various brands of EDs and the presence of imported products, there are no published studies in Tanzania assessing the concentration of cafeine in ED products in accordance with the information on the package label.Te study's overarching objective is to determine the amount of cafeine used in energy drink products in accordance with the labeling information.

Estimation of Sample Size for Energy Drinks.
Purposeful sampling was used to collect twelve energy drink samples, two from each of six brands sold in Tanzanian grocery stores and convenience stores.Two grocery stores were visited and the shopkeepers were asked to select six samples of EDs beverages that were most likely preferred by customers.Te other three must consist of brands produced in Tanzania, and the other three must consist of brands imported from outside the country; this was done in both grocery stores.

Data Collection.
Te samples were randomly obtained by visiting two grocery stores and asking the shopkeepers to select six samples of EDs beverages that were most likely preferred by customers.Tree of the samples had to be Tanzanian brands, and the other three had to be foreign brands; this was done in both grocery stores.Te samples were placed in a cool box with icepacks and transported within one to two hours to the Food Science and Technology Laboratory at Sokoine University of Agriculture (approximately 2.5 km from the sample collection sites), where they were stored at a temperature of 1.6 °C until analysis.

Procedures for Cafeine Extraction and Sample
Measurements.Each working standard and sample solution yielded 1.0 mL of homogenized beverage.Tis sample was placed in a separating funnel, 1 ml of 20% (w/v) methanol and 5 ml of chloroform were added, and the mixture was shaken for fve minutes.Te absorbance of the lower (organic) layer containing cafeine was measured at 274 nm using a UV Vis spectrophotometer (X-ma 3000, England).Te extraction method was performed twice for each sample (Table 2).
A 1000 ppm standard stock solution was prepared by dissolving 0.1 g of pure cafeine in 100 ml of chloroform.We prepared serial dilutions of 1 to 25 ppm, measured absorbance, determined a linear regression equation, and used Microsoft Excel to tabulate measurement data and conduct a linear regression analysis.Tis allowed for the calculation of a concentration factor, which was then used to quantify the cafeine.Te fnal cafeine content of the tested beverage was then calculated using the concentration of the extracted sample solution and the following equation (Figure 1): Caffeine content in milligrams � Conc(ppm) ×(total sample volume(ml)2).

Analyses of the Actual Amounts of Cafeine Found in
Energy Drink Ingredients.Te six diferent brands of ED beverages in duplicates (a total of 12) consisted of three brands that were produced in Tanzania (Azam energy drink, Mo energy drink, and Power energy drink) and three that were imported from abroad (Kungfu energy drink, Dragon energy drink, and Monster energy drink).Of the six samples tested, the sample of Dragon energy drink had the highest cafeine content at 30.1 mg/100 ml, resulting in a total of 150.5 mg of cafeine per bottle (Table 3).

Advances in Public Health
Cafeine content samples (BEVERAGES) analyzed in duplicate (S1 and S2) during a 2018-2019 study of energy drink consumption rate, knowledge, and perceived side efects among long-distance car drivers in Morogoro municipality, Morogoro, Tanzania (Table 3).
Monster energy drink had the highest cafeine content per serving (per packaged bottle), followed by Dragon energy drink and Kungfu energy drink, followed by Power energy drink and Azam energy drink, with Mo energy drink having the least amount of cafeine indicated on its package label (Tables 4 and 5).

Discussion
Since cafeine is the main ingredient in energy drinks, cafeine overdoses are the leading cause of adverse health efects [19].Due to its higher cafeine content, some nations, including Turkey, have banned its consumption [20].Referring to the current FDA regulation, "dietary ingredients that are specifcally added to products must contain 100 percent of the volume or weight declared on the label (with the exception of an analytical method-attributable deviation), and whereas naturally occurring dietary ingredients must be present at 80 percent of the declared value" [21,22].In our study, we compared the cafeine concentrations of six EDs samples (Azam, Dragon, Kungfu, Mo, Monster, and Power) with the cafeine content listed on the product's label.Five of the examined EDs contained less cafeine than indicated on the product label.However, the Mo energy drink contained approximately 1.60 times the amount of cafeine as indicated on the product label.Tis study's fndings are comparable in some ways to those reported by Consumer Reports [23]; which examined the cafeine content of 16 energy drinks and found that 10 of the samples had values within 20% of the actual numbers listed on the label, while 5 of the product samples had cafeine amounts greater than 20% above what was indicated on the label, and 1 product had cafeine amounts 70% below what was indicated on the label.Similar research [24] revealed that the majority (89%) of the EDs samples they analyzed contained cafeine within 20% of the labeled amounts.A study [20] found that 100 ml of EDs contains between 80 and 242 mg of cafeine, which is equivalent to consuming eight cups of strong cofee per day, thereby increasing the risk of toxicity and disrupting the overall health structure.Advances in Public Health

Conclusion
Manufacturers should accurately label the cafeine concentration on their energy drink products, and consumers are advised to use the recommended amount of cafeine in order to avoid adverse health efects that may result from excessive consumption.Additionally, consumers should avoid energy drink products that do not adhere to the cafeine concentration indicated on the package label.
5.1.Recommendations.Te government and responsible agencies should take the necessary steps to ensure that all energy drinks sold in the United States contain the recommended amount of cafeine, which will be specifed on the packaging label, in order to reduce the risk of cafeinerelated side efects.

Table 2 :
Absorption of fnal solutions at 274 nm.

Table 1 :
Te cafeine content of popular energy drinks.

Table 3 :
Results on cafeine contents samples (beverages) analyzed in duplicate (S1 and S2) during a study to assess energy drink consumption rate, knowledge, and perceived side efects among long-distance car drivers in Morogoro municipality, Morogoro, Tanzania, 2018-2019.

Table 4 :
Cafeine concentration per serving for various energy drinks according to their package labels.

Table 5 :
Laboratory results for the cafeine content of the beverage samples and the published cafeine content according to the manufacturer's package label Morogoro municipality, Morogoro, Tanzania, 2018-2019.