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Determining the caffeine content in different tea brands

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Investigating the caffeine content (mg) in different brands of black tea (Tetley

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Black Tea, American Breakfast Black Tea, Filiz Black Tea, & Red Label Tea)

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using dichloromethane in a separating funnel
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Chemistry SL Internal Assessment

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Page count: 12
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Candidate Number: kpm700 (003877-0008)

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Table of Contents
1. Introduction …………………………………………………………………………………. 3
2. Background ………………………………………………………………………………….. 3
2.1 Hypothesis ………………………………………………………………………………… 5
3. Variables …………………………………………………………………………………....... 5
3.1 Independent Variables …………………………………………………………………….. 5
3.2 Dependent Variables ……………………………………………………………………… 5
3.3 Controlled Variables ………………………………………………………………………. 6
4. Apparatus ……………………………………………………………………………………. 6

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4.1 Photo of setup ……………………………………………………………………………... 7

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4.2 Safety Precautions …………………………………………………………………………. 7

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5. Procedure …………………………………………………………………………………….. 7
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6. Quantitative Data …………………………………………………………………………… 8
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6.1 Raw Data ………………………………………………………………………………….. 8

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6.2 Processed Data …………………………………………………………………………….. 9

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6.2.1 Sample Calculations …………………………………………………………………… 9

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7. Qualitative Data ……………………………………………………………………………. 12

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8. Conclusion ………………………………………………………………………………...... 12
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9. Evaluation …………………………………………………………………………………... 12
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9.1 Strengths ……………………………………………………………................................. 12

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9.2 Weaknesses and Limitations ……………………………………………………………... 13

9.3 Improving the experiment ………………………………………………………………... 13
9.4 Extension …………………………………………………………………………………. 14
10. References …………………………………………………………………………………. 15

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1. Introduction
After water, tea is the beverage that is consumed the second most frequently all across the world.
Camellia sinensis, the plant from which tea is derived, is the progenitor of all the different types
of tea. Black tea is one of the many varieties of tea that can be produced as a result of the plant's
harvest and subsequent processing in one of several distinct ways. It is common practice to
combine black tea with other plants, such as English Breakfast and Earl Grey, in order to give
black tea a more complex flavor and scent. Additionally, it is well-known for the amount of
caffeine it contains, which is far higher than that of its competitors, such as green or white tea.
However, compared to coffee, the amount of caffeine that is found in black tea is far lower
(TIMESOFINDIA.COM, 2020).
My interest was piqued when I came across this article, and because I am a busy person and a
student, it is often difficult for me to stay focused. However, I always find solace in a cup of tea.

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Through this experiment, I will be able to determine which types of tea are best for me based on
the amount of caffeine that is contained in each type of tea, which will enable me to remain

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focused. With that being said, the research question “Investigating the caffeine content (mg) in

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different brands of black tea (Tetley Black Tea, American Breakfast Black Tea, Filiz Black

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Tea, & Red Label Tea) using dichloromethane in a separating funnel” with the help of the
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solvent, dichloromethane, which is produced by chlorinating methane through a separating
funnel.
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2. Background
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Dichloromethane
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Dichloromethane, also known as methylene chloride is a chlorinated hydrocarbon that is a

transparent, colorless liquid that is volatile and nonflammable. It has a sweet, pleasant smell, and
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when it is heated to the point where it decomposes, it emits very poisonous vapors of phosgene.
Methylene chloride is most commonly employed as a solvent in paint removers; however, it is
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also utilized in the formulation of aerosols, as a solvent in the production of pharmaceuticals, as

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a degreasing agent, in the production of electronics, and as a blowing agent for ethane foam.
When inhaled, this chemical causes irritation to the respiratory tract, specifically the nose and
throat, as well as the central nervous system. Methylene chloride has the potential to cause
genetic mutations and has a high probability of being carcinogenic to humans (PubChem,
Dichloromethane, 2019).

Figure 1: Structural Formula (Dichloromethane - American Chemical Society, 2019)

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Caffeine
Caffeine (C8H10N4O2) is a purine alkaloid that belongs to the xanthine class of compounds. It is a
white, crystalline substance that is highly water-soluble and has a bitter taste (Caffeine Structure,
2021).
Caffeine is synthesized from xanthine, a compound that is produced by the breakdown of
adenine, a nucleotide found in DNA and RNA. The synthesis of caffeine occurs in the leaves,
seeds, and fruits of many plants, and it is thought to serve as a natural insecticide and protectant
against herbivores. The methylation of theobromine that had been isolated from cocoa yielded
the majority of the caffeine that was generated in the United States before to 1945. The
methylation agents that were utilized were dichloromethane and dimethyl sulfate (working,
2013).
Caffeine is a stimulant that acts on the central nervous system, and it is known to have a number

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of physiological effects on the body. It is a common ingredient in beverages such as coffee, tea,

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and soda, and it is also found in some medications and dietary supplements. It is generally

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considered safe for consumption in moderate amounts, but consuming large amounts of caffeine

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can lead to negative side effects (Pietrangelo, 2018).
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Caffeine is a purine alkaloid that belongs to the xanthine class of compounds. The structure of
caffeine consists of a nitrogen-containing ring, called a purine ring, which is fused to a xanthine
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ring. The purine ring contains two nitrogen atoms, and the xanthine ring contains one nitrogen
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atom and one oxygen atom. Caffeine also has four hydroxyl groups (-OH) attached to the
xanthine ring, and a methyl group (-CH3) attached to the nitrogen atom on the purine ring with a
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molecular weight of 194.19 g/mol (Xanthine, 2021).

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Figure 2: Caffeine Organic Structure

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How does the Separating Funnel Work?

To separate immiscible liquids, a separating funnel is used. Two
layers are visible when two immiscible liquids are put in a
separating funnel. Immiscible liquids are liquids that remain
homogeneous and don’t mix, in this experiment’s case, tea Tea Solution
mixture and dichloromethane are immiscible. The liquids are
poured into the funnel, and a stopcock or tap is used to control Dichloromethane
the flow of each liquid as it is drained out of the bottom of the
funnel. The purified liquids can be collected in separate
containers for further use. The lower layer will contain the
denser solvent (dichloromethane). The majority of halogenated
solvents are denser than water, but the majority of non-
halogenated solvents are less dense.

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Figure 3: Separating Funnel containing tea solution + dichloromethane

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2.1 Hypothesis

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Because I am familiar with all of the brands of tea I used in my experiment and because I tasted
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all four of them, I predict that the Tetley Black Tea will have the highest caffeine level compared
to the other varieties. This judgment is founded on the fact that I always have a heightened level
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of alertness and concentration after drinking this tea.

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3. Variables
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3.1 Independent Variable: Black Tea Brand (Tetley Black Tea, American Breakfast
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Black Tea, Filiz Black Tea, & Red Label Tea)

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The results of this experiment will simply depend on which brand of black tea includes the
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highest and lowest amounts of caffeine in order to establish which is the ideal beverage to
consume for each particular cause.
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3.2 Dependent Variable: Content/amount of caffeine extracted (in mg) per 5g of tea
Because the purpose of this experiment is to determine the quantity of caffeine that can be
extracted from 5 grams of each different brand of tea, this variable is entirely reliant on the
independent variable. Because of these results, the research question will be easily answered
using these results.

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3.3 Controlled Variables:

Controlled Variable How it’s Controlled Why it’s controlled

Measure exactly 200 ml of
water for every trial using a
Volume of water (ml) measuring cylinder.
Record each tea solution is at
35oC before adding
Temperature dichloromethane solvent in.
reacts with the tea and causes
Number of holes poked in the Poke exactly 32 holes around
For each 5g of tea leaves,
each will dissolve exactly in
200ml ensuring a fair
comparison between the
trials.
This is controlled because it
was discovered during the
experiment that the solvent
bubbles to form.
To guarantee that all

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aluminum foil covering the top of the foil covering evaporation ratio is the same
the petri dish. for all trials.

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Measure/weigh To ensure that a fair

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Amount of tea leaves from approximately 5g of tea experiment is conducted and

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each tea brand leaves.3@ caffeine content extracted is
fair.
Measure exactly 15ml of To ensure that most of the
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Amount of dichloromethane dichloromethane, adding the caffeine is extracted fully and

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added solvent and mixing it with the fairly from the tea mixture.
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tea 4 times.
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4. Apparatus/Equipment
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1. pipette
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2. 75mm funnel
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3. 250 ±5ml beaker

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4. 250 ±5ml conical flask

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5. 250 ± 0.1ml Graduated Cylinder

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6. 25 ±0.5ml measuring cylinder

7. 250ml separating funnel
8. Digital balance ±0.001g
9. Weighing boats
10. Petri dish
11. Sticker labels
12. Stirrer
13. Spatula spoon
14. 15ml dichloromethane
15. Ring stand
16. Thermometer
17. Aluminum foil
18. Filter Paper

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Separating Funnel
4.1 Photo of setup
Ring Clamp

Stand
Measuring Cylinder

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Funnel

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Conical Flask
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Beaker
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Dichloromethane
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4.2 Safety Precautions

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 Wear gloves while dealing with dichloromethane as it can cause several skin
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issues and irritations

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 Wear a mask for protection because dichloromethane causes several health issues
if inhaled such as headaches, light-headedness, agitation, and more
 Make sure to not pour dichloromethane when the tea solution is hot as it will form
a small reaction and produce a strong harmful smell

5. Procedure

1. Boil 200ml of distilled water

2. Weigh out approximately 5g of Tetley Black Tea leaves
3. Pour and stir the tea leaves in the beaker in the boiled water
4. Filter and separate the tea leaves from the tea solution into a 250ml conical flask using
filter paper and funnel

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5. Measure 15ml of dichloromethane into a 25ml measuring cylinder using a 1ml pipette
6. Prepare the separating funnel and place it on a ring stand and ensure the separating funnel
is closed
7. Wait until tea solution reaches 35oC and pour the solution into the 250ml separating
funnel
8. Pour 15ml dichloromethane into the tea solution in the separating funnel
9. Place a cork in the opening of the funnel and begin shaking and mixing the solution
together while releasing pressure every 10 seconds
10. Place a conical flask under the funnel and allow the dichloromethane mixed solution out
of the funnel slowly to avoid any tea entering and mixing with the solution
11. Repeat steps 8-10 3 more times
12. Label the petri dish with the brand of the tea and weigh it out
13. Pour the final dichloromethane solution into the labelled petri dish

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14. Cover the petri dish with aluminum foil and poke 32 holes for evaporation
15. Leave the solution to evaporate and caffeine crystalizes

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16. Weigh the petri dish and find the caffeine content by the difference in masses

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17. Repeat steps 1-16 with American Breakfast Black Tea, Filiz Black Tea, and Red Label

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Black Tea 3@
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6. Quantitative Data
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6.1 Raw Data

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5.000 (±0.001g) of Tea Leaves

Trial 1 Trial 2 Trial 3 Trial 4
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Tea Brand Mass of Mass of Mass of Mass of Mass of Mass of Mass of Mass of
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(Temperature 35 petri petri petri petri petri petri petri petri

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±0.5 oC) dish dish with dish dish with dish dish with dish dish with
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without caffeine without caffeine without caffeine without caffeine

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caffeine (±0.001g) caffeine (±0.001g) caffeine (±0.001g) caffeine (±0.001g)

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(±0.001g) (±0.001g) (±0.001g) (±0.001g)

Tetley Tea 37.121 37.148 37.121 37.149 37.121 37.146 37.121 37.147
American B. Tea 46.286 46.310 46.286 46.312 46.286 46.308 46.286 46.314
Filiz Tea 40.494 40.518 40.494 40.515 40.494 40.516 40.494 40.518
Red Label 34.451 34.475 34.451 34.474 34.451 34.475 34.451 34.476
Table 1: Raw Data and 4 trials of 4 different tea brands

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6.2 Processed Data

5.000 (±0.001g) of Tea Leaves
Tea Brand Trial 1 Trial 2 Trial 3 Trial 4 Mean of Standard Standard
(Temperature 35 (converted (converted (converted (converted each tea Deviation Error
±0.5 oC) from g to from g to from g to from g to across 4
mg) mg) mg) mg) trials
Tetley Tea 27 mg 28 mg 25 mg 26 mg 26.5 mg 1.12 0.56
American B. Tea 24 mg 26 mg 22 mg 28 mg 25 mg 2.24 1.12
Filiz Tea 24 mg 21 mg 22 mg 24 mg 22.75 mg 1.30 0.65
Red Label 24 mg 23 mg 24 mg 25 mg 24 mg 0.71 0.35
Table 2: Showing the mass of the caffeine content in each tea brand through trials 1, 2, 3, and 4, the Mean, Standard Deviation, and
Standard Error.

6.2.1 Sample Calculations:

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 Mean: Ex: Mean of Tetley Black Tea

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∑𝑿

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̅=
𝑿 ̅ = (𝟐𝟕+𝟐𝟖+𝟐𝟓+𝟐𝟔)

gm
𝑿
𝑵 3@ 𝟒

̅ = 𝟏𝟎𝟔 = 𝟐𝟔. 𝟓
𝑿
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𝟒
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x̄: Mean
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N: Total number of values

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Σx: Sum of terms

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 Standard Deviation
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Ex: Standard Deviation of Tetley

Black Tea

𝟏
𝝈 = √ ∑𝑵
𝒊=𝟏(𝒙𝒊 − 𝝁)
𝟐
𝑵

Where:
Xi: is an individual value
N: is the total number of values
μ: is the mean/expected value

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 Standard Error

𝜎
𝑆𝐸 = Ex: Standard Error of Tetley Black Tea
√𝑛
𝟏.𝟏𝟐
𝑺𝑬 = = 𝟎. 𝟓𝟔
√𝟒
Where:
SE: Standard Error Value
σ: sample standard deviation
√n: number of samples

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 Percentage Uncertainty

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𝑢𝑛𝑐𝑒𝑟𝑡𝑎𝑖𝑛𝑡𝑦
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𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑢𝑛𝑐𝑒𝑟𝑡𝑎𝑖𝑛𝑡𝑦 = × 100
𝑚𝑒𝑎𝑠𝑢𝑟𝑒𝑑 𝑣𝑎𝑙𝑢𝑒
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The formula used to determine the apparatus's % level of uncertainty is shown above. For
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instance, the mass of the petri dish with the caffeine solution for 5g of tea leaves in trial 1 taken
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from Tetley Black Tea has the following percentage uncertainty:

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0.001
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𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑢𝑛𝑐𝑒𝑟𝑡𝑎𝑖𝑛𝑡𝑦 = × 100 = 0.0027%

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37.148
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Tea Brand Uncertainty % Uncertainty of mass

(Digital Balance) obtained across 4 trials
Tetley Black Tea ±0.001 0.0027%
American Breakfast Black Tea ±0.001 0.0022%
Filiz Black Tea ±0.001 0.0025%
Red Label Black Tea ±0.001 0.0029%
Table 3: Percentage uncertainties obtained for the caffeine extracted from 5g of all types of Black Teas used during this experiment across 4
trials.

 Percentage Error
𝑚𝑒𝑎𝑠𝑢𝑟𝑒𝑑 𝑣𝑎𝑙𝑢𝑒 − 𝑡𝑟𝑢𝑒 𝑣𝑎𝑙𝑢𝑒
𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑒𝑟𝑟𝑜𝑟 = × 100
𝑡𝑟𝑢𝑒 𝑣𝑎𝑙𝑢𝑒

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The formula used to determine the % error is shown above using the measured value which was
obtained through the mean from the trials and the true value of the caffeine content equivalent to
5g of tea from the teas. Sample calculation of Tetley Black Tea is as follows:
26.5 − 27.8
𝑝𝑒𝑟𝑐𝑒𝑛𝑡𝑎𝑔𝑒 𝑒𝑟𝑟𝑜𝑟 = × 100 = −4.68%
27.8
Tea Brand % error obtained across 4
trials
Tetley Black Tea -4.68%
American Breakfast Black Tea -4.21%
Filiz Black Tea -5.02%
Red Label Black Tea -4.76%
Table 4: Percentage errors obtained for the caffeine content obtained from the experiment across 4 trials and the true values multiplied by 100.

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As the percentage error obtained for all teas is negative, we can say that the measured value is

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lower than the theoretical value. A negative percentage error means that there may have been

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some errors throughout the experimentation since the size of the error is such that the measured

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value is lower than the predicted value.
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 Bar Graph of the Tea Brand Mean

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Mean of each Tea Brand

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27 26.5
Mean of caffeine extracted (mg)

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25
25
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23 22.75
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21

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Tetley Tea American B. Tea Filiz Tea Red Label Tea
Tetley Tea Alokozay Tea Filiz Tea Red Label Tea

Brand of Black Tea

Mean

Graph 1: Graph showing an average mean of caffeine content extracted from 5g of each Tetley, American Breakfast, Filiz and Red Label Black
Teas.

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7. Qualitative Data
As the dichloromethane was being poured into the tea solution while it was at a higher
temperature, there was a brief period of bubbling and a strong smell from the
dichloromethane that could have been harmful. As a result, it was decided not to let the tea
solution sit out and cool down to a temperature lower than 40oC. The experiment had to be
redone a couple of times since some tea managed to escape the separating funnel, which
could have had an impact on how the caffeine crystallized and discovered its true mass.

8. Conclusion
As previously mentioned, the main aim of this experiment is to investigate the caffeine content
(mg) in different brands of black tea (Tetley Black Tea, American Breakfast Black Tea, Filiz

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Black Tea, & Red Label Tea) from dichloromethane using a separating funnel. According to the
findings of this investigation, Tetley Black Tea had the highest average caffeine content per 5g,

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at 26.5mg, while Filiz Black Tea had the lowest average caffeine content per 5g, at 22.75mg.

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The results show that some results were fairly comparable in terms of numbers and results,

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demonstrating the value of repeating the experiment and demonstrating how successful the trials
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were with few errors encountered. The dichloromethane bubbled and smelled dangerously as it
was poured into the tea solution at a higher temperature. Hence, the tea solution was not left to
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cool below 40oC. Some tea escaped the separating funnel, which could have affected caffeine
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crystallization and mass discovery. The experiment had to be repeated. In the end, the research
issue was well answered, and the mistakes that were made during the inquiry as well as the
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investigation itself provided valuable lessons. Since caffeine is a stimulant and can have a variety
of negative effects on the body, some individuals feel that it is crucial to keep track of how much
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of it they consume.
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9. Evaluation
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9.1 Strengths
1) To guarantee a fair test was conducted and to produce a credible test, this experiment was
repeated four times for each tea.
2) The same equipment and measurements were utilized throughout each and every test to
guarantee that the results were accurate and reliable.
3) During this experiment, a digital balance with an uncertainty of ±0.001g was employed.
This uncertainty is quite low and indicates that the number of errors was also very low.

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9.2 Weaknesses & Limitations

1) Because there was only one separating funnel capable of holding 250 milliliters that
worked, there was a shortage of lab equipment, which meant that the experiment could not be
completed and carried out in a more expedient manner.
2) It's possible that the amount of caffeine that was left in the petri dishes was altered by the
fact that some tea combination made its way into the sample of dichloromethane solvent
from the separating funnel.
3) Since each petri dish had a somewhat unique mass, the laboratory equipment was only
moderately dependable.
4) Percentage errors obtained were negative which concluded that human and experimental
errors occurred throughout.

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9.3 Improving the experiment

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Type of error Ways the errors could be Solutions
minimized or disregarded
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Misreading the measuring Ensure to read the

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cylinder when water is measuring cylinder at eye

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Random Errors poured where then the tea level so all solutions would
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solution would have have the same amount of

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different concentrations than water.

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the other tea solutions.

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There was only one 250ml Reduce the amount of tea

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separating funnel available solution used and

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for my experiment and the dichloromethane to match

tap was broken which
Systematic Errors allowed some of the tea
solution to enter the
dichloromethane where the
caffeine was supposed to be
extracted.
with the new lab separating
funnels available and ensure
to close the tap at the right
time to prevent the tea from
escaping into the petri dish.

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9.4 Extension
1) Increasing the scope of the inquiry and looking into the amount of caffeine found not only
in black tea but also in green, white, and oolong tea as well.
2) Find a new method to extract caffeine out of different tea brands such as using High
Performance Liquid Chromatography (HPLC).
3) Extracting caffeine from different solvents other than dichloromethane, that includes
solvents such as chloroform, ethanol, acetone and ethyl acetate with different methods such
as Soxhlet extraction, Ultrasonic extraction, and Heat Reflux extraction.

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10. References

Rich in antioxidants. (2020, August 24). The Times of India.

https://timesofindia.indiatimes.com/life-style/food-news/10-amazing-benefits-of-drinking-
black-tea/photostory/77671950.cms?picid=77671975
3D Model of caffeine. (2019, December 2). American Chemical Society.
https://www.acs.org/education/resources/undergraduate/chemistryincontext/interactives/brew
ing-and-chewing/3d-model-caffeine.html
Caffeine Structure, Formula & Composition | Caffeine Molecule Video. (2021). Caffeine

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Structure, Formula & Composition | Caffeine Molecule - Video & Lesson Transcript |

l.c
Study.com. Study.com. https://study.com/learn/lesson/caffeine-chemical-structure.html

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Calculator.net. (2019). Standard Deviation Calculator. Calculator.net.

gm
https://www.calculator.net/standard-deviation-calculator.html
3@
Dichloromethane - American Chemical Society. (2019, March 4). American Chemical
l1
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Society. https://www.acs.org/content/acs/en/molecule-of-the-
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week/archive/d/dichloromethane.html
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Mean. (n.d.). BIOLOGY for LIFE. https://www.biologyforlife.com/measures-of-central-

vs
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tendancy.html
dh

Pietrangelo, A. (2018, September 28). The Effects of Caffeine on Your Body. Healthline.
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https://www.healthline.com/health/caffeine-effects-on-body
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PubChem. (2019). Dichloromethane. Nih.gov; PubChem.

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https://pubchem.ncbi.nlm.nih.gov/compound/Dichloromethane
Standard Error in R (2 Example Codes) | User-Defined & std.error Function. (2019).
Statistics Globe. https://statisticsglobe.com/standard-error-in-r-example
Working, I. (2013). Caffeine. Nih.gov; International Agency for Research on Cancer.
https://www.ncbi.nlm.nih.gov/books/NBK507027/#:~:text=Synthetic%20production%20of%
20caffeine%20involves
Xanthine. (n.d.). Go.drugbank.com. Retrieved December 31, 2022, from
https://go.drugbank.com/drugs/DB02134

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