Determination of caffeine content in tea

SUBMITTED BY
NAME: HARI HARA SUDHAN ,
ADAM SAM RAJ,
MAHAREESH BALA
CLASS AND SECTION: XII-A

1
 INDEX

S.NO TOPIC PG.NO

1. Introduction 3

2. Purpose 6

3. Aim 6

4. Materials required 6

5. Procedure 7

6. Observations 8

7. Inference 9

8. Reference 9

2
Introduction:
Tea is one of the most widely consumed beverages globally, valued
for its unique flavors, calming properties, and potential health
benefits. It is derived from the leaves of the Camellia sinensis plant,
which undergo various processing methods to produce different
types of tea, such as black tea, green tea, oolong tea, and white tea.
Alongside the numerous bioactive compounds present in tea,
caffeine is one of the prominent alkaloids that contributes to its
stimulating effects.

Caffeine, a natural compound belonging to the class of alkaloids, is

widely known for its ability to stimulate the central nervous system.
It is a psychoactive substance that provides temporary alertness and
improved cognitive performance. Caffeine is present in various plant
sources, including coffee beans, cocoa beans, and tea leaves. Among
these, tea is an important dietary source of caffeine, and
understanding its caffeine content is of interest to both researchers
and consumers.

The determination of caffeine content in tea holds significant

relevance for several reasons. Firstly, it provides valuable
information for individuals who are conscious of their caffeine
intake. Many people monitor their caffeine consumption due to
personal preferences, health concerns, or the need to limit their
intake. By knowing the caffeine content in tea, individuals can make
informed decisions about their beverage choices and adjust their
consumption accordingly.

Furthermore, variations in caffeine levels among different tea types

and brands can have implications for consumers. The effects of
caffeine on the body can vary depending on its concentration and
consumption patterns. Some individuals may prefer teas with lower
caffeine content to avoid potential side effects, while others may
seek teas with higher caffeine levels for a more stimulating effect.
Additionally, regulatory bodies may set limits on caffeine content in

3
certain products, and determining the caffeine levels in tea can
ensure compliance with these regulations.

The objective of this project is to quantitatively determine the

caffeine content in different tea samples using a chemical extraction
and spectrophotometric method. By analyzing and comparing the
caffeine levels in various teas, we aim to gain insights into the
variations in caffeine content among different tea types and their
potential implications.

The caffeine content in tea can vary due to several factors. Firstly,
the type of tea plays a significant role in determining caffeine levels.
For example, black tea generally contains higher levels of caffeine
compared to green tea due to differences in processing methods.
The oxidation process involved in producing black tea allows for the
retention of more caffeine compared to the less oxidized green tea.

Other factors that influence caffeine content in tea include the

geographical origin of the tea leaves and the specific tea cultivar.
Different regions and cultivars may exhibit variations in caffeine
levels due to environmental factors and genetic differences.
Additionally, brewing parameters, such as water temperature and
steeping time, can affect the caffeine extraction efficiency during tea
preparation.

To analyze the caffeine content in tea, a reliable and accurate

method is required. Various techniques have been employed for
caffeine extraction and quantification, including chromatographic
methods such as high-performance liquid chromatography (HPLC)
and gas chromatography (GC), as well as spectrophotometric
methods. In this project, we will utilize a spectrophotometric method
based on the measurement of absorbance using a UV-Vis
spectrophotometer.

The spectrophotometric method for caffeine determination relies on

the principle that caffeine exhibits maximum absorbance at a specific

4
wavelength in the ultraviolet-visible (UV-Vis) range. By measuring the
absorbance of the caffeine solution at this wavelength, we can
correlate it with the caffeine concentration using a calibration curve.
The calibration curve is prepared by analyzing known concentrations
of caffeine standards.

In conclusion, the determination of caffeine content in tea is of great

importance for understanding its variations among different tea
types and brands. This project aims to provide quantitative data on
the caffeine levels in tea samples using a chemical extraction and
spectrophotometric method. By investigating the caffeine content in
various teas, we can gain valuable insights into the caffeine
variability and help individuals make informed choices about their
tea consumption based on their desired caffeine levels or personal
preferences.

Purpose:

5
The purpose of this project is to quantitatively determine the
caffeine content in various tea samples using an extraction and
spectrophotometric method. By analyzing and comparing the
caffeine levels in different teas, we aim to gain a better
understanding of the variations in caffeine content and their
potential implications.

Aim:
The aim of this project is to investigate and compare the caffeine
content in different tea samples, including black, green, oolong, and
white tea. By conducting this analysis, we seek to identify any
significant differences in caffeine concentrations among these tea
types and draw conclusions based on the obtained data.

Materials Required:

 Tea samples
 Analytical balance
 Mortar and pestle
 Solvent (e.g., methylene chloride or ethyl acetate)
 Separatory funnel
 Distillation apparatus
 Rotary evaporator
 Analytical balance
 UV-Vis spectrophotometer
 Caffeine standard solution
 Glassware (beakers, flasks, pipettes, etc.)
 Safety goggles, gloves, and lab coat

Procedure:

6
1. Sample Collection and Preparation: In this project, several
samples of different tea types will be collected. These samples
can be obtained from local tea stores or online retailers. Once
the samples are collected, they need to be prepared for
analysis. Approximately 5 grams of each tea sample will be
weighed using an analytical balance. The tea leaves will then be
ground into a fine powder using a mortar and pestle to ensure
uniformity.
2. Caffeine Extraction: To extract caffeine from the tea samples, a
suitable solvent such as methylene chloride or ethyl acetate
will be used. The powdered tea sample will be transferred to a
beaker, and the solvent will be added to immerse the tea
completely. The mixture will be stirred gently for about 15
minutes to facilitate the extraction process. Subsequently, the
liquid extract will be separated from the solid residue by
filtration using filter paper.
3. Isolation of Caffeine: The liquid extract obtained from the
previous step will be transferred to a separatory funnel. An
equal volume of water will be added to the funnel, and the
mixture will be gently shaken. This step aims to separate the
organic layer (containing caffeine) from the aqueous layer. The
lower organic layer will be drained into a separate flask while
discarding the aqueous layer. This process will be repeated with
the remaining tea samples.
4. Purification: The extracted caffeine solution will still contain
some solvent. To obtain pure caffeine, the solvent needs to be
removed. This will be achieved by setting up a distillation
apparatus. The extracted solution will be heated at a controlled
temperature, allowing the solvent to evaporate. The solvent
vapors will condense and be collected separately, leaving
behind purified caffeine.

5. Caffeine Quantification: A caffeine standard solution will be

prepared by dissolving a known amount of caffeine in a suitable
solvent, such as methylene chloride. This standard solution will

7
 serve as a reference to determine the caffeine content in the
tea samples. The absorbance of the standard solution will be
measured using a UV-Vis spectrophotometer at a specific
wavelength, such as 272 nm. This will create a calibration curve
relating the absorbance to the concentration of caffeine.
Next, the absorbance of the extracted caffeine solutions
obtained from the different tea samples will be measured using
the same wavelength. By comparing the absorbance values of
the tea samples with the calibration curve, the concentration of
caffeine in each sample can be determined.

Observation:

Weight of Tea Volume of Absorbance at Caffeine Concentration

Sample Leaves (g) Solvent (mL) λmax (mg/mL)

Tea A

Tea B

Tea C

Tea D

Inference:

8
 Sample Caffeine Content (mg/mL) Inference

Tea A Tea A has a ___________________caffeine content.

Tea B Tea B has a _____________________ caffeine content.

Tea C Tea C has a _____________________ caffeine content.

Tea D Tea D has a _____________________caffeine content.

Reference:

https://www.agriculturejournals.cz

https://www.teatulia.com

https://www.healthline.com

https://www.betterhealth.vic.gov.au