DETERMINATION OF CAFFEINE IN A TEA SAMPLE
An Investigatory Project (Class XII - CBSE Format)
Submitted to: ____________________________
Name: _______________________
Class: 12 Roll No.: _____ School: ___________________
Guide: _______________________
Date: _______________________
Class XII - CBSE Investigatory Project
AIM & OBJECTIVES
AIM
To determine the percentage of caffeine present in a given sample of tea using UV-
Visible spectrophotometry and a calibration curve.
OBJECTIVES
1. To prepare standard solutions of caffeine and construct a calibration curve. 2. To
measure the absorbance of the tea extract and determine its caffeine concentration
using the calibration graph. 3. To calculate the percentage of caffeine in the tea
sample and discuss sources of error.
INTRODUCTION
INTRODUCTION
Caffeine (1,3,7-trimethylxanthine) is a natural stimulant found in tea leaves.
Quantitative determination of caffeine is commonly performed using UV-Visible
spectrophotometry after appropriate extraction and dilution. The method involves
preparing standard solutions of known caffeine concentration, measuring their
absorbances, constructing a calibration graph (Absorbance vs Concentration), and
using the line of best fit to determine the unknown concentration in the tea sample.
APPARATUS & REAGENTS
APPARATUS AND INSTRUMENTS
1. UV-Visible spectrophotometer (or colorimeter) with 1 cm quartz cuvette. 2.
Analytical balance (sensitivity 0.001 g). 3. Volumetric flasks (10 mL, 25 mL, 50 mL,
100 mL). 4. Pipettes and micropipettes. 5. Beakers, conical flasks, filter paper,
separating funnel (if liquid-liquid extraction used).
REAGENTS
1. Pure caffeine standard. 2. Distilled water. 3. Solvent for extraction (e.g.,
dichloromethane or ethyl acetate) if extraction method used. 4. Standard laboratory
reagents as required for any specific extraction or cleanup steps.
PROCEDURE
PROCEDURE (Summary)
1. Prepare a stock solution of caffeine (1000 µg/mL) by dissolving accurately weighed
caffeine in distilled water or suitable solvent and dilute to volume in a volumetric
flask.
2. Prepare standard solutions of concentrations 0, 20, 40, 60, 80, and 100 µg/mL by
serial dilution of the stock solution.
3. Measure the absorbance of each standard at the wavelength of maximum absorption
for caffeine (typically around 273 nm) using a blank to zero the spectrophotometer.
4. Plot a calibration curve of absorbance (y-axis) vs concentration (x-axis) and
determine the equation of the best-fit line.
5. Prepare the tea sample: weigh 1.000 g of finely powdered tea, extract caffeine
with suitable solvent, filter, and dilute the final extract to 100 mL.
6. Measure the absorbance of the prepared tea extract under the same instrument
settings and use the calibration equation to find the concentration of caffeine in
the extract.
7. Calculate mass of caffeine and percentage (%) caffeine in the tea sample.
OBSERVATION TABLE (Calibration Standards)
Standard No. Concentration (µg/mL) Absorbance (A)
1 0.0 0.000
2 20.0 0.112
3 40.0 0.225
4 60.0 0.341
5 80.0 0.455
6 100.0 0.569
Calibration Curve: Caffeine (Absorbance vs Concentration)
A = 0.005700 C + -0.001333
0.6
0.5
0.4
Absorbance (A)
Sample
(52.5 µg/mL, 0.298)
0.3
0.2
0.1
0.0
0 20 40 60 80 100
Concentration (µg/mL)
CALCULATIONS & RESULTS
CALCULATIONS & RESULTS
Linear regression (best-fit line) from calibration data: A = m*C + b
Calculated slope (m) = 0.005700 absorbance units per µg/mL
Calculated intercept (b) = -0.001333 absorbance units
Measured absorbance of tea sample: A_sample = 0.298
Concentration of caffeine in final solution: C_sample = (A_sample -
b)/m = (0.298 - -0.001333)/0.005700 = 52.515 µg/mL
Total mass of caffeine in 100 mL solution = 52.515 µg/mL × 100 mL =
5251.462 µg = 5.251 mg
Percentage of caffeine in 1.000 g tea sample = 0.5251 % w/w
PLACEHOLDER DIAGRAMS
Spectrophotometer
Beaker
(sample)
Hand-drawn style placeholder for extraction setup and instrument connection
CONCLUSION
CONCLUSION
The percentage of caffeine in the tested tea sample was found to be 0.5251 % (w/w)
based on the example data and assumptions provided. Results will vary based on
extraction efficiency and instrument precision.
PRECAUTIONS
PRECAUTIONS
1. Use calibrated glassware and pipettes for accurate volume measurements.
2. Ensure the spectrophotometer is properly zeroed using the blank at the chosen
wavelength.
3. Filter extracts to remove particulates that may scatter light.
4. Work quickly and avoid exposure of samples to sunlight if using light-sensitive
reagents.
5. Repeat measurements (triplicate) for better precision.
BIBLIOGRAPHY
BIBLIOGRAPHY / REFERENCES
1. Skoog, D.A., Holler, F.J., & Crouch, S.R. (2014). Principles of Instrumental
Analysis.
2. AOAC Official Methods of Analysis methods on caffeine determination.
3. Standard laboratory manuals and supplier instructions for spectrophotometer
operation.