TITLE

Determination of Caffeine in Tea by Using Second Derivative Ultraviolet-Visible Spectroscopy

ABSTRACT
The main purpose of this experiment is to determine the amount of caffeine in percent
weight in tea samples using background-correction method and second derivative Ultravioletvisible spectrophotometry. Background-correction methods were used to remove the
interferences on the reading and the second derivative for caffeine were measured at 298.6nm. 2g
tea samples were boiled to extract the compounds in the tea sample and the standard solutions of
10 ppm, 20 ppm, 30 ppm, 40 ppm and 50 ppm were prepared from 100ppm by using dilution
technique. 3 samples of caffeine were prepared from the boiled sample. Then, the samples and
standard solutions were analysed with a blank sample using second derivatives of UV
spectrophotometry. Copper acetate and sodium hydroxide were added into the standard and
sample solutions. The instrument used is Ultraviolet-visible spectrometer Perkin Elmer Lambda
35.

INTRODUCTION
Caffeine is the common name for 1,3,7-trimethylxanthine. When purified, caffeine
produces an intensely bitter white powder that provides a distinctive taste in soft drinks. The
word "caffeine" came from the German word kaffee and the French word caf, each meaning
coffee. Caffeine is a drug that is naturally produced in the leaves and seeds of many plants. It's
also produced artificially and added to certain foods. Caffeine is defined as a drug because it
stimulates the central nervous system, causing increased alertness. In fact, caffeine itself is a mild
painkiller and increases the effectiveness of other pain relievers. Caffeine gives most people a
temporary energy boost and elevates mood. Caffeine is in tea, coffee, chocolate, many soft
drinks, and pain relievers and other over-the-counter medications. In its natural form, caffeine
tastes very bitter. But most caffeinated drinks have gone through enough processing to
camouflage the bitter taste. Caffeine content can range from as much as 160 milligrams in some
energy drinks to as little as 4 milligrams in a 1-ounce serving of chocolate-flavored syrup. Even
decaffeinated coffee isn't completely free of caffeine. These products can contain as little as 16
milligrams or as much as 200 milligrams of caffeine. According to the U.S. Food and Drug
Administration (FDA), about 90 percent of the worlds population ingests some form of caffeine.
In the United States, about 80 percent of us take in some caffeine every day. According to
the Mayo Clinic, its safe for most healthy adults to consume up to 400 milligrams of caffeine
per day. How much caffeine each beverage contains varies a lot, so reading labels is important.
Theres also a great variation in the amount of caffeine an individual can tolerate without
unpleasant side effects. If you consume roughly the same amount of caffeine every day, you can
develop a tolerance to it. Your tolerance level is affected by your age, body mass, and any health
conditions you may have. The hypothesis for this experiment is

METHODOLOGY
A.Preparation of samples solutions
2g of tea sample weighed was boiled with 200ml of distilled water on hot plate for 15
minute. Then cooled the sample solution at room temperature filtered using Buchner funnel and
diluted it with distilled water in 250ml volumetric flask until calibration mark. Finally, the
sample solution is diluted again by adding 5 mL of tea sample prepared into a 50 mL volumetric
flask and then diluted up to the calibrated mark.
B.Preparation of standard solution
From 100ppm caffeine stock solution, standard solutions of 10ppm, 20ppm, 30ppm,
40ppm, and 50ppm were prepared. Then each of it was diluted with distilled water until the
calibration mark in a 50mL volumetric flask. The formula of C1V1 = C2V2 was applied for the
dilution.
C.Preparation of copper (II) acetate
After that, 0.025 M of copper (II) acetate solution is prepared by adding 0.25g of copper
(II) acetate anhydrous into a 50 mL volumetric flask and diluted with distilled water up to the
calibration mark. And 1.0 M of sodium hydroxide solution is prepared by adding 2 g of solid
hydroxide into a 50 mL volumetric flask and diluted with distilled water up to the calibration
mark.

For this analysis, nine centrifuges tubes were used. The tubes consist of 6 standard
solutions and 3 sample solutions. Each tube was filled with 4.0ml of 0.025M copper (II) acetate
solution and 1mL of 1.0M sodium hydroxide. The tubes were shaken in centrifuge for 10
minutes. Finally, the solutions are analysed by UV spectrophotometry Lambda 35.

RESULT
Sample

Concentration of
Second
Weight (g)
% weight
sample ( mg/L)
deravative
1
31.5
0.1872
0.1575
6.66
2
29.5
0.16902
0.1475
6.24
3
31.0
0.1875001
0.1650
6.98
Table 1: The amount of caffeine in teabag using second derivatives ultraviolet spectroscopy

Upper
Limit
7.54945

DISCUSSION

Lower
Standard deviation,
Relative standard
Limit
sd
deviation
5.71055
0.3701
5.58%
Table 2: Statistical value of caffeine sample

The aim of this experiment is to determine the amount of caffeine in percent weight in tea
samples using background-correction method and second derivative Ultraviolet-visible
spectrophotometry. The caffeine was measured at 298.60nm. Second derivative is used in this
experiment is to remove the interference in the absorption wavelength. That may cause by other
substance such as tannins that maybe found in tea leaves. the second derivative is required in this
experiment. There are three preparations required for this experiment which is sample solutions
preparations, tea samples preparations and copper (II) acetate preparations. Five concentrations
of 10ppm, 20ppm, 30ppm, 40ppm, and 50ppm of stock solution were prepared from 100ppm
caffeine. Then, copper(II) acetate and sodium hydroxide were prepared. Adding 0.1 M NaOH
and 0.025 M copper (II) acetate into the samples will help in suppressing the other substances
(tannins) in the solutions.

All solutions were diluted up to 50ml calibration mark using

C1V1=C2V2 formula. Nine solutions consist of standard solutions and sample solutions were
centrifuged. From the result obtained, the graph of second derivative versus caffeine
concentration was plotted . The second derivative of standard 1, standard 2, standard 3, standard
4 and standard 5 are 0.0598, 0.1171, 0.1758, 0.2296 and 0.2854 respectively. Hence, using the
appropriate formula, the actual weight of sample and weight percent (%w/w) were calculated.
From calculation, the actual amount of caffeine in tea sample are 0.1575g, 0.1475g, and 0.1650g.
for sample 1, sample 2 and sample 3 respectively. These samples have concentration of 31.5ppm,
29.5ppm and 33.0ppm. In this experiment we need to take some precaution step during carry out
the experiment. Firstly, use the pipette in the correct way because, it will affect the volume of
sample that we take that lead to different concentration.. Beside, make sure the time take to
centrifuge the sample are enough to make sure all the solid was precipitate. The lipton tea found
safe to be consumed by people

CONCLUSION

In conclusion, the tea sample have concentration of 31.5ppm, 29.5ppm and 33ppm. The
average amount of tea sample in lipton tea is 0.1570g and habe weight percent of 6.63%.

REFERENCES
Seedher, N, and Sidhu, K., (2007). Studies on the use of tea leaves as pharmaceutical adsorbent.
International journal of biological chemistry 1 (3): 162-167. Retrieve on 2015, May 22 nd from
http://scialert.net/qredirect.php?doi=ijbc.2007.162.167&linkid=pdf.
Alipoor, B., and Rad A.H. (2012). A Review on the Therapeutical Effects of Tea. Asian Journal
of Clinical Nutrition 4 (1): 1-15. Retrieve on 2015, June 15 from http://scialert.net/qredirect.php?
doi=ajcn.2012.1.15&linkid=pdf.
Dash, S., S., and Gummadi S., N., (2008). Inhibitory Effect of Caffeine on Growth of Various
Bacterial Strains. Rearch Journal of Microbiology 3 (6): 457 465. Retrieve on 2015, May 22 nd
from http://scialert.net/qredirect.php?doi=jm.2008.457.465&linkid=pdf.

CHM 580
SPECTROCHEMICAL METHODS
TITLE: DETERMINATION OF CAFFEINE IN
TEA SAMPLES (LIPTON TEA)
USING SECOND-DERIVATIVE UV
SPECTROSCOPY
NAME
ID NO.
CLASS
PARTNERS

: AIDA NURUL FATIHAH BINTI MAJURI

: 2014824842
: AS245 4D1
: NUR KHAIRUNNISA BINTI ROSNAN 2014824842
NUR LIYANA BINTI MISRAN
2014632744

DATE OF
SUBMISSION : 21/12/2015
LECTURER : MADAM ZARILA BT MOHD SHARIFF