CHM 580 Spectrochemical Methods of Analysis Laboratory Report Experiment 2: Ultraviolet-Visible Spectros
CHM 580 Spectrochemical Methods of Analysis Laboratory Report Experiment 2: Ultraviolet-Visible Spectros
CHM 580 Spectrochemical Methods of Analysis Laboratory Report Experiment 2: Ultraviolet-Visible Spectros
2. ABSTRACT
The sample used in this experiment is a bag of tea where the caffeine was
extracted from it. The sample was determined by using the Ultraviolet-visible
spectrosocopy to know the concentration and the amount of caffeine in the sample. The
data was analysed by using background-correction method and second derivative. The
sample has been triplicate to ensure the data more accurate and precise. The samples
were diluted with dilution factor of 10 to ensure the amount of caffeine in samples is in
the standard range. The standard solutions of 10ppm, 20ppm, 30ppm, 40ppm and 50ppm
was prepared from 100ppm using dilution technique. The concentration of the three
samples were 10.80 ppm, 10.0 ppm and 10.85 ppm with the relative standard deviation of
2.2361. The amount of the caffeine in the sample 1, sample 2 and sample 3 was
0.027000g , 0.027125g and 0.0271125 respectively with the confidence limit ± 0.9232 g.
The weight percent was 1.09% for sample 1 and 3 while 1.10% for sample 2.
3. INTRODUCTION
Caffeine can have both positive and negative health effects. It can treat and
prevent the premature infant breathing disorders. Some people experience sleep
disruption or anxiety if consume caffeine. affeine can produce a mild form of drug
dependence which associated with withdrawal symptoms such as sleepiness, headache,
and irritability.
Sodim hydroxide solution was prepared by adding 0.4g of solid hydroxide into a
10ml of volumetric flask and diluted with distilled water up to the calibration mark.
For the analysis, eight centrifudges were used. The tubes consists five standards
solution and three sample solution. Each tube was filled with 1ml of sodium
hydroxide, 4 ml of copper(II) acetate solution and 5 ml of sample tea solution. The
solutions were analysed by UV Spectrophotometry Lambda 35.
6. RESULTS
Table 1 : The amount of caffeine in teabag using second derivatives ultraviolet spectroscopy
The absorption spectra of caffeine in tea bag was measured second derivative
using UV-visible spectrometer. The second derivative of caffeine was measured at
298.6nm. There are three preparations required for this experiment which is sample
solution preparations, tea sample preparations and copper (II) acetate solutions. The
concentration of caffein in the test samples was determined by the calibration curve and
comparison with the standard solution of caffeine. Calibration graph was obtained
between the absorbance intensity on y–axis and concentration of caffeine (mg/L) on x–
axis followed Beer-Lambert’s law over the concentration range of standard solution
measured 0.0-50.0 ppm.
From the graph of second derivative of absorbance versus the wavelength, the
result for standard 1 until standard 5 and sample 1, sample and sample 3 was obtained.
The method of linear regression was used for data evaluation and to determine the
concentration of sample 1, sample 2 and sample 3. The graph representing the second
derivative of absorbance versus the concentration of standard was plotted. The
concentration of sample 1, sample 2 and sample 3 are 10.80ppm, 10.90ppm and
10.85ppm respectively. Hence, by using the suitable formula, the original amount of
caffeine in tea bag in 250ml was obtained. The amount of caffeine for sample 1, sample
2 and sample 3 are 27.00 mg, 27.25mg and 27.125mg respectively. The reading was far
away from expected. The weight percent (w/w%) of were calculated. From calculation,
weight percent of sample 1 and sample 3 was 1.09% while sample 2 was 1.10%. The
value of weight percent is lower because of the amount of caffeine in the samples was out
of the range from the original amount of caffeine.
Precision of the method was determined by three replications of each sample. The
precision (%RSD) of the replications should less than two which is indicative of a precise
method. The smaller the value of relative standard deviation to xero indicates the more
accurate sample preparation. But, the relative standard deviation of this experiment is
higher than two which is 2.2361. Therefore, there is errors when doing this experiment or
the preparations of sample.
The concentaration of the sample 1, sample 2 and sample 3 were 10.80 ppm, 10.0 ppm
and 10.85 ppm respectively with the relative standard deviation of 2.2361. The amount of
the caffeine in the sample 1, sample 2 and sample 3 was 0.027000g , 0.027125g and
0.0271125 respectively with the confidence limit ± 0.9232 g. The weight percent was
1.09% for sample 1 and 3 while 1.10% for sample 2.
9. REFERENCES
Goodwin, L. (2019, March 26). How Much Caffeine Is in Your Drink of Choice?
Retrieved April 7, 2019, from https://www.thespruceeats.com/caffeine-in-coffee-
tea-cola-765276
Marina, S. (n.d.). UV-VIS SPECTROPHOTOMETRIC TRACE LEVEL
DETERMINATION OF CAFFEINE IN 'RED BULL'. Retrieved April 7, 2019,
from https://www.academia.edu/9264278/UV-
VIS_SPECTROPHOTOMETRIC_TRACE_LEVEL_DETERMINATION_OF_C
AFFEINE_IN_RED_BULL?auto=download
Caffeine. (2019, April 03). Retrieved April 7, 2019, from
https://en.wikipedia.org/wiki/Caffeine
A., Azhar. (n.d.). UV-Visible Spectroscopy. Retrieved April 7, 2019, from
https://www.scribd.com/document/267919454/UV-Visible-Spectroscopy
10. QUESTION
1. Write the wavelength ranges for the ultraviolet and visible region of the
spectrum?
Ultraviolet : 180nm-380nm
Visible : 380nm-780nm
2. List the molecular or structural features that give rise to absorption of the
ultraviolet/visible (UV/VIS) radiation in organic species. Give specific sample
example of an organic compound that would not absorb UV/VIS radiation.
Double and triple bonds and those bond involve the pi bond was absorb
UV/VIS radiation. For example, alkene, alkyne and aromatic compound.
Single bond such as carbon and hydrogen in alkanes and those are sigma
bonds does not absorb UV/VIS radiation. For example is hexane C6H6
3. Why is it important to determine caffeine in tea using background correction
and second derivative ultraviolet-visible spectrophotometry?
To remove the interference in the absorption wavelength that may cause by other
substances such as tannis that maybe found in tea leaves. Therefore, the second
derivative is required in this experiment. To minimize the tannis the sodium
hydroxide is used.