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INTRODUCTION:
Even among adults, heavy caffeine use can cause unpleasant side effects. Caffeine are not
good for those who take certain medications. Heavy daily caffeine use more than 500 to
600mg a day may cause side effects such as insomnia, nervousness, restlessness, irritability,
stomach upset, fast heartbeat and muscle tremors. Some people may be more sensitive to
caffeine than others. If you are susceptible to the side effects of caffeine, just small amounts
of it may prompt unwanted effects, for example sleep problems. How you react with caffeine
may be determined in part of how much caffeine you consumed. People who did not take
caffeine regularly tend to be more sensitive to its negative effects. Others factors include
body mass, age, medication use and health conditions such as anxiety disorder. Research also
suggest that men may be more susceptible to the effects of caffeine than women.
Caffeine works by stimulating the central nervous system (CNS), heart, muscles and the
centre that control blood pressure. Caffeine can also rise the blood pressure, but might not
have this effect in people who are used to it. Caffeine can also act like a water pill thats
increase the urine flow. But again, it may not have this effect in people who consume caffeine
regularly. Also, drinking coffee during moderate exercise is not likely to cause dehydration.
UV-VIS Molecular Absorption Spectrometry is the most common analytical technique in
the analytical laboratory in which involves the absorption of ultraviolet and visible radiation
for quantitative purposes. Absorptions commonly occurs with many organic molecules,
metals and metal-organics complexes. In UV-VIS, absorption involves bonding (outer
valence) electrons. UV-VIS spectrophotometer are made up of stable source of radiant
energy, a transparent sample container, a device for isolating specific wavelength, a radiation
detector which converts transmitted radiation into a useable signal and a signal processor or
readout. UV-VIS spectra can be used to detect the presence of absorbing functional groups or
chromophores.
UV-VIS spectroscopy is valid, simple and cost effective method in determining the
concentrations of absorbing species if applied to pure compounds and used with the
appropriate standard curve. A standard curve relating absorbance to concentration can be
developed for any compounds and used to determine the concentration of sample containing
the same compound. The analysis should be done at a wavelength of maximum absorbance
and located in relatively flat region of the spectra. This is to ensure that the absorbance will
be high and constant in narrow range around the chosen wavelength. The optimal wavelength
should be selected on the good absorbance of the analyte and low absorbance by other
species presence in the solution. This wavelength will allow valid absorption measurements
to be made on analyte samples that contain mixture of materials.
The sample container, that we called cuvette or cell, must be constructed of a material that
is transparent to radiation in the wavelength range of interest. Containers of quartz or fused
silica are necessary in the UV range and can be used in the 700 -3000nm infrared region.
Glass and plastic can be used in the visible region as well. The wavelength selector are filters
and monochromators. The detector are phototube, photomultiplier tube (PMT), photodiode,
photodiode array.
RESULT AND CALCULATIONS:
V1 = 0.25 ml
V1 = 0.50 ml
V1 = 0.75 ml
V1 = 1.00 ml
V1 = 1.25 ml
0.8
Absorbance 0.6
0.4
0.2
0
0 20 40 60 80 100 120 140 160
Concentration
y= mx + c where;
Tea sample 1:
Y = mx + c
X = 48.630 ppm
Tea sample 2:
Y = mx + c
X = 20.851 ppm
Tea sample 3:
Y = mx + c
X = 39.370 ppm
Calculation for the tea sample molar absorptivity using the equation:
A = bc where;
A = absorbance, = molar absorptivity, b = length of cuvette, c = concentration of analyte
Tea sample 1:
0.130 = (1.00) (48.630)
= 2.67 x 10-3
Tea sample 2:
0.055 = (1.00) (20.851)
= 2.64 x 10-3
Tea sample 3:
0.105 = (1.00) (39.370)
= 2.66 x 10-3
DISCUSSION:
In this experiment, the spectrum for standard caffeine and caffeine from the tea sample were
obtained. The maximum peak of absorption was measured at peak 285nm, this is because the max
is the wavelength that gives the maximum value applied in the experiment. The peak
maximum correspond to the maximum value of molar absorptivity and hence the sensitivity
of the analysis.
Beers Law typically applied in this experiment. Beers law is strictly valid only where the
molar absorptivity changes little wavelength. The linearity of the Beers Law is limited to
chemical and instrumental factors. Beers law describe absorption behaviour of relatively low
analyte concentration in a solution as a limiting law. At high concentration will involve
solute-solvent interaction, solute-solute interaction and hydrogen bonding which will affect
the analyte concentration and its absorptivity. Beers law applies monochromatic radiation
with use of continuous source and a monochromator which will provide a very narrow band
with an effective bandwidth. Therefore, the selection of the wavelength should be made at the
max.
In this experiment, we can observed that there are few chromophores presence in the
caffeine molecule by the presence of carbonyl group (C=O) in it. A chromophore is the
functional group of an organic molecule that responsible for the molecule to be coloured.
This particular chromophore will undergo an n * transition around 280-290 nm.
Therefore, by this fact it is clearly stated why the peak of 285nm wavelength was chosen as
the taken into account. Secondly, there were possibilities that the structure undergo - *
transition. This energy transition was due to the presence of double bond and will occur in the
near UV region. According to Pavia, Lampman, Kriz and Vyvyan (2015), the energy
happened in this type of transition is rather a high energy level and their positions are
sensitive to the presence of substitution.
The absorbance values that we obtained from the spectrum of standard caffeine was 0.019,
0.055, 0.085, 0.105 and 0.130 according to their respective concentration. Meanwhile, for the
sample we analysed, the absorbance value that take was taken are 0.130, 0.055 and 0.105
respectively. After calculations had been made, we obtained the percentage weight (w/w)
with the value of 9.07%. This came from the result in plotting the Beers law graph to figure
out the quantitative value of the caffeine from the tea samples we had prepared.
The experiment been made, but we cannot said it is totally succeed even though from the
graph we obtain, the R2 = 0.9926 is nearly to 1. This is because, the sample container or the
cuvette is not matched and it is not clear and transparent to let the light to penetrate through it
completely. Other than that, to get a good result of the Beers law plot, the concentration of
the sample analysed must be diluted to become a sample solution that is very low
concentration. If the solution contains many solute, it could lead to a greater scatter effect of
the source of radiation. In liquids, the extinction coefficient will usually changes slowly with
the wavelength. The rate of change in absorbance with wavelength is the smallest at the
maximum peak of absorbance curve. Therefore, the measurement will be made at this
particular peak to minimize the errors produced by errors in wavelength in the instruments
that is errors due to having a different extinction coefficient than assumed.
CONCLUSION:
Caffeine in the tea sample was successful analyse by using UV-VIS
spectrophotometer. Next, the concentration of caffeine in the tea sample was
successful determined by graph. The average concentration of caffeine in
triplicate sample is 362.84ppm and the amount of caffeine presence in it is
181.42mg. The Beers law equation for this analysis is y = 0.0027x + 0.0013.
REFERENCES:
1. Skoog, D.A., Holler, F.J. & Crouch, S.R. (2007). Principle of Instrumental
Analysis
(6th Edition). Belmont, CA: Brooks/Cole, Cengage learning.
4. Bhawani, S.A., Fong, S.S., & Mohamad Ibrahim, M.N. (2015). Spectrometric
Analysis of Caffeine. International Journal of Analytical Chemistry.
Retrived from http://dx.doi.org/10.1155/2015/170239.
5. Pavia, D.L., Lampman, G.M., Kriz, G.S., & Vyvyan, J.R. (2015). Introduction to
Spectroscopy. (5th Edition). Singapore: Cengage learning Asia Pte Ltd.
QUESTIONS:
Leaving the tea bag longer will make a stronger cup of tea. The concentration of
caffeine (along with the flavour molecules and everything else) will slowly trend
towards an equal concentration in the leaf and in the water. The longer the tea bag in
hot water, the closer to equilibrium you will get.