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Po
P
= bc
T = Transmittance
A = Absorbance
P0 = Incident radiant power
P = Transmitted radiant power
= Molar absorptivity
b = path length of sample
c = concentration of absorber
Transmittance and absorbance, as defined cannot normally be measured in
laboratory because the analyte solution must be held in a transparent container,
or cell. An experimental transmittance and absorbance that closely approximate
the true transmittance and absorbance are then obtained with the equations:
T = Psolution
Psolvent
A = log Psolution
Psolvent
RESULT
Concentration
(M)
25x10-6
Caffeine
M1V1 = M2V2
M1V1 = M2V2
50x10-6
(25x10-3)(V1) = (25x10-6)
(50mL)
V1 = 0.5mL
M1V1 = M2V2
(25x10-3)(V1) = (25x10-6)
(50mL)
V1 = 0.5mL
M1V1 = M2V2
75x10-6
(25x10-3)(V1) = (50x10-6)
(50mL)
V1 = 1.0mL
M1V1 = M2V2
(25x10-3)(V1) = (50x10-6)
(50mL)
V1 = 1.0mL
M1V1 = M2V2
100x10-6
(25x10-3)(V1) = (75x10-6)
(50mL)
V1 = 1.5mL
M1V1 = M2V2
(25x10-3)(V1) = (75x10-6)
(50mL)
V1 = 1.5mL
M1V1 = M2V2
125x10-6
(25x10-3)(V1) = (100x10-6)
(50mL)
V1 = 2.0mL
M1V1 = M2V2
(25x10-3)(V1) = (100x10-6)
(50mL)
V1 = 2.0mL
M1V1 = M2V2
(25x10-3)(V1) = (125x10-6)
(50mL)
V1 = 2.5mL
(25x10-3)(V1) = (125x10-6)
(50mL)
V1 = 2.5mL
Wavelength
(mn)
212
279
272
272
281
Absorbance
Transition
0.030
0.135
0.405
1.301
1.487
n *
n *
n *
n *
n *
Wavelength
(mn)
224
298
298
278
276
Absorbance
Transition
0.107
0.123
0.093
0.211
0.297
n *
n *
n *
n *
n *
Molar absorptivity
Caffeine
Concentration : 25x10-6
A = lc
0.03 = ()(1 x 25x10-6)
=1200
Concentration: 50x10-6
A = lc
0.135 = ()(1 x 50x10-6)
= 12700
Concentration: 50x10-6
A = lc
0.123 = ()(1 x 50x10-6)
= 2460
Concentration : 75x10-6
A = lc
0.405 = ()(1 x 75x10-6)
= 5400
Concentration : 75x10-6
A = lc
0.093 = ()(1 x 75x10-6)
= 1240
Concentration : 100x10-6
A = lc
1.301 = ()(1 x 100x10-6)
= 13010
Concentration : 100x10-6
A = lc
0.211 = ()(1 x 100x10-6)
= 2110
Concentration : 125x10-6
A = lc
Concentration : 125x10-6
A = lc
Wavelength (mn)
249.50
250
250
Absorbance
25
50
75
100
125
Concentration (ppm)
0.03
0.135
0.405
1.301
1.487
Absorbance
0.057
0.044
0.041
Concentration (x)
y = 0.0163x 0.5524
0.057 = 0.0163(x) 0.5524
x = 37.387 M
Sample 2
A = 0.044
y = 0.0163x 0.5524
0.044 = 0.0163(x) 0.5524
x = 36.589 M
Sample 3
A = 0.041
y = 0.0163x 0.5524
0.041 = 0.0163(x) 0.5524
x = 36.405 M
Average concentration:
= 37.387 M + 36.589 M + 36.405 M
3
= 36.794
Convert to ppm
36.794 mol/L x 194.19g/mol
= 7144.03 g/L = 7.14403mg/L =7.14403ppm
Standard deviation :
1
( Xmean )
N 1
( 37.38736.794 )2
]
1
= 0.522
Relative standard deviation :
= Standard deviation x 100
mean
= 0.522 x 100
36.794
= 1.42%
Actual caffeine concentration in analgesic tablet : 7.14403ppm x 25
= 178.63ppm
Amount of caffeine in sample :
178.63 mg x 0.05 L = 8.93 mg
L
Absorbance
25
50
75
100
125
Concentration
(ppm)
0.107
0.123
0.092
0.211
0.297
Sample
Sample 1
A=0.057
Concentration (x)
y = 0.0019x + 0.0256
0.057 = 0.0019(x) + 0.0256
x = 16.526 M
Sample 2
A = 0.044
y = 0.0019x + 0.0256
0.044 = 0.0019(x) + 0.0256
x = 9.684 M
Sample 3
A = 0.041
y = 0.0019x + 0.0256
0.041 = 0.0019(x) + 0.0256
x = 8.105 M
Average concentration :
= 16.526 M +9.684 M +8.105 M
3
= 11.438 M
Convert to ppm
11.438 mol/L x 180.157 g/mol
= 2060.64 g/L = 2.06064 mg/L =2.06064 ppm
Standard deviation :
1
( Xmean)
N 1
( 16.52611.438 )2
]
1
= 4.483
= 39.2%
Actual acetyl salicylic acid concentration in analgesic tablet :
2.06064ppm x 25 = 51.52 ppm
DISCUSSION
In this experiment, there are two components caffeine and acetylsalicylic
acid that will be determined in the analgesic tablet . Firstly, we need to prepare
five standard s solution of caffeine and acetyl salicylic acid in a different
concentration which are 25ppm, 50 ppm, 75ppm, 100ppm and 125ppm.. Then,
the sample used for this experiment is analgesic tablet that was prepared in
three sample that contained an unknown concentration. All the sample and
standard were run in UV-Vis spectrometer. We will first determine the equation
of each component by constructing a calibration curve absorbance vs
concentration with standard solutions. The maximum wavelength of caffeine
and acetyl salicylic acid are at 270nm and 225nm. The value absorbance in both
are selected based on the wavelength range of UV region.
This combination product contains three medications which are
acetylsalicylic acid (ASA), codeine, and caffeine. ASA belongs to the group of
medications called analgesics (pain relievers), anti-inflammatories, and
antipyretics (fever reducers).Codeine belongs to the group of medications called
narcotic analgesics .Caffeine belongs to the group of medications called
stimulants. ASA - caffeine - codeine is used for the relief of mild-to-moderate
pain, fever, and inflammation. It may be used to treat conditions such as
headaches, pain due to cold symptoms, toothache, pain of menstrual cramps,
arthritis pain, and the pain caused by muscle strains and sprains.
In diagram 1 and diagram 2 shows the graph of absorbance vs
concentration with standard solution for both caffeine and acetylsalicylic acid in
analgesic tablet. The value of R2 in graph diagram 1 is 0.9088 shows a good
precision because the value is near to 1. For diagram 2 the value of R2 is 0.7303
shows the graph is not good precision. This is because the absorbance is not
constantly increase as the concentration increase. From the Beers Law plot, the
equation of straight line that get from diagram 1 and 2 are:
y = mx + c
Diagram 1 : y = 0.0163x 0.5524
Diagram 2 : y = 0.0019x + 0.0256
We will calculated the amount of concentration of caffeine and acetylsalicylic
acid in each of analgesic sample. After we get all the equation for this both
components, the value of the unknown concentration for analgesic tablet will be
determined. From that we can get the actual concentration of sample in average,
for caffeine is 178.63ppm. By using this value, the amount of caffeine can be
calculated which is 8.93mg and the value of the theory should be 15mg. Besides
that, we also calculated the value of standard deviation and relative standard
deviation which are 0.522 and 1.42%. The standard deviation was calculated
because to determine the measure value that dispersed from the average value.
Next, we also calculated the percentage of caffeine which is 8.93%. For diagram
2 which is acetylsalicylic acid, the value of actual concentration of sample in
average is 51.52%. By using this value, the amount of acetylsalicylic can be
calculated which is 2.58 mg and the value of the theory should be 325mg. It
show the value of the result is quite far from the theory. The standard deviation
and the relative standard deviation are 4.483 and 39.2%. The value of
percentage of acetylsalicylic acid is 2.58%.
There will be some errors during do this experiment. Firstly, when
prepared the stock solution some of the caffeine or acetylsalicylic not diluted
enough. This error will effect the concentration of the standard solution.
Therefore, we need to do properly while doing the dilution process. Then, we
need to make sure while preparing the solution there will not have any
contamination occur. Besides, when we use the UV-Vis spectrometer, make sure
the wall of cuvette we used in a clear. If there have any fingerprint on the wall
of cuvette, this will prevent the light beam to penetrate into the solution and
cause the beam to be scattered. So, to prevent this, make sure we clean the clear
wall with tissue or else.
CONCLUSION
In conclusion, we will able to know the maximum absorbance of each
concentration of standard caffeine solution and acetylsalicylic acid solution.
From the graph that had been plotted in diagram 1 and diagram 2, we also can
identify the amount of caffeine and acetylsalicylic acid in analgesic tablet which
are 8.93mg and 2.58mg.
REFERENCES
1) Skoog D.A., Holler F.J., & Crouch S.R. (2007) Principles of Instrumental
Analysis. An Introduction to Ultraviolet-Visible Molecular Absorption
Spectrometry.(6th ed.), United States : David Harris.
2) Acetylsalicylic Acid, Caffeine & 8 mg Codeine Phosphate (n.d),
Retrieved May 14, 2016, from
http://chealth.canoe.com/Drug/GetDrug/Acetylsalicylic-Acid-Caffeine8mg-Codeine-Phosphate
3) The Simultaneous determination of Caffeine and Acetylsalicyclic Acid in
an Analgesic by Ultraviolet Spectrophotometer (n.d.), Retrieved May 14,
2016, from https://www.scribd.com/doc/307257040/EXP-7Simultaneous-determination-of-caffeine-and-acetylsalicylic-acid-in-anAnalgesic-by-ultraviolet-spectroscopy