Professional Documents
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Criteria Page
Index 1
Objectives 2
Summary 2
Introduction 3-7
Materials and Apparatus 8
Procedures 9-10
Results 11-14
Discussion 15-19
Conclusion 19
Recommendation 20
Tutorial 21-22
References 23
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OBJECTIVE
To determine the pKa of methyl red by using the absorbance spectra as a function of pH.
SUMMARY
Methyl red (MR) was pipette into a volumetric flask. Ethanol (95%) was added and was diluted
with distilled water.
MR standard solution was pipette into volumetric flask and HCl (0.1M) was added. Distilled
water was used to dilute the solution. The pH value of the solution was checked and the colour of
the solution was observed. The absorption spectrum for the solution was recorded.
MR standard solution was pipette into volumetric flask and NaOH (1.0M) was added. Distilled
water was used to dilute the solution. The pH value of the solution was checked and the colour of
the solution was observed. The absorption spectrum for the solution was recorded.
Absorption spectrum at the eight different pH values was recorded. The volume of the mix
solution was given in the table 1.0. The quantity of the substance as in table 1.0 was pipette in
volumetric flask and diluted with distilled water. The flask was shook well to homogenize the
solution. The absorption spectrum for each solution was recorded on a different paper. The
remaining solution was kept to determine the true pH value by using the pH meter.
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INTRODUCTION
-1-
[ ][ ]
-2-
[ ]
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The prime indicates that used concentrations rather than activities. Activities are necessary in
true thermodynamic equilibrium constants. Using concentrations, instead, gives the effective or
conditional equilibrium constant.
By definition pH = log [H+] and pKa = log Ka. Taking the ( log) of both sides of equation 2
gives:
[ ]
-3-
[ ]
In this experiment, this equilibrium constant, pKa', will be determine by varying the pH and
[ ]
measuring the ratio . The acetic acid-acetate buffers will be use to control the pH values,
[ ]
since the Ka value for acetic acid is in the same range as the Ka value for methyl red. The pH of
these buffers force methyl red to distribute itself somewhat evenly between the two coloured
forms.
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The absorption spectrum can be symbolized as lambda (). Lambda () max can be obtained at
the highest peak of the absorption spectrum graph.
The values of the [HMR] and [MR-] can be obtained by using the equation given:
-4-
-5-
A = e [X] -6-
where,
A = absorbance
e = molar absorption coefficient
= path length of the cell in centimetres
[X] = concentration of the absorbing species in moles per litre
The absorbance of mixtures is the sum of the separate absorbencies. In mixtures of the acid and
base forms of methyl red the total absorbance is
-7-
The absorption spectra of [HMR] and [MR-] are given schematically in Figure 2. For two
components in solution, the absorbance must be measured at two different wavelengths. The best
wavelengths to choose for the analysis are where one form absorbs strongly and the absorbance
of the other form is negligible. Examination of Figure 2 reveals that there are no wavelengths
where one form, acid or base, absorbs exclusively. For this case, two equations need to be set up
in two unknowns, one equation for each wavelength. Call the two wavelengths l1 and l2. The
absorbance at l1 is A1 and at l2 is A2.The two measurements then provide two simultaneous
equations with two unknowns:
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[ ] [ ] -8-
[ ] [ ] -9-
The molar absorbance coefficients are illustrated in Figure 2. The molar absorbance coefficients
are determined from standard solutions that contain one component alone. Equation 6 and 7
provide two equations in two unknowns. For an unknown solution, the absorbance at the two
wavelengths, A1 and A2, are determined and then equation 6 and 7 are solved for the unknown
concentrations [MR-] and [HMR] at each given pH.
Figure 2.0: Absorbance of a solution is the sum of the absorbencies of the constituents.
Measurements at two wavelengths are necessary to determine the composition of
a two- constituent solution if the absorbance bands overlap. The first subscript
indexes the wavelength and the second subscript indexes the constituent.
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In spectroscopy, an isosbestic point is a specific wavelength at which two chemical species have
the same molar absorptivity () or -more generally- are linearly related. The word derives from
two Greek words: isos: equal and sbestos: extinguishable. [1]
Figure 3.0
Then, the theoretical graph that should be obtained between absorption spectrum against pH
values of the mixtures is like Figure 4.0. Draw a straight line corresponding to the absorption at
pH 1 (Line 1) and pH 13 (Line II). Hence, the graph obtained as in Figure 4.0.
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MATERIALS
Methyl Red
Ethanol (95%)
Hydrochloric Acid, HCl (0.1M)
Sodium Hydroxide, NaOH (1.0M)
Sodium Acetate (0.2M)
Acetic Acid (0.2M)
APPARATUS
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PROCEDURE
1. 5.0mL of methyl red (MR) stock solution was pipette into a 100mL volumetric flask.
2. 50mL of ethanol (95%) was added and diluted with distilled water to obtain 100mL
solution.
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Part C: Absorption Spectrum of MR at various pH
1. The absorption spectrum was recorded at the eight different pH values; besides the two
pHs in the Part B above.
2. As a guideline, to obtain the specific pH value, the volume of the mix solution was given
in Table 1.0.
3. To prepare a solution with a specific pH, the quantity of the substance was pipette as in
Table 1.0 in 50mL volumetric flask. The solution was diluted with distilled water to the
50mL calibration mark.
4. The flask was shook well to homogenize the solution.
5. The absorption spectrum for each solution was recorded on a different paper. Recording
should be carried out immediately after preparing the solution.
6. The remaining solution was kept to determining the pH value.
7. The true pH value for all the solutions was determined by using the pH meter.
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RESULT
Part B:
Methyl Red Solution pH Values Colour Changes
Acid solution 1.8 Light red to dark red
Basic solution 13.45 Light red to yellow
Table 1.0
Part C:
Volume of Volume of Acetic Volume of Sodium
Solution pH Abs ( max)
MR/mL Acid (0.2M)/mL Acetate (0.2M)/mL
A 6.0 12.0 8.0 4.36 2.469
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Overall results:
2.5
2
Absorption
1.5
Absorption
Linear (Absorption)
1
0.5
0
0 5 10 15
pH values
Graph 1.0
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Absorption,
Wavelength (nm)
pH1 A B C D E F G H pH13
600.0 0.055 0.070 0.063 0.057 0.048 0.037 0.032 0.030 0.028 0.007
575.0 0.536 0.645 0.575 0.503 0.402 0.304 0.246 0.200 0.120 0.009
550.0 1.863 1.472 1.298 1.138 0.899 0.676 0.549 0.433 0.247 0.013
525.0 2.432 1.587 1.402 1.232 0.977 0.743 0.609 0.492 0.296 0.036
500.0 2.149 1.307 1.171 1.044 0.860 0.700 0.605 0.553 0.423 0.144
475.0 1.272 0.850 0.810 0.780 0.729 0.707 0.681 0.749 0.725 0.361
450.0 0.539 0.508 0.541 0.584 0.632 0.714 0.738 0.898 0.952 0.522
425.0 0.018 0.337 0.397 0.465 0.551 0.667 0.712 0.896 0.980 0.549
Table 4.0
2.5
pH1
Isosbestic point A
2
(470, 0.75) B
Absorption
C
1.5
D
E
1
F
G
0.5
H
pH13
0
425 445 465 485 505 525 545 565 585 605
Wavelength (nm)
Graph 2.0
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Use these equations for calculating the pKa, [MR-] and [HMR] values.
[ ]
-3-
[ ]
-4-
-5-
0
3 3.5 4 4.5 5 5.5 6 6.5
-0.2
Log [MR-] / [HMR]
-0.4
Log [MR-] / [HMR]
-0.6 Linear (Log [MR-] / [HMR])
-0.8
y = -0.4654x + 1.8722
R = 0.4684
-1
-1.2
pH values
Graph 3.0
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DISCUSSION
The acid solution of Methyl Red (MR) was prepared at the pH value of 1.8 and the colour
change for this solution is light red to dark red while the basic solution of methyl red obtained
was 13.45 of pH value. The colour of the basic solution turned from light red to yellow. The
absorption spectrums of the solution for both solutions were recorded in the length of 350
600nm. The absorption spectrum of acidic solution will produce high absorption of spectrum
Absorption spectrum is the characteristic pattern of dark lines or bands that occurs
equivalent pattern occurs as coloured lines or bands in the emission spectrum of that medium.[3]
For solution A, 12.0mL of Acetic Acid (0.2M) was mix with 8.0mL of Sodium Acetate
(0.2M) and 6.0mL of Methyl Red solution. The pH obtained by using pH meter was 4.36 and the
maximum absorption of spectrum was 1.587. Hence the higher the volume or concentration of
acid used, the maximum absorption of spectrum will be higher. The pH value of the solution will
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Overall Reactions
Graph 1.0 shows the relationship between the absorption of spectrums against pH values.
Line I represents for the absorption of spectrum at pH 1 and Line II represents for the absorption
of spectrum at pH 13. The theoretical graph that should be obtained is shown below.
The graph that that has been plotted was not similar with the theoretical graph because of several
problems happen while conducting this experiment such as the cuvette was not cleaned properly
and there were finger printing on the cuvette that caused the UV and invisible light cannot
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Graph 2.0 shows the relationship between the absorption of spectrum against wavelength
(nm). The isosbestic point that has been obtained when plotting all the graphs was (470, 0.75)
where 470 and 0.75 represented the wavelength and the absorption of spectrum respectively.
From the graph obtained, the isosbestic point of the overall lines of absorption can be determined
by marked the intersection point of the similar level of absorption of the each pH values. The
levels of the absorption spectrum of pH 1 and pH 13 were not same to the others because of
During a chemical reaction, a point in the absorption spectrum (that is, a wavelength)
where at least two chemical species (for example, reactant and product) have identical molar
absorption coefficients, which remain constant as the reaction proceeds. A stable isosbestic point
[ ]
Graph 3.0 shows the relationship between and pH values. The graph obtained
[ ]
was inversely proportional. The intersection point at X-axis is the point where the indicator
concentration in the acidic medium is the same as the concentration in the basic medium. At the
pH where the intersection occurs, the pKa of the MR indicator will be determined.
[ ]
From each results obtained, that graph 3.0 shown always inversely
[ ]
proportional against pH value. The indicator concentration in the acidic medium must be
equally as the concentration in the basic medium. So, the intersection point of the graph at x-axis
must be taken to get the value of pKa of the methyl red indicator. The pKa values must be same
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with the pH values obtained by using theoretical method. The value of pKa can be determined by
There are few points why the lines of the graphs deviate from the best fit line.
i. The mixtures had been contaminated during the experiment.
ii. The volumes of the mixtures were incorrect due to parallax error during measuring the
mixtures. (The eyes level of observer was not at the same level of the meniscus.)
iii. The solution may vaporize due to the property of strong oxidizing agent. Vaporization
may cause the volume of the solution being reduced without any notice.
iv. Different surrounding temperature. Should be conducted in a closed room with a stable
room temperature.
There were few weaknesses in this experiment. Firstly, there were bubbles in the pipette
appeared when the solutions were pipette. Then, the volume will decrease and the concentration
changed thus affecting the pH values. Furthermore, the apparatus used for the experiment were
not well cleaned. There was lot of impurities or known as foreign substances stacked with the
apparatus that has been used. The contamination of the solution caused error to the result.
Parallax error caused different volume when measuring the solution thus, the pH values obtained
will be wrong. Moreover, the cuvette from the spectrophotometer used was not clean properly
and there was fingerprint on it. So, the UV light that went through the silica cell will be affected.
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There are few precautions and safety steps that need to be taken when conducting this
experiment.
1. Avoid skin contact with acid and basic solution (use gloves when handling the acid).
2. Use stopper to prevent the spilling of the solutions from the volumetric flasks.
CONCLUSION
The objective of this experiment of this experiment is to determine the pKa of monobase
indicator that is methyl red by measuring the absorbance spectra as a function of pH by using
ultraviolet visible spectrophotometer. The values of pKa can be determined by using the
formula given:
[ ]
-3-
[ ]
At this point, the overall lines of absorption can be determined by marked the intersection
point of the similar level of absorption of the each pH values. This point is called the isosbestic
[ ]
From each results obtained, that graph 3.0 shown always inversely
[ ]
proportional against pH value. The indicator concentration in the acidic medium must be
equally as the concentration in the basic medium. So, the intersection point of the graph at x-axis
must be taken to get the value of pKa of the methyl red indicator and the pH value of the solution
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RECOMMENDATION
To run this experiment, there are few recommendations that need to be followed to ensure the
1. Use stopper to close and prevent the impurities from entering the volumetric
flask.
2. Make sure the eye level and the meniscus of the solution and calibration mark are
3. Before use the apparatus, make sure that the apparatus is cleaned perfectly by
4. Clean the pH meter using distilled water before use to measure the pH value.
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QUESTIONS
From each results obtained, it shows that graph log [MR-]/[HMR] against pH values
always in a straight line. The indicator concentration in the acidic medium need to be
same as the concentration in the basic medium hence the graph will have the intersection
point at x axis. Then, from the intersection at the graph, we can get the value of pKa of the
2. From the data, is the pKa value similar with pH? If so, state the condition why it is
The pKa values almost same with the pH values obtained using pH meter. In theoretical
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3. How do you calculate the pH of the buffer solution prepared? (One example is
sufficient). Is the pH value from the calculation are the same as the value determined
using pH meter?
So, the difference is 0.07 which is not so far compared with the value from pH meter.
When comparing the pH value by calculated with the value that obtained from the pH
meter, the values are different because by using formula the value will be fixed. It is
different with pH meter which is not stable at all and depends on how the user use the pH
meter.
Less accuracy measurement of the mixtures. Its known as parallax error which is the
The dilutions of the mixtures are not exact. The meniscus level more than the calibration
The cuvette is not well cleaned. It will cause the cuvette less transparent and interfere the
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REFERENCES
1. Isosbestic Point,
2. Spectrophotometer,
3. Absorption Spectrum,
4. Isosbestic Point,
20:59
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