Quantitative Inorganic Analysis Laboratory

Formal Report
Date Performed: 05
November 2015
Date Submitted: 22
November 2015
Institution/Department:
Department of
Chemical Engineering
College: Engineering
Instructor: Cris Angelo
Pagtalunan

Spectrophoto
metric
Determinatio
n of the Acid
Dissociation
Constant of
Methyl Red

M. K. Coo
Knowing the pKa of an indicator makes choosing one for a titration easier since it
gives the effective range of that indicator. The purpose of the experiment was to
determine the pKa of methyl red (HMR). Multicomponent spectrophotometry was
used to measure the equilibrium concentrations HMR and

.
MR

In this method, a

UV-Vis spectrophotometer was used to measure the absorbance of the HMR and

MR

system and by applying Beers law and systems of equations, the

concentrations of HMR and

MR

were calculated. With these values, the pH of

the solutions, and the Henderson-Hasselbalch Equation, the pKa was calculated.
The experiment yielded measurements of a pKa of 4.45. These values differ from
the literature value of pKa (5.00) by 11%. Two possible sources of error is stray light
and instrument error.

Spectrophotometric Determination of the Acid Dissociation Constant of Methyl Red

Coo, 2015 | 1

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Introduction
In the dissociation of weak acids, the
reaction continues until the concentrations of the
reacting species no longer changes. At this point,
equilibrium is reached which is when the rate of
dissociation is equal to the rate of formation of the acid.
The relation of the concentrations of the reacting species
at equilibrium is related by the equilibrium constant.
Methyl red is a weak acid indicator that becomes
red at acidic pH and yellow at basic pH. The
dissociation reaction of methyl red is shown by
Equation 1.

MR( aq)
H +
( aq ) +
(1)
HMR(aq) ( )
By finding the equilibrium concentrations of the
reacting species of methyl red, the equilibrium
constant can be found. The equilibrium constant
equation of the reaction is shown by Equation 2. 1

MR

+
(2) H

K=

Taking the negative logarithm of both sides, the pKa of

methyl red can be found. pKa 1 gives the range of

pH that methyl red can be used as an indicator. pKa can
be found using linear regression using Equation 3,
which is the Henderson-Hasselbalch Equation. 2 The yintercept of the graph would be the pKa. The pH of the
analyte can be measured using a pH meter.

MR

(3)

pH= pKa+ log

In order to find the equilibrium concentrations of

HMR and

MR , multicomponent spectrophotometry

was used. Spectrophotometry is the use of light to

measure concentration. In this experiment, the

Chem 31.1 Section, Room No., Time

instrument measures the absorbance of the sample of

light of a particular wavelength. Beers Law states that
the absorbance of a species is directly proportional to its
concentration and the path length through the species.
The relationship is shown by Equation 4. 2

Absorbance=abc

(4)

In Equation 4, b is the path length, c is the

concentration of the species, a is the absorptivity
constant. This constant must have a unit that will cancel
the units of path length and concentration and is usually
1/Mcm (the constant would be called the molar
absorptivity constant). 3
For multicomponent analytes, the total absorbance is
equal to the absorbance of all of the species in the
system. Equation 3 can then be modified to take into
account the concentrations of the components. This is
shown by Equation 5. 4
(5)

Absorbance=a1 b c 1+ a2 b c2 +a 3 b c3 + +a n b cn

In Equation 5, there are n components that contribute

to the absorbance,

an

one component and

c n is its concentration. The path

is the absorptivity constant of

length remains constant since these components are

contained in one container. In this experiment, there are
two species that contribute to the absorbance, the redcolored HMR and the yellow-colored

MR .3

The goal of this experiment was to calculate the pKa

and Ka of methyl red using spectrophotometry to
measure the equilibrium concentrations of the
equilibrium reaction between the two species.
Materials and Methods
Two 5 mL portions of the methyl red standard
was placed in two 50-mL volumetric flasks with either 5
mL 0.1 M HCl or 12.5 mL of 0.04 M sodium acetate
and then diluted to mark and mixed thoroughly. The
solution containing HCl and methyl red was labeled as
Solution HMR and the solution with sodium acetate and
methyl red was labeled as Solution MR.
Solutions 1, 2, 3, 4, 5, 6 were prepared in volumetric
flasks and solutions 7, 8, 9, 10 in beakers. The
components of each solution are shown in the appendix.

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The instrument used in this experiment as a doublebeam, UV-Vis spectrophotometer. A double-beam
spectrophotometer has several important parts. A source
produces radiation of many wavelengths. A
monochromator will then disperse the light produced by
the source to a selected wavelength. A chopper will
redirect the path of the light to hit either the cuvette with
the sample or another similar cuvette with the blank
which will absorb an amount of light of the selected
wavelength and allow some of the light to be
transmitted. The detector measures the intensity of the
transmitted light by converting it into an electrical
signal which the computer will convert into data. 3 5
This type of spectrophotometer uses the relation of
absorbance and intensity. The relation is shown by
Equation 5.
(5)

Absorbance=log

I
Io

solutions or the blank solutions as even the best made

cuvettes differ slightly from another. 6
The absorbance values at those wavelengths and pH
of Solutions 7, 8, 9, 10 were measured to calculate the
equilibrium concentrations of HMR and

+
H , respectively.

Results and Discussion

In this experiment, the aim was to calculate
the pKa using multicomponent spectrophotometry. The
measurement of pH is important because this gives the
concentration of

The measured intensity of the blank solution is

MR , and

+
H which is part of the equilibrium

I o to take into account the absorbance of

reaction.
The pH of the solution for weak acids should be at

the cuvette and the solvent, and the intensity of the

of the pKa of a monoprotic acid. A solution

recorded as

I . Then, the computer having a pH of pKa-3 should contain only the acid form

sample is recorded as
calculates the absorbance. 2

The wavelength of maximum absorption,

HMR ,

of HMR was measured by obtaining an absorption

spectrum using Solution HMR and water in the blank
solution. The wavelength corresponding to the peak of

HMR . Similarly, the wavelength of

MR , of
maximum absorption,
was
MR
the graph is

measured by obtaining an absorption spectrum using

Solution MR and water in the blank solution.
In order to obtain the absorptivity constants of HMR
and

MR

at

HMR

least 3

and,

produce only a very small amount of

MR .

Similarly, the pH of Solution MR should be around 8 so

that the dissociation of HMR will be sufficiently
complete so that the amount of HMR remaining is very
small. These measures make Solution HMR and MR
useful for obtaining the wavelengths of maximum
absorption of HMR and

MR .3

Because of the amount of HCl added to the HMR

MR , the absorbance
solution to make Solutions 1,2, and 3, the diluted

of Solutions 1, 2, 3, 4, 5, and 6 were measured at these

wavelengths. From the resulting absorbance vs
concentration graphs, the value of ab (

molar absorptivity constant pathlen gth )

can

be calculated. This is the reason why the cuvettes should

be matched, i.e. the same cuvette is used for all sample

Chem 31.1 Section, Room No., Time

and a solution having a pH of at least pKa+3 should

contain the anion form.7 The theoretical Ka of methyl
red is 5 so the pH of Solution HMR should be around 2
in order to ensure that any dissociation of the HMR will

concentration of the standard methyl red solution is

equal to the concentration of the methyl red (HMR) in
the solution. Using this and the absorbance values at

HMR and

MR , the molar absorptivity constant

can be calculated using linear regression. Since the path

length is set at 1 cm, the slope of the graph is equal to
the molar absorptivity constant. The graph is shown by
Figure 1.
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Linear regression using Equation 3 can also be used

to compute
calculating the pKa. By taking the pH as
/([forHMR])
y and
as x, the y-intercept of the
log
graph corresponds to the pKa. The graph is shown by
Figure 3.
Figure 3

Figure 1.
0.25
0.2

f(x) = 1253.57x + 0.03

R = 0.92

0.15
0.1

6.5

0.05
0
0

f(x) = 71.43x + 0
0 R =00.64 0
0

the absorbance values at

MR , the

molar absorptivity constant can be calculated using

linear regression. The graph is shown by Figure 2.
Figure 2.
0.1
0.08

f(x) = 475x + 0.01

R = 0.9

0.06
0.04
0.02
0
0

f(x) = 78.57x + 0
0 R =00.85 0
0

.
The values of the molar absorptivity constants
calculated is shown in the appendix. Using the values
and absorbance readings of Solutions 7, 8, 9, and 10 at

HMR and

MR , Equation 3, can be modified to

form a system of equations that can be used to calculate

for the concentrations of HMR and

MR . This

system is in the appendix.

Also, using the measured pH and calculated
equilibrium concentrations, pKa can be calculated using
Equation 3.

Chem 31.1 Section, Room No., Time

Linear (YValues)

4.5

MR . Using this and

HMR and

Y-Values

5.5
5

Similarly, because of the amount of sodium acetate (a

basic salt) added to Solution 4, 5, and 6, the diluted
concentration of the standard methyl red solution is
equal to the concentration of

f(x) = 0.22x + 5.44

R = 0.76

From Figure 3, the computed pKa value was 4.45. This

differs from the literature value of 5.0 by 11%. The
relationship between pKa and Ka is logarithmic, which
means that even a small error in the pKa translates to a
much larger error in the Ka.
The average of the pKa values from Table 2 varies
by 11%. These pKa values, computed from Table 3, has
a standard deviation of 0.0002. This means that the
computed values do not stray far from the mean. This
indicates a precise experiment.
A possible source of error was the blank solution for
the experiment. This only contained water, whereas the
solution contained either HCl, sodium acetate, acetic
acid or a mixture of these reagents. This may cause the
measured absorbance to include the absorbance of these
species. However, the fact that these species are not
colored reduces the effect of this error. Also, the
container may be contaminated by interfering
substances which changes the absorption of the solution
by scattering the light or changing the refractive index
of the solvent. 2 3
Also, the cuvette is a possible source of error. The
transition from air to cuvette to solution may cause
refraction which may cause the measured absorbance to
change. Stray light is also a source of error which
causes the measured absorbance to decrease. 4
Instrument error is also a possible source of error.
The monochromator may not be able to completely
eliminate light of other wavelengths and since Beers
law assumes monochromatic light, this can lead to

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changes in absorbance readings. This error is reduced by
selecting the

max

of both species.7

Also, the absorbance reading at

HMR

for

Solutions 5 and 6 was displayed as the same. However,

the concentration of methyl red in Solution 6 is less than
the concentration of methyl red in Solution 5. This
means that the absorbance of Solution 6 should be
lower. The very low absorbance readings of Solution 5
and 6 indicates that they are too dilute to make accurate
readings. This causes the molar absorptivity constant of

MR

at

HMR to change since the graph is no

longer linear. To minimize this error, the concentrations

can be increased but not by too much since this can also
cause an error due to a limitation of Beers Law. This
limitation limits the concentration to a maximum of 0.1
M. 3
Conclusions and Recommendations
The goal of the experiment was to determine
the pKa of methyl red. To measure the pKa,
multicomponent spectrophotometry was used to
measure the concentration of the acid and anion form
of methyl red and a pH meter to measure the

+
H

concentration. This allows the use of the HendersonHasselbalch Equation to calculate pKa and pH. The
pKa calculated was

4.45 0.0003 .

To improve accuracy, it is recommended that the

experiment be done with more sample solutions. This

makes the pH vs

MR

log

Chem 31.1 Section, Room No., Time

accurate by adding points to the graph. Also, it is

recommended that the cuvettes be selected such that it
does not refract light much and that there are minimal
differences in the dimensions of the cuvettes. It is also
recommended that the concentrations be increased to
increase the absorbance and that the blank contain the
added reagents in the solutions and not just water
References
[1] Colby College. pKa of Methyl Red.
http://www.colby.edu/chemistry
/PChem/lab/pKaMethylRed.pdf (accessed November
16, 2014)
[2] Harris, D.C. Quantitative Chemical Analysis, 8th
ed.; W.H. Freeman & Company: New York, 2010.
[3] Skoog, D. A., West, D. M., Holler, F. J., Crouch, S.
R. Fundamentals of Analytical Chemistry, 9 th ed.;
Brooks/Cole: Belmont, 2014.
[4] Harvey, D. Modern Analytical Chemistry; McGrawHill: New York, 2000.
[5] Silberberg, M. Chemistry: The Molecular Nature of
Matter and Change, 5th ed.; McGraw-Hill: New York,
2010.
[6] Jeffery, C.H., Bassett, J., Mendham, J., Denney, R.C.
Vogels Textbook of Quantitative Chemical Analysis,
5th ed.; John Wiley & Sons, 1989.
[7] Patnaik, P. Deans Analytical Chemistry Handbook,
2nd ed.; McGraw-Hill: New York, 2004.

graph to become more

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