SPECTROPHOMETRIC DETERMINATION OF PH OF WATER SAMPLE

Gobe jam chem 301

INTRODUCTION
The pH of an unknown solution is calculated using the spectrophotometric approach by adding a
small amount of a pH indicator and measuring the degree to which the indicator dissociates (a
weak acid) , Byrne, R.H., 1985). Measurement of the absorption of photons in a certain spectral
band is known as spectrophotometry. A spectrophotometer consists of spectrometer for creating
light of any desired wavelength and a photometer for measuring light intensity (number of
photons). In this experiment, the amount of light absorbed in relation to wavelength is measured
using spectrophotometry.The Beer-Lambert law or Beer’s law states that the quantity of
attenuation relies on the concentration of the absorbing molecules and the path length which
absorption occurs. According to this formula, absorbance is directly proportional to the
concentration of the solution (c), and to the path length (b), which is equal to the length of the
cuvette. Acid-base indicators are used to show the endpoints of acid-base reactions. Most acid-
base indicators, including bromocresol green, are regarded as weak acids. The indicator is a
complex molecule that can be represented with the formula Hln, where ln- represents the anion.
When applied to low concentrations, the indicators signal pH changes within a particular range,
which is variable among the indicators,(Cantrell, K.J., 2013). The color change depends on the
relative acid strength (pKa) of the conjugate acid form of the indicator.
Acid/base indicators are weak acids or bases. When dissolved in water they dissociate and form
ions. Consider an acid type indicator, HIn. Its behavior can be described by the equilibrium
equation.
HIn + H2O In- + H3O+

Quantitatively, the relationship between the hydronium concentration, H3O+, and the relative
amounts of Hln and ln– in solution is described by the acid-dissociation constant, Ka. Based on
the equation above, the acid strength is calculated as follows

and

Aside from using a pH meter, Ka can also be determined spectrophotometrically by employing

Beer Lambert’s law.

OBJECTIVE
 This experiment aims to determine the pKa of an acid-base indicator, Bromocresol
Green, using spectrophotometer.
MATERIALS
 1. REGEANTS
 Bromocresol green
 HCl
 NaOH

2. EQUIPMENT
 Volumetric flask
 Graduated cylinder.
 Water
PROCEDURES
Determination of Individual Absorption Spectra
25.00 ml aliquot of the bromocresol green indicator was transferred to two 100 ml volumetric
flask and to one of them 25 ml of 0.5 M HCl was added and to the other flask 25 ml of 0.4 M
NaOH was added. Then water was adder to both flasks to make dilution to the mark and mixed
to make solution homogenous. The absorption spectra for the acids and conjugate were then
obtained.
Determination of pH of an Absorption Spectra
25 ml of stock bromocresol indicator green was transferred in the 10 ml volumetric flask
containing 50.0 ml of the of the unknow solution then it was diluted to the mark with water then
mixed well. Measure the absorbance of the diluted solution at the wavelength for which
absorptivity data were calculated.

RESULTS AND DISCUSSION

Absorbance measurements were performed using a Spectrophotometer in the wavelength range
between 400 and 650 nm. Plots of absorbance against λ for the bromocresol green indicator in
buffers with different pH levels are illustrated in Figures 1, 2, and 3
Figure 1 ; Sample of NaOH+ BC

GRAPH OF ABSORBANCE VS WAVELENGTH

4

3.5

3
ABSORBANCE ,nm

2.5

1.5

0.5

0
350 400 450 500 550 600 650 700
WAVELENGTH

Figure 2; Sample of HCl

Figure 3; Sample of Unknown solution
Table 1: Absorbance values at 500nm, 590nm and 650 nm wavelengths as a function of pH were
selected to determine the pH.
NaOH HCL nm
1.097 0.787 500
3.149 0.035 590
1.970 0.020 650

CALCULATION
Finding molar absorbtivities
HCl =0.5 M
NaOH = 0.4 M
Molar absorptivity for NaOH

1. A= ɛ b C
1.097 = ɛ× 1 cm× 0.4M
ɛ = 1.097/ 0.4M = 0.439

2. A = ɛ × b ×C
3.149 =ɛ ×1cm ×0.4
ɛ =3.149/ 0.4
= 7.87

Molar absorptivity for HCl

1. A= ɛ× b ×C
0.787 = ɛ×1cm ×0.5
ɛ = 0.787 / 0.53
= 1.484

2. A= ɛ ×b× C
0.035 = ɛ× 1cm × 0.5
ɛ = 0.035/0.5
 = 0.07

pKa= -log Ka
= -log (1.6×10-5)
=4.795880017
pKa = 4.8
pH= pKa + log K
=4.8+ log (0.33)
=4.32
The pKa value is essential and unique to every indicator. In this experiment, two methods were
employed to determine pKa (Byrne, R.H. and Yuan, D., 2019). The first method revolved around
measuring the pH of the buffer solutions and determination of the absorbance. From the results,
as in Figure 4, the inflection point was obtained, and it was found to be 4.8 (inflection point =
pKa). As a result, the experimental value of this pKa was almost equal to the theoretical pKa of
bromocresol green that is 4.7. The percentage error can therefore be calculated as: % error =
experimental value – actual value / actual value x 100% = 4.5 – 4.7/4.8 x 100% = 97.9%. The
experiment had an experimental error of 4.4% which is small, thus acceptable. The second
method, on the other hand, used the Beer's Additive Law and the Henderson-Hassel bach
equation to ascertain the linear relationship between pH and solution concentration. The pKa for
the second approach was calculated to be 4.46, which is roughly the same as the pKa for the first
method. The discrepancy between the theoretical and observed pKa values points to possible
experimental inaccuracy. This could be the result of erroneous measurements of the volume of
solutions or stray radiation that interfered with the absorbance and ionic strengths of the various
solutions. Cantrell, K.J., 2013.

CONCLUSION
The application of spectrophotometric titration allowed the pKa of Bromocresol Green to be
determined. Two methods of analysis (graphical and additivity of Beer’s Law) were employed
and generated results with good similarity. The pka was found to be 4.8 And the pH was found to
be 4.3.
REFERENCE

 Ma, J., Shu, H., Yang, B., Byrne, R.H. and Yuan, D., 2019. Spectrophotometric
determination of pH and carbonate ion concentrations in seawater: Choices, constraints
and consequences. Analytica Chimica Acta, 1081, pp.18-31.
 Shao, H., Thompson, C.J., Qafoku, O. and Cantrell, K.J., 2013. In situ
spectrophotometric determination of pH under geologic CO2 sequestration conditions:
method development and application. Environmental science & technology, 47(1), pp.63-
70.
 Yao, W. and Byrne, R.H., 2001. Spectrophotometric determination of freshwater pH
using bromocresol purple and phenol red. Environmental science & technology, 35(6),
pp.1197-1201.

 Robert-Baldo, G.L., Morris, M.J. and Byrne, R.H., 1985. Spectrophotometric

determination of seawater pH using phenol red. Analytical Chemistry, 57(13), pp.2564-
2567.