Anda di halaman 1dari 7


C.M. Recto Ave., Lapasan, Cagayan de Oro City


(Modified Spectrophotometric Method)
Experiment No. 5

Submittted by:
John Mark C. Saburao
BS Chem- 3H1

February 19, 2016

At the end of this experiment, the students should be able to:
1. Determine the absorbance of KHP standard solutions by spectrophotometric method
2. Obtain a standard calibration curve by plotting absorbance of KHP standard solutions
versus the concentration of O2
3. Determine the equation of the line of the standard calibration curve
4. Establish absorbance-chemical oxygen demand (COD) relationship for KHP standard
5. Determine and analyze the chemical oxygen demand (COD) of the deep well water sample
by method of least squares or linear regression function


Table 1. Absorbances of Blank and KHP Standard Solutions at 650 nm

KHP Standard Solution (ppm O2 or mg O2/L)

Absorbance at 650 nm

Table 2. Absorbance of Deep Well Sample at 350 nm

Table 3. COD of the Deep Well Water Sample

Absorbance at 650 nm

A modified spectrophotometric method developed by Hisakuni, Takeshi, Takeo & Yukio
in 2001 was employed to determine and analyze the chemical oxygen demand (COD) of a deep
well sample obtained from Kauswagan, Cagayan de Oro City. The method uses the instruments
DR 5000 HACH UV-Vis spectrophotometer and the DRB 200 HACH digester.
Water solutions of standard potassium hydrogen phthalate (KHP) were used as COD
known samples. The concentrations of the KHP solutions were 25, 50, 75, 100, 125, 150, and
200 mg O2/ L or ppm O2. The standard KHP solutions were used to obtain a standard calibration
curve from which the regression equation was obtained and will be used to calculate the COD of
the deep well sample. The KHP is oxidized to carbon dioxide and water as shown in the
chemical reaction below:
C8H4O4- + 7.5 O2

8 CO2 + 3 H2O

The method used in the experiment involved a two hour digestion at high heat (150oC)
under acidic conditions in which potassium dichromate acts as oxidant for any organic matter
present in the water sample. The digester used in the digestion process in order to aid the
complete reduction-oxidation reaction of the solutions was the Digital Reactor Block (DRB) 200
HACH digester. The amount of O2 that is chemically equivalent to the dichromate consumed is
defined as the chemical oxygen demand (COD) of the sample. During the oxidation in which the
sample is heated with the dichromate, organic matter is converted to carbon dioxide and water
while dichromate is reduced to Cr3+:
Cr2O72- + 14 H+ + 6 e-

2 Cr3+ + 7 H2O

The COD results are expressed as the amount of oxygen (in mg O2/L) necessary to carry out the
oxidation of the sample to CO2 and H2O.
Silver sulfate, Ag2SO4, was added to serve as the catalyst and mercuric sulfate, HgSO4,
acts to complex out any interfering chloride. The presence of chloride (Cl-) in the sample is
quantitatively oxidized to Cl2 by the dichromate which interferes with the COD procedure:
Cl- + Cr2O72- + 14 H+ + 6 e-

3 Cl2 + 2 Cr3+ + 7 H2O

The addition of HgSO4 to the reaction mixture complexes with the Cl- and produces a soluble
Hg(II) chloride complex:
Hg2+ + 2 Cl-


Following the digestion, the extent of oxidation is measured through indirect

measurement of oxygen demand via electrons consumed in the reduction of Cr6+ to Cr3+. This
can be done by spectrophotometric methods since potassium dichromate is a colored material
(orange) and is reduced to chromic ion (Cr3+), producing a green solution. The chromic ion
produced simulates the reaction of oxygen with the organic matter in the water sample.
The equation for the calibration curve of the standard KHP solutions or the COD known
samples was determined by employing the method of least squares or linear regression. The

equation of the line was determined to be y = -0.01598 + 9.038x10-3. The graph has a linear
correlation coefficient equal to -0.9987 which indicates there is a strong and direct negative
linear relationship between the absorbance of the known samples and their COD. The negative
value of the correlation coefficient means that as the concentration of oxygen (mg O2/L) in the
known samples increases, the absorbance decreases, hence the graph.
The results for the analysis of the deep well sample showed that it has an average
absorbance of -0.143 at 350 nm wavelength. The oxygen concentration (mg O2/L) or COD of the
sample was calculated using the obtained regression equation. As calculated, the COD of the
deep well sample was 9.51 mg O2/L or 9.51 ppm O2.