1.

Abstract

Chromium (VI) which is one of a heavy metal is classified as a strong oxidation agent
and it poses a great deal of toxicity to humans and animals as well due to its
carcinogenetic even at low aqueous solutions concentration. Chromium ions are found
naturally in rivers, lakes, and streams.

This health-risky situation has led to many

studies and investigations on determining the presence and quantity of chromium in

biological and environmental samples. In the present experiment, an absorption
spectroscopy method was used for the determination of chromium (VI) concentration
and the method is operated by tracing the presence of chromium (VI). As a result for
this experiment, the concentration of Chromium (VI) in the lake water sample tested
was 75 p.p.m. The experiment is completed and successfully conducted.
1.2

Introduction

1.3

Aims

The objectives of conducting this experiment are :

1)

To determine the Chromium (VI) content which present in simulated lake water sample

using a spectrophotometer.
2)

To demonstrate the proper method of diluting solution to prepare a series of standard

solutions in the range of 5 to 100 parts per million (p.p.m) to be used in the calibration of the
spectrophotometer.
3)

To analyze whether the stimulated lake water sample tested is suitable for drinking water

and agriculture purposes.

1.4

Theory
Chromium is a metal found in natural deposits as ores containing other elements. The

greatest use of chromium is in metal alloys such as stainless steel; protective coatings on metal;
magnetic tapes; and pigments for paints, cement, paper, rubber, composition floor covering and
other materials. Its soluble forms are used in wood preservatives. One source of chromium
contamination in natural water is automobiles. Chromium that is deposited on roads derived
from brake dusts and exhaust of automobile engine can unfavorably permeate into water
streams through flowing rainwater. The most susceptible areas to be contaminated by such
chromium are lakes and water streams which is nearer a congested with automobiles area.
A number of federal environment agencies are doing researches to determine its
presence in many environmental samples. This is done due to the cancer-risk character and
toxicity. Therefore, in this experiment absorption spectroscopy is being used to detect low level
concentrations of Chromium (VI) in a lake water sample.
Colored aqueous solutions have chemical species contents which absorb significant
wavelengths of light. Similarly, heavy metals can also be determined by absorption of
wavelengths of light. In addition, the amount of light absorbed is linearly proportional to the
concentration of the solution metal ions

The amount of light absorbed by the chemical species in the sample is equivalent to the
difference in the amount of light before it enters the sample and after it exits the sample. The
light must be set to a specific wavelength in order to allow the light to be absorbed by the
chemical species. Every chemical species absorbs distinct wavelengths of light. In absorption
spectroscopy, the wavelength of light absorbed by a metal in solution are detected.

In this experiment, standard solutions will be prepared by diluting 300 parts per million of
Chromium (VI) standard solution. Standard solutions need to be prepared from the concentrated
Chromium (VI) standard solution. The solution dilution formula can be used to calculate the
dilute solutions volume:
(M1) (V1) = (M2) (V2)
Quantitative analysis using spectrophotometer is based on Beer-Lambert Law:
A = LC
A= absorbance value
= molar absorbance (L/mol.com)
L = path length of the cuvette in which the sample is contained (cm)
C = concentration of the compound in solution (mol/L)
According to Beer-Lambert Law, the absorbance value depends on the total quantity of the
absorbing compound in the light path through the cuvette. Thus we will get a straight line
passing through the origin (0,0) if we plotted the graph of absorbance versus concentration of
the compound solution. The molar absorbance, , is a constant for a particular substance,
therefore if the concentration of the solution is halved, so is the absorbance value. A compound
with a high molar absorbance is very effective at absorbing light (of the appropriate wavelength),
and hence low concentrations of a compound with a high molar absorbance can be easily
detected.

1.5

Apparatus

Distilled water
Spectrophotometer
10 millilitres square cuvette bottle
Pipette
5 set of 10 milliliters of 5, 20, 30, 60, 100 p.p.m of diluted solution of
Chromium (VI)
5 set of 300 p.p.m of standard Chromium (VI) solution
Lake water sample

Figure 1 : Spectrophotometer

Figure 2 : Spectrophotometer

1.6

Methodology

In this experiment, the apparatus was set up to carry out the experiment to determine
the concentration of chromium (VI) by using absorption spectroscopy. Firstly, the 10
milliliters of square cuvette bottle was filled up with distilled water and placed on the
spectrophotometer slot. A zeroing step was done to ensure that all the data obtained
later was accurate. It followed by preparing 5 series of diluted chromium (VI) solution by
using the diluted method with distilled water with the range concentration of 1 until 100
parts per million (p.p.m) solutions. Thus, for this experiment it used 5 p.p.m, 20 p.p.m,
40 p.p.m, 60 p.p.m, 80 p.p.m and 100 p.p.m solutions which were 300 p.p.m of standard
Chromium (VI) solution of 0.17, 0.67, 1.33, 2.00, 2.67 and 3.33 milliliters. Each series of
diluted solution was poured into square cuvette bottle and placed on the
spectrophotometer slot to check the absorbance value by using the spectrophotometer.
Then, all the data obtained was recorded first. This experiment continued by preparing
10 milliliters of lake water sample and poured it into the square cuvette bottle and
spectrophotometer was used to determine the Chromium (VI) concentration contained
in the water sample collected. Lastly, all the value was analyzed for the observation and
the data obtained were recorded. The graph was plotted to figure out the value of
chromium (VI) concentration.

1.7 Results

Concentration of Chromium

Volume of chromium

(VI) solution (p.p.m)

(VI) solution (mL)

0.3

0.001

20

1.3

0.011

40

2.7

0.014

60

4.0

0.026

80

5.3

0.034

100

6.7

0.043

Figure 1.7.1.1

Absorbance value

Table of concentration of Chromium (VI) solution and their absorbance value

Absorbance of lake water sample = 0.031

Graph of absorbance value versus concentration of Chromium (VI) solution (p.p.m)

0.05
0.05
f(x) = 0x

0.04
0.04
0.03
Absorbance Value

0.03
0.02
0.02
0.01
0.01
0
0

10

20

30

40

50

60

70

80

90 100 110

Concentration of Chromium (VI) solution (p.p.m)

Graph 1.7.1.2

Graph of Absorbance Value versus Concentration of Chromium (VI) in units

of p.p.m

1.8

Calculations

To dilute 20 milliliters standard solution of 5 p.p.m, the solution dilution formula is used in order
to calculate the volume of Chromium (VI) solution.
(M1)(V1) = (M2)(V2)
(300)V1 = (5)(20)
V1 = 0.3 mL

To dilute 20 milliliters standard solution of 20 p.p.m, the solution dilution formula is used in order
to calculate the volume of Chromium (VI) solution.
(M1)(V1) = (M2)(V2)
(300)V1 = (20)(20)
V1 = 1.3 mL

To dilute 20 milliliters standard solution of 40 p.p.m, the solution dilution formula is used in order
to calculate the volume of Chromium (VI) solution.
(M1)(V1) = (M2)(V2)
(300)V1 = (40)(20)
V1 = 2.7 mL

To dilute 20 milliliters standard solution of 60 p.p.m, the solution dilution formula is used in order
to calculate the volume of Chromium (VI) solution.
(M1)(V1) = (M2)(V2)
(300)V1 = (60)(20)
V1 = 4.0 mL

To dilute 20 mililiters standard solution of 80 p.p.m, the solution dilution formula is used in order
to calculate the volume of Chromium (VI) solution.
(M1)(V1) = (M2)(V2)
(300)V1 = (80)(20)
V1 = 5.3 mL

To dilute 20 mililiters standard solution of 100 p.p.m, the solution dilution formula is used in
order to calculate the volume of Chromium (VI) solution.
(M1)(V1) = (M2)(V2)
(300)V1 = (100)(20)
V1 = 6.7 mL

Based on the graph, the value of the gradient is :

m=

y2 y1
x2 x1

0.042 0.034
100 80
= 0.0004

By interpolating graph, the equation of the linear line was given by as :

Y = mX + C
The y-intercept in the graph is 0.001, so, the value of C = 0.001. by substituting the value of ,
the equation is given as :
Y = 0.0004X + 0.001

X=

Y 0.001
0.0004

By substituting the value of Y with the value of the absorbance of water sample, which is 0.031
abs, the equation is given as :

X=

0.031 0.001
0.0004
X = 75

X is equivalent to the concentration of Chromium (VI) contains in the water sample, which is 75
parts per million (p.p.m).

1.9

Discussion

The experiment was conducted to achieve a few of main objective in this experiment
such as to determine the Chromium (VI) content which present in the water sample by
using the spectrometer, to demonstrate the proper method of diluting solution, to
prepare of standard solution in the range 5 to 100 p.p.m and lastly to analyze whether
the water sample is suitable for drinking water and agriculture purpose.
In chemistry, spectrophotometry is the quantitative measurement of the reflection or
transmission properties of a material as a function of wavelength and the water sample.
The water sample that have been used to examine the presence of chromium is
assumed to be contaminant, the source of the water sample is the source of water
sample located near a roadway where road users with automobiles use it every day.
This is due to rivers, lakes or any water streams which are near to congested areas
have the highest risk or most susceptible to contain heavy metals such as chromium
itself and is classified as contaminated water source.
The first step in this experiment is to prepare six standard solution Chromium (VI)
standard solutions using serial method as mentioned in the theory section. The
concentrations for standard 1, standard 2,standard 3, standard 4, standard 5 and
standard 6 is 100 ppm, 80 ppm, 60 ppm, 40 ppm, 20 ppm and 5 ppm respectively.
The absorbance for each six standard solutions and the lake water sample is taken
by using the spectrophotometer. A graph of the absorbance is plotted against the
concentration of standard solutions. Linear regression analysis is performed in order to
determine the linear best fit for the absorbance versus concentration data.
Graph 1.7.1.2 in results show a graph of absorbance versus concentration of six
standard solutions. The line determined from the regression analysis will be in the form
of y = mx + b, where y is the absorbance value and x is the concentration of the
solution. Algebraic substitution of the absorbance value (y) for the unknown metal
solution into the linear regression equation for the line permits the determination of the
concentration (x) of the unknown solution. The value of absorbance of lake water
sample is 0.031. From the graph, the concentration of chromium (IV) in water sample is
apparently calculated to contain 75 parts per million.
Based on the consideration of state and federal regulatory agencies, natural
waters are regarded to be toxic if the concentration of Chromium (VI) is any higher than
0.05 parts per million. Natural waters whose chromium (VI) concentrations exceed 0.05
ppm may not be used for drinking water or for agricultural purposes. After doing some
calculations based on the graph, the concentration of the lake water sample is

determined to be at 75 ppm which is obviously not suitable for drinking water and for
agricultural purposes. The lake water sample tested is then concluded to be unsafe and
unhygienic for drinking water or even agricultural purposes.
However, a few steps might be done erroneously which results in such an
outrageous value of chromium (VI) content. First and foremost, the techniques applied
to measure such a small volume of diluting substance might not be suitable. This might
lead to difficulties when measuring the volume of chromium (VI) needed to dilute the
standard solutions.
Besides that, wrong way of holding the cuvette bottle used to contain the diluted
solutions might also lead to errors. Any fingerprints or smudge imprinted on the outer
surface of the cuvette can become an unfavorably interference for the solutions to
absorb wavelengths of light in the spectrophotometer. Thus, the reading of absorbance
by the instruments may not be accurate.
In addition, the experiment conducted to determine the absorbance value of
diluted solutions that contain chromium (VI) is done only once. Thus, no average value
can be obtained from the results, and so the accuracy of the value taken is not much
convincing.
Others, the square cuvette bottle might not be cleansed properly with distilled
water each time before it is used for another attempt. This will lead to inaccurate
contents of either standard solutions or the chromium solution.

1.10

Conclusion

Based on the experiment, it can be concluded that chromium (VI) are present in the
sample water. The concentration of Chromium (VI) in the lake water sample tested is
calculated to be 75 parts per million. Since it is above the level of standard allowed by
the Malaysian Environmental Quality (Sewage and Industrial Effluents) Regulations of
1979 which is 0.05 ppm thus, the water is concluded as contaminated and does not
suitable for drinking or agricultural purposes.

1.11

Recommendations

1.
Avoid leaving a fingerprints or any kind of smudge on the outer surface which will corrupt
the reading of absorbance value of spectrophotometer.
2.
Ensure the square cuvette bottle clean thoroughly and properly so that the accurate
reading can figure out.
3.
Use a specific wavelength where the component absorbs substantially to achieve the
accurate value.
4.

Ensure there are no air bubbles in the nozzle of the stopper.

5.
Measure the volume of diluted chromium accurately and make sure read the level of
solution by noting the lower level of the meniscus at eye to avoid parallax error.
6.
Ensure the square cuvette bottle is filled up with distilled water first and placed on the
spectrophotometer slot to ensure the zeroing step is done so that all the data obtained later is
accurate.

1.12

Reference/Appendix