ABSTRACT

People can be exposed to chromium through breathing, eating or drinking and through
skin contact with chromium or chromium compounds. The level of chromium in air and water is
generally low. In drinking water the level of chromium is usually low as well, but contaminated
well water may contain the dangerous Chromium (IV) or hexavalent chromium. Chromium (VI)
is classified as a strong oxidation agent and it poses a great deal of toxicity to organisms due to
its carcinogenetic where it can alter genetic materials and cause cancer. The main human
activities that increase Chromium (VI) concentrations are chemical, leather and textile
manufacturing, electro painting and other Chromium (VI) applications in the industry. These
applications will mainly increase concentrations of Chromium in water.

This health-risky

situation has lead to many researches and investigations on determining the presence and
quantity of chromium in biological and environmental samples. In the present study, an
absorption method for the determination of Chromium (VI) concentration is proposed and this
method is operated spectrophotometrically by tracing the presence of Chromium (VI) at a
wavelength of 435 nanometres. The concentration of Chromium (VI) in the water sample tested
is determined to be 26.12 parts per million where this value is more than what has been
standardized by state and federal regulatory agencies.

INTRODUCTION
Many heavy metals, such as chromium, are undeniably toxic at low aqueous solution
concentrations. Chromium ions are commonly found in either trivalent, Chromium (III), or
hexavalent, Chromium (VI), states. Chromium (VI) formation is favored by an oxidizing
environment, while Chromium (III) is favored by a reducing environment. It is found that
naturally alkaline rivers and lakes have much lower risk of such heavy metal contamination of
chromium ions in comparison with naturally acidic water streams.
Chromium (VI) is known to be a strong oxidizing agent, which apparently poses a high
risk yet negative impact to humans and animals due to its carcinogenetic properties. That is why
there are lots of studies that have been done profusely in order to determine the health-risky
chromium in environmental as well as biological samples.

OBJECTIVES
The objectives of conducting this experiment are to determine the Chromium (VI)
content which present in simulated lake water sample using a spectrophotometer, to
demonstrate the proper method of diluting solution to prepare a series of standard solutions in
the range of 1 to 100 parts per million (p.p.m) to be used in the calibration of the
spectrophotometer, and thus, to analyze whether the simulated lake water sample tested is
suitable for drinking water and agriculture purposes.

THEORY

The main human activities that increase Chromium (VI) concentrations are chemical, leather
and textile manufacturing, electro painting and other Chromium (VI) applications in the industry.
Automobiles industries is also one of the common source of chromium contamination in natural
waters as it is contain a component in various automobile brakes as well as parts of engine.
Chromium that is deposited on highroads originated from brake dusts and exhaust of
automobile engine can unfavorably permeate into any water streams nearby via flowing
rainwater. Therefore, lakes and water streams which are nearer a typical congested with
automobiles areas are the most susceptible areas to be contaminated by such chromium. Due
to its cancer-risk character and toxicity even at low concentrations, it lead a numerous of federal
environmental agencies to do researches in order to determine the presence of Chromium (VI)
in many natural samples such as lake, river and many more.
The absorption spectroscopy will be used in this experiment to identify low level
concentrations of Chromium (VI) in a lake water sample. By using absorption spectroscopy,
heavy metals can be identified via absorption of wavelengths of light. The amount of light
absorbed is linearly proportional to the concentration of the solution metal ions. Absorption
spectroscopy operates on the measuring principle of light before and after it passes through an
aqueous metal solution. 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. For the purpose of light to be absorbed by chemical species, the light must be set to
a specific wavelength. For every chemical species they will absorbs different wavelengths of
light. Absorption spectroscopy is function to detect the wavelengths of light absorbed by a metal
in solution. In this experiment, five standard solutions will be prepared in differences
concentration which is in range 1 to 100 parts per million of Chromium (VI) by diluting it with
solvent which is distilled water. The volume of dilute solutions can be calculate by using the
solution dilution formula which is:
MiVi = MfVf
Where, M is molarity, V is volume, and the subscripts i and f refer to the initial and final values
(Helmenstine, A. M.).

For each wavelength of light passing through the spectrometer, the intensity (I0) of the light
passing through the reference cell is measured. The intensity (I) of the light passing through the
sample cell is also measured for that wavelength. If I is less than Io, then obviously the sample
has absorbed some of the light (Ingle, J. D et al 1988). The relationship between absorbance
(A) and the two intensities is given by:

The quantitative analysis by using spectrophotometer is based on Beer-Lambert Law which is:
A = LC
Where,
A = absorbance value (dimensionless, thus no units is represented)
= molar absorbance (L / mol.cm)
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 it states that absorbance value is depends on the total quantity
of the absorbing compound in the light path through the cuvette. Thus, if a graph of absorbance
versus concentration of the compound solution is plotted, a straight line passing through the
origin will be obtained. 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 at the appropriate wavelength,
and hence low concentrations of a compound with a high molar absorbance can be easily
detected. As what has been standardized by state and federal regulatory agencies, the water
sample that is more than 0.10 parts per million concentration of Chromium (VI) can be
concluded to be contaminated and thus it is not suitable for drinking or agricultural purposes.

APPARATUS / REAGENTS
-

Distilled water

spectrophotometer

10 millilitres square cuvette bottle

pipette

5 set of 10 millilitres of 20, 40, 60, 80 and 100 p.p.m of diluted solution of Chromium (VI)

5 set of 300 p.p.m of standard Chromium (VI) solution of 6.4, 12.8, 19.23, 25.64 and
32.05 millilitres.

Lake water sample.

PROCEDURE
1. 20mL of the 300.00 ppm Cr (VI) standard solution was obtained.
2. Using the serial dilution method, 10.00 mL each of five standard solutions having
concentration of 1.00 tp 100.00 ppm were prepared. One of the standard solutions must
have a concentration of 1.00 ppm and one must have a concentration of 100.00 ppm.
Students are to choose the concentration of the three remaining standard solutions
(concentration must be within the 1.00 to 100.00 ppm range). 10.00 mL volumetric flask,
pipettes and distilled water is used to prepare the standard solutions.
3. The absorbance spectra of the 5 solutions were recorded and an absorbance spectrum
of the simulated lake water sample using the spectrometer. All chromium solutions were
poured into the Waste Container when the experiment is concluded.
4. The tab delimited files containing the absorbance data for each of the five standards
solutions was opened.

for one of the absorbance bands was selected (this might

be an arbitrary determination as the absorbance fluctuates near the wavelength of the

maximum absorbance). The data was recorded.
5. The absorbance concentration for the Cr (VI) standards (from most to least
concentrated) were entered, and the corresponding absorbance reading

for each

standard solutions were recorded.

6. A Beer-Lambert curve was prepared by plotting the absorbance versus data in the table
in step 5.
7. The absorbance of the water sample at

was substituted into the equation for the

line produced in step 6, and the concentration (x-value) of Cr (VI) in the simulated lake
water sample were calculated.

RESULT

Table 1: Determination of absorbance value using spectrophotometer

Volume of Cromium (IV)

Concentration (ppm)

Absorbance

(mL)

20

6.4

0.237

40

12.8

0.504

60

19.23

0.751

80

25.64

1.017

100

32.05

1.252

Absorbance value for water sample: 0.324

1.4
y = 0.0126x - 0.0051
R = 0.9997

1.2

Absorbance

1
0.8
0.6
0.4
0.2
0
0
-0.2

20

40

60

80

100

120

Concentration (ppm)

Figure 1: Absorbance versus concentration

SAMPLE OF CALCULATION

1.

M1V1 = M2V2
(156 mg/L) V1 = (20mg/L)(50 mL)
V1 = 6.4 mL
20 ppm = 6.4 mL

Mg/L = ppm

For concentration 40, 60, 80 and 100 ppm.

40 ppm = 12.8 mL
60 ppm = 19.23 mL
80 ppm = 25.64 mL
100 ppm = 32.05 mL

2.

From graph, the equation of linear line is given as:

y = 0.0126x 0.0051
x =

Substitute the value of y which is 0.324

x =
x =26.12 ppm

The concentration value of chromium (VI) contain in water sample is 26.12 parts per million.

DISCUSSION

This experiment is conduct to accomplish several objectives which is to determine the

Chromium (VI) content which present in water sample using a spectrophotometer, to
demonstrate the proper method of diluting solution to prepare a series of standard solutions in
the range of 20 to 100 parts per million (p.p.m) to be used in the calibration of the
spectrophotometer, and thus, to analyze whether the water sample tested is suitable for drinking
water and agriculture purposes.
The water sample used to examine for presence of chromium is assumed to be
contaminated as the source of water sample is located near a roadway where road users with
automobiles use it every day. This is due to rivers, lakes and 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 then classified as contaminated water source.
In addition, according to the values of concentration of chromium (VI) which is
apparently calculated to contain 26.12 parts per million. These show that there is a significant
content of chromium ions in the water. 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 0.100 parts per million. The water sample tested is then concluded to be
unsafe and unhygienic for drinking water for public consumption or even agricultural purposes.
On the other hand, a few steps might be done inaccurately such as wrong way of
holding the square 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
unfavourably interference for the solutions to absorb wavelengths of light in the
spectrophotometer. Thus, the reading of absorbance by the instruments may not be accurate.
Besides that, 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 realistic.
Other than that, the square cuvette bottle might not be cleansed properly with distilled
water each time before it is used for another test. This also will lead to inaccurate contents of
either standard solutions or the chromium solution.

CONCLUSION
The objective of doing this experiment is to determine the concentration of Chromium
(VI) via absorption was achieved. From the results obtained, the concentration of Chromium (VI)
in the water sample tested is 26.12 parts per million. Since it is more than 0.10 parts per million
as what has been standardized by state and federal regulatory agencies, the water sample can
be concluded to be contaminated and thus it is not suitable for drinking or agricultural purposes.
This is due to the high concentration of Chromium (IV) content that is mainly toxic to organisms
and can result various types of disease such as in ulceration and perforation of the mucous
membranes of the nasal septum, irritation of the pharynx and larynx, asthmatic bronchitis,
bronchospasms and edema to human being.

RECOMMENDATION
1. Try to detect another heavy metal that may contain in sample water which can cause
harmful to organisms such as copper (II).
2. Tested the same value of concentration of Chromium (VI) more than once in order to
get the average result.
3. To make sure the accurate reading, any finger print, crash or deposited on the surface of
cuvette must be avoided. This is because it can disturb the absorption of light by the
aqueous heavy metal.

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REFERENCES
1. Helmenstine, A. M. Concentration Units & Dilutions (2003).
2. Ingle J. D. J. and S. R. Crouch, Spectrochemical Analysis, Prentice Hall, New Jersey
(1988).

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