ABSTRACT / SUMMARY

Chromium (VI) 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. This health -risky situation has lead to many studies and
investigations on determining the presence and q uantity of chromium in
biological and environmental samples. In the present study, an absorption
method for the determination of chromium (VI) concentration is proposed
and the method is operated spectrophotometrically by tracing the
presence of chromium ( VI) at a wavelength of 435 nanometres. The
concentration of Chromium (VI) in the lake water sample tested is 49.8
parts per million. The experiment is completed and successfully
conducted.

INTRODUCTION

Many heavy metals, such as chromium, are undeniabl y toxic at low

aqueous solution concentrations. Chromium ions are common ly found in
either trivalent, Chromium (III), or hexavalent, Chromium (VI), states.
Chromium (VI) formation is favoured by an oxidizing environment, while
Chromium (III) is favoured 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.

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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 a lots of
studies that have been done profusely in order to determine the health risky chromium in environmental as w ell as biological samples.

AIMS / 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 a nd agriculture purposes.

THEORY

One common source of chromium contamination in natural waters is

automobiles as it is 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 unfavourably permeate
into any water streams nearby via flowing rainwater. Lakes and water
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streams which is 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 mere

concentrations, a lot of federal environmental agencies are doing
researches to determine its presence in many environmental samples.
Thus, in this experiment, absorption spectroscopy w ill be used to detect
low level concentrations of Chromium (VI) in a lake water sample.

As a matter of fact, coloured aqueous solutions have c hemical

species contents which absorb significant wavelengths of light. Similarly,
heavy metals can be identified via absorption of wavelengths of light.
Furthermore, 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 i t 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. Every chemical species absorbs distinct wavelengths of light.
In absorption spectroscopy, the wavelengths 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. Fi ve standard
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solutions need to be prepared from the concentrated Chromium (VI)

standard solution. To calculate the dilute solutions volum e, the solution
dilution formula can be used, as what is written below:
(M 1 ) (V 1 ) = (M 2 ) (V 2 ) ....................(1)

Quantitative analysis using spectrophotometer is based on Beer Lambert Law:

A = LC
where
A

= absorbance value (dimensionless, thus no units is

represented)

= molar absorbance (L / mol.cm)

= path length of the cuvette in which the sample is contained

(cm)

= concentration of the compound in solution (mol / L)

Beer-Lambert Law states that absorbance value depends on the total

quantity of the absorbing compound in the light path through the cuvette.
Thus, if we plot a graph of absorbance versus concentration of the
compound solution, we get a straight line passing through the origin
(0, 0). 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 compoun d with a high molar absorbance can be easily
detected.
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APPARATUS / REAGENTS

Distilled water

spectrophotometer

10 millilitres square cuvette bottle

pipette

5 set of 10 millilitres of 1, 20, 60, 85 and 100 p.p.m of diluted

solution of Chromium (VI)

5 set of 300 p.p.m of standard Chromium (VI) solution of 0.03, 0.67,

2.00, 2.83 and 3.33 millilitres.

lake water sample.

EXPERIMENTAL PROCEDURE

1. All the materials and apparatus are set up.

2. The 10 millilitres of square cuvette bottl e is filled up with distilled water
and placed on the spectrophotometer slot. A zeroing step is done to
ensure that all the data obtained later is accurate.
3. 5 series of diluted solution are prepared by using the diluted method
with distilled water with the range concentration of 1 until 100 parts per
million (p.p.m) solutions. (1 p.p.m, 20 p.p.m, 60 p.p.m, 85 p.p.m and 100
p.p.m)

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4. 10 millilitres of each series of diluted solution is poured into square

cuvette bottle and placed on the spectrophotometer slot to check the
absorbance value by using the spectrophotometer.
5. All the data obtained is recorded.
6. 10 millilitres of lake water sample is poured into the square cuvette
bottle and spectrophotometer is used to determine the Chromium (VI)
concentrat ion contained in the water sample collected.
7. All the value is analyzed; the observation and the data obtained are
recorded.

RESULTS
Volume of Chromium
(VI) , millilitres

0.03

0.67

2.00

2.83

3.33

20

60

85

100

0.008

0.131

0.342

0.498

0.594

Concentration of
Chromium (VI), parts
per million
Absorbance Value

Table 1.1 Determination of absorbance value using spectrophotometer

Lake W ater Sample

Absorbance Value

Trial 1

0.247

Trial 2

0.257

Trial 3

0.256

Average value :

0.253

Table 1.2 Determination of absorbance value of water sample using

spectrophotometer

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Graph of Absorbance Value versus Concentration of Cr (VI) in ppm

Absorbance Value

0.65
0.6
0.55
0.5
0.45
0.4
0.35
0.3
0.25
0.2
0.15
0.1
0.05
0

0.594

R = 0.9987
0.498

0.342

0.131

0.008
0

20

40

60

80

100

Concentration of
Chromium (VI) in
120 unit of p.p.m

Graph 1.1 Graph of Absorbance Value versus Concentration of

Chromium (VI) in units of p.p.m

SAMPLE CALCULATIONS

To dilute 10 millilitres standard solution of 1 p.p.m, the solution dilution

formula is used in order to calculate the volume of chromium (VI) solution.
(M 1 ) (V 1 ) = (M 2 ) (V 2 ).................... (1)
(300 ppm) (V1) = (1 ppm) (10 mL)
V 1 = 0.03 mL
All the volumes of chromium (VI) solution nee ded to dilute each standard
solution are stated in Table 1.1.

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By interpolating Graph 1.1, the equation of the linear line is given as;
y = 0.005 x + 0.004

By substituting y with the average of absorbance value of water sample ,

which is 0.253, hence we get that;

= y 0.004
0.005
= 0.253 0.004
0.005
= 49.8

Notice that x is equivalent to the concentration of Chromium (VI) contains

in the water sample, which is 49.8 parts per million.

DISCUSSION

This experiment is conducted to achieve a few objectives, which 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 drink ing water and agriculture
purposes.

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The lake water sample used to test for presence of chromium is

believed to be contaminated as the lake is situated near a roadway where
road users with automobiles use it every day. As mentioned before, 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.

According to the values of concentration of chromium (VI ) which is

apparently calculated to contain 49.8 parts per million , this obviously
means 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 lake water sample
tested is then concluded to be unsafe and unhygienic for drinking water
for public consumption 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. T his might lead to difficulties when
measuring the volume of chromium (VI) needed to dilute the standard
solutions.

Besides that, 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 oute r surface of the cuvette can become an
unfavourably interference for the solutions to absorb wavelengths of light

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in the spectrophotometer. Thus, the reading of absorbance by the

instruments may not be accurate.

In addition, the experiment conducted to d etermine 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 mig ht 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.

CONCLUSION

The concentration of Chromium (VI) in the lake wa ter sample tested

is 49.8 parts per million. Since it is more than 0.100 parts per million as
what has been standardized by state and federal regulatory agencies, the
water sample is concluded to be contaminated and thus, not suitable for
drinking or agricultural purposes.

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RECOMMENDATIONS

In order to get more accurate values, a few recommendations and

precautions must be considered when performing the experiment.

Firstly, in this experiment, selection of wavelength value of light in

the spectrophotometer is quite essential. To get an ideal value, it is
preferably to use a wavelength where the component absorbs
substantially.

Next, avoid leaving a fingerprints or any kind of smudge on the outer

surface which will corrupt the reading of a bsorbance value of
spectrophotometer.

Besides that, the determination of the absorbance value of each

diluted solutions must be done at least 3 trials. By doing so, more
accurate values can be obtained.

Likewise, the square cuvette bot tle must be clea nsed thoroughly and
properly.

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REFERENCES

1. Ressalan S, Chauhan RS, Goswami AK, Purohit DN. (1997) Review of

spectrophotometric methods for determination of chromium. Rev. Anal.
Chem. 16 (2), 69-171.
2. 13. Mohamed AA, Ahmed SA, El -Shahat MF. (2001) C atalytic spectrop hotometric determination of hexavalent chromium. J. Trace Microprobe
Tech. 19 (2), 297 -311.
3. www.googlebooks.com/experimentingeneralchemistryf rommeasurenet

RECOMMENDATIONS

Refer to the attachment provided on the next page.

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