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 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 for each

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

Concentration (ppm) 20 40 60 80 100

Volume of Cromium (IV) (mL) 6.4 12.8 19.23 25.64 32.05

Absorbance 0.237 0.504 0.751 1.017 1.252

Absorbance value for water sample: 0.324

1.4 1.2 1 Absorbance 0.8 0.6 0.4 0.2 0 0 -0.2 20 40 60 Concentration (ppm) 80 100 120 y = 0.0126x - 0.0051 R = 0.9997

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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