Linda Christian

58125 CHEM
1411- 009

Spectrophotometric Analysis
of a Cobalt (II) Chloride Solution

General Chemistry I
(Chemistry 1411)

Professor ………….

By

………………

November 28, 2011

Linda Christian
58125 CHEM
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Spectrophotometric Analysis of a Cobalt Ion Solution

ABSTRACT:

State the purpose: The aim of this experiment was to determine the absorption spectrum of Cobalt (II)

ion using a spectrometer and Beer’s Law Plot. State the compound or substance studied: This

experiment was performed using pre-diluted Cobalt (II) Chloride solution of known concentration and

pre-diluted Cobalt (II) chloride solution of unknown concentration. State the method or specific sections

that were studied in the experiment. The absorption spectrum of the cobalt (II) ion was determined by

calibrating the spectrometer with a cuvette that contained distilled water, then measured the

absorption at different wavelength in the visible light region using a cuvette which contained 0.150M

Cobalt (II) Chloride, CoCl2. State the results: The concentration of the Cobalt chloride was determined to

be 0.09322M at 500nm on a graph using the Beer’s Law Plot and by the method of least squares to get

the slope which is k = 2.93.

INTRODUCTION:

Background on the Spectrophotometer and theory of light absorption: In order to investigate

either the absorbance or transmittance of light with a series of solutions given of Cobalt (II) ‘t

use personal pronounsChloride (CoCl2) and to determine the unknown solution you will need a Comment [AL1]: No personal pronouns
throughout the text

device called spectrophotometer. This instrument detects the amount of radiant light energy that

is absorbed by the molecules. In order for the spectrophotometer to do this it must have five

basic components: a light source, a diffraction grating, a slit, a photoelectric tube, and a digital

meter. There are two adjustments before the light-absorbing properties of a solution can be
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measured must be done to the spectrophotometer. First, the diffraction grating must be adjusted

for desired wavelength of light passes through slit. This is like adjusting the amount of light you

want to come in a room with the venetian blinds. Secondly, the output of the phototube must be

calibrated. Here is why it is important to clean your test tubes with lint free wipes to remove any

finger prints or water droplets on test tubes. Now you are ready to test your samples given and

determine the unknown. Remember to record your results onto a table then graph, zero out

before each testing, and do not contaminate your test tube with impurities either in the solution

or on the outside glass surface during the experiment.

The Beer-Lambert Law: The theory behind this method of analysis is called the Beer-Lambert

Law (1). Since the light that enters the cuvette has a certain strength or Intensity, it will come out

with a different Intensity depending on the substance inside the cuvette. If the substance is more

concentrated, it absorbs more of the light and allows less to be transmitted. There is a correlation

between the amount of light absorbed and the concentration of the solution, all other factors

being kept constant as possible.

Formally speaking, the absorbance of light by a solution is proportional to the concentration of

the compound in the solution and the thickness of solution that the light must pass through. The

relationship is expressed in the general equation of the Beer-Lambert law:

A = abc (1)

where A = absorbance, a = a constant for the substance being studied (also known as €),

b = the thickness of solution the light passes through, and c = the concentration of the substance.
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Since the same instrument will be used throughout the analysis on the same compound, a and b

are combined together into one constant: k = ab. The equation is rewritten as:

A = kc (2)

Before the analysis, k must be determined for the instrument and the compound that is being

analyzed. Absorbance readings (A) are taken for a set of solutions of known concentration (c)

and plotted with Absorbance on the y-axis and concentration on the x-axis. This way, k can be

calculated from the slope of the graph:

k = A/c (3)

Mathematical methods can be used here. It is also important to make sure that the instrument is

set to the wavelength at which the compound absorbs highest and therefore gives best results for

the readings. This is called the maximum wavelength or lambda max, where lambda is the

symbol for wavelength (λmax).

For the same reason, the dilution of the unknown solution is selected to be the one that gives the

highest absorbance reading at lambda max.

Alternative methods or calculations if any: Percent transmittance, %T, is sometimes measured

instead of absorbance. On instruments with scales rather than digital displays there is less error

associated with reading the transmittance. It is also more directly related to the light level being

measured. The absorbance and the percent transmittance are related as follows:

A = -log (%T/100) (4)

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Once all of these conditions are met, a reliable analysis can be made. Spectrophotometry is used

in many biology and biotechnology applications: for food dyes, protein in milk and protein

drinks, rates of photosynthesis, sugar enzyme reactions, and track the growth of bacterial culture

(2).

The Purpose of the Experiment in connection to the Theory: Using a set of solutions of Cobalt

(II) Chloride (CoCl2) we will investigate the absorption of light. We will be given a solution of

this substance whose concentration is unknown but too large for accurate measurement of the

absorption of light. The task is to determine the concentration of this solution. Recall that Beer’s

law states that the absorbance is directly related to the concentration (c) of the substance that

absorbs light, or where k is a constant:

A = kc (2)

The equation for Beer’s law has the same form as the equation for a straight line:

Y = mx + b (2a)

A comparison of these two equations, 2 and 2a, above indicates that:

y = A, x = c

m = slope = k

b = intercept on y axis = 0

A straight line should be obtained when we plot the absorbance at various concentrations against

the concentrations. Calculations of concentration will be made from those readings and discussed

in detail in the Data and Calculations sections.

Linda Christian
58125 CHEM
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PROCEDURE

Explain steps in Preparation: The spectrophotometer was turned on to warm up before the

solutions were prepared. 50mL of a stock solution of 0.150M cobalt chloride was added into a

100mL beaker. After that, 50mL of deionized water was placed into another 100mL beaker. By

using a 10.00mL pipet, 5.00mL of 0.150M Cobalt Chloride solution was transferred into test

tube 5 from the 100mL beaker;

For the rest of the solutions, 4.00mL of 0.150M Cobalt Chloride solution was transferred into

test tube 4 from the 100mL beaker; 3.00mL of this solution was transferred into test tube 3;

2.00mL of this solution was transferred into test tube 2 and 1.00mL of this solution was

transferred into test tube 1. Also, 5.00mL of deionized water was added into test tube 0 from the

100mL beaker by using another 10.00mL buret. After that, by delivering with the buret, 1.00mL

of deionized water into test tube 4, 2.00mL of deionized water into test tube 3, 3.00mL of

deionized water into test tube 2, and 4.00mL of deionized water into test tube 1. After the

deionized water was added to the 0.150M Cobalt Chloride solution of test tube 1-5, these test

tubes were mixed thoroughly.

Explain steps in Instrumental Analysis: Preparing the spectrophotometer: Through following the

number order, those mixed solutions of test tube 1-5 were transferred into cuvette 1-5 by using a

10.00mL pipet. The deionized water of test tube 0 was also transferred into cuvette 0 by using a

10.00mL pipet. After all the preparations of solutions were done, the displayed wavelength on

the spectrophotometer was set to 430nm and also the cuvette 0 was inserted into the

spectrophotometer. Then the blank button was push for setting the 0.00 absorption. The

absorption of cuvette 5 in 430 nm was recorded by replacing the cuvette 0 to cuvette 5 inside of
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the spectrophotometer. After that, the displayed wavelength on the spectrophotometer was

changed to 460nm and also the cuvette 0 was inserted into the spectrophotometer again for

setting the 0.00 absorption.

Determining the Wavelength of maximum absorption:The absorption of cuvette 5 in 460 nm was

recorded by replacing the cuvette 0 to cuvette 5 inside of the spectrophotometer. By changing the

displayed wavelength on the spectrophotometer and repeating above steps, the absorptions of

cuvette 5 in 480 nm, 500 nm, 510 nm, 520 nm, 540 nm and 570 nm were recorded one by one.

Through compare the absorption of each wavelength, the wavelength of highest absorption was

selected for the second part of the experiment---unknown identification.

Explain steps in unknown analysis: The displayed wavelength on the spectrophotometer was set

to the selected one which was recorded from previous experiment. And the cuvette 0 was

inserted into the spectrophotometer for setting the 0.00 absorption. After this, a Cobalt Chloride

solution with unknown concentration was obtained from instructor and the cuvette 5 was washed

by deionized water. Then the Cobalt chloride solution with unknown concentration was

transferred into this cuvette 5. The absorption of this solution was recorded by replacing the

cuvette 0 to the cuvette 5 inside of the spectrophotometer. Then the concentrations of cuvette 1-4

were calculated and recorded. Finally the absorptions of cuvette 1-4 were recorded by replacing

the cuvette 5 to all of them.

EXPERIMENTAL DATA AND CALCULATIONS:

A. Data:
Part 1. Determining Wavelength of Maximum Absorption
Table 1. Absorption spectrum of standard Cobalt ion solution
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Wavelength (nm) absorption Wavelength (nm) absorption

430 0.1662 520 0.7889

460 0.4939 540 0.5148

480 0.6328 570 0.1396

500 0.8180

Fig 1. Graph of A (y axis) vs. concentration and determine the optimum wavelength.
Linda Christian
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Based on this data in the graph , the highest absorption level is 0.8180 when the wavelength is

equal to 500nm. Therefore, 500nm would be determined as the optimum wavelength for next

step of experiment.

Part 2. Beer’s Law Plot for the Standard Solutions of CoCl2

Solutions were prepared by diluting a known volume from the Stock solution that was 0.15M in

concentration. Absorbance readings were made at 500nm which was the maximum wavelength,

called lambda max (λmax).

Table 2. Concentration and Absorbance Readings for Standard Solutions of CoCl2

Tube # Concentration Absorbance

(moles/L) (A.U.)
1 0.0300 0.1555

2 0.0600 0.3208

3 0.0900 0.4882

4 0.1200 0.6450

5 0.1500 0.8181

Figure 2. Graph A (y axis) vs. c (x axis) and determine the product €b from the slope of the

line. K = €b= 2.93

Linda Christian
58125 CHEM
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Part 3. Identifying the unknown concentration

Unknown identification number: B

Table 3. Trial solutions were prepared according to the dilution on the table and gave the
following Absorbance readings

mL of unknown mL of H2O A

3.5 1.5 0.192

3.0 2.0 0.156

2.5 2.5 0.138

2.0 3.0 0.113

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Table 4. Concentrations were calculated using k for each of the absorbances:

A c diluted unknown, M c undiluted unknown, M

0.192 0.06552 0.09358

0.156 0.05324 0.08873

0.138 0.04709 0.09418

0.113 0.03856 0.09640

Average: 0.093223 M

B. Calculations:

1. Concentrations of the Diluted Standard Solutions

The main formula for the calculation of concentrations was based on:

Mi Vi = Mf Vf (3)

Sample calculations are:

0.150M ×1mL = Mf ×5mL Mf1 = 0.300M

0.150M ×2mL = Mf ×5mL Mf2= 0.600M

0.150M ×3mL = Mf ×5mL Mf3 = 0.900M

0.150M ×4mL = Mf ×5mL Mf4 = 0.120M

2. These concentrations were used to graph Absorbance vs Concentration (Figure 2) and in

calculating the slope by the least squares formula in the next section.
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3. Manual Calculation of Slope from Linear Regression

Calculations were made from the formula for the slope and equal to k , the constant:

k = ∑ cA /∑ c2 = M-1 (5)

where the slope m from equation (2a) is the constant k which is specific for the type of analysis
and the characteristics of the spectrophotometer.

Also, ∑cA is the sum of the products of each Absorbance with its corresponding concentration

and ∑ c2 is the sum of the squares of the concentration.

k = (0.0300) (0.1555) + (0.0600) (0.3208) + (0.0900) (0.4882) + (0.1200) (0.6450) + (0.1500)(0.8181)

(0.0300)2+ (0.0600)2 + (0.0900)2 + (0.1200)2 + (0.1500)2

k = 2.93 M-1

This value of k will be used to find what the concentrations of the unknown diluted solutions are

4. For the concentrations of the diluted unknown solutions using k from linear regression:

c=A/k (6)

Thus:

c1= 0.192 / 2.93 = 0.06857 M c3 = 0.138/ 2.93 = 0.04710 M

c2 = 0.156 / 2.93 = 0.05324 M c4 = 0.113/ 2.93 = 0.03857 M

5. Concentration of Cobalt (II) chloride calculated

The same formula was used for the calculation of concentration of the undiluted unknown –

working backwards to find the concentration of the undiluted original solution, C1:

C1 V1 = C2 V2 (7)
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Calculations were done for the each of the diluted unknown and the final answer averaged.

C1 = C2 V2 / V1 (7) rearranged

C1 = 0.06552 M x 5.0mL / 3.5mL = 0.09358M

C1 = 0.05324M x 5.0mL / 3.0 mL = 0.08873M

C1 = 0.04709M x 5.0 mL / 2.5 mL = 0.09418M

C1 = 0.03856M x 5.0 mL / 2.0 mL = 0.09640M

Average: 0.37289 / 4 = 0.093223 M

Reported average concentration of unknown solution = 0.09322 M

6. Concentration of Cobalt (II) chloride unknown directly from graph on Excel

For comparison:

One method described in the lab manual for determining concentration from absorbance is to

read off the value directly from the graph and use this to figure out the original concentration.

Selected Unknown absorbance = 0.192

Matching Concentration of unknown solution on the graph = 0.0412M

To calculate the concentration of the undiluted unknown with this concentration:

C1V1 / V2 = 0.0412M x 5.0mL / 3.5 mL = 0.0588M which is on the low side and shows that a

more accurate method would be by linear regression. This makes it clear that our experimental

values were not on a perfect straight line.

Linda Christian
58125 CHEM
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DISCUSSION

Summarize what you set to do in the experiment: In this experiment, two things had to be

measured. First, the relationship between absorption and wavelength of radiation that found the

best absorbed wavelength for Cobalt (II) Chloride solution. Second measurement is the

relationship between absorption and concentration of solution which proved Beer’s law.

Discuss determining lambda max: The best-absorbed wavelength was the first factor that

should be figured out in the first part of experiment. Based on data, the 500nm had highest

absorption rate by Cobalt (II) chloride. Below 500nm, the absorption kept increasing and it

decreased when wavelength reached beyond 500nm. Therefore 500nm was the selected

maximum absorbance wavelength for this experiment. It was important because it had to be used

in the second part of this experiment.

Discuss the standard curve, your results and how they will be used: The second part of this

experiment was looking for the relationship between absorption and concentration of cobalt

chloride solution. In fact, the absorption would be increased when the concentration rose.

According data read by spectrophotometer, a function had been calculated to give the equation of

a straight line and yielded a slope = 2.93 Using this for the relationship between absorbance and

concentration: y = 2.93 x. In this function, X represents the concentration of solution and Y is the

absorption of the solution. It proved the Beer’s Law and showed they are directly related

between absorption and concentration.

Discussion of errors and limitation of the experiment: There were two main errors happened

in the experiment which may impact or affect this experiment. They were the actual

concentration of Cobalt (II) ion solution and the spectrophotometer itself.

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First, the actual concentration of Cobalt ion solution may be not same as concentration

showed on label. The laboratory-made solution should very accurate and pure. However, during

reserving and moving those solutions into lab, there have many factors which affect the

concentration of solution by environment. Therefore, the error of the solution used may transfer

into the experiment and caused error. Fortunately, this error should be really small and will not

affect experiment seriously.

Second, the spectrophotometer may have some instrumental errors in this experiment. By

calibrating the instrument and using a blank, we prevented these errors from becoming too big.

This experiment required high accuracy of operation and measurement. Therefore, several

procedures were used in this experiment to avoid error.

What was done further to minimize error: First, throughout the experiment the pipets were

used to measure the volume of solution and water. Because the accuracy is so important, the

experiment should use equipment as accurately as possible. Also, when measuring liquid by

pipet, decimal places had to be read according to the marks on the instrument and may have been

missed.

Second, the cuvettes had to be rinsed with deionized water, and the Cobalt Chloride solution

before they was filled and placed in the spectrophotometer. In addition, the outside of tube was

wiped clean to erase any fingerprint oils on the surface of the cuvettes. Finally, the test tubes

used to dilute the solutions was dried and not rinsed before being used in order to prevent water

droplets from producing an inaccurate result.

Linda Christian
58125 CHEM
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Discussion of determining of the unknown: The same formula was used for the

calculation of concentration of the undiluted unknown – working backwards to find the

concentration of the undiluted original solution, C1:

C1 V1 = C2 V2 (7)

Calculations were done for the each of the diluted unknown and the final answer averaged. The

average concentration calculated for the unknown was 0.09322M. Standard Deviation analysis

should have been done on the results of the analysis.

CONCLUSION

Statement of findings: This experiment showed that Cobalt (II) Chloride solutions in dilute

concentration follow the Beer-Lambert law at 500nm. The Cobalt (II) Chloride standard solution

had highest absorption when the wavelength was 500nm. The method of least squares was

selected over direct reading from the graph. The average concentration calculated for the

unknown was 0.09322M. Assess the method itself: The results showed good correlation. It also

showed that the concentration of an unknown cobalt chloride solution can be determined to a

useful degree of accuracy.

REFERENCES: (add lecture and lab book)

(1) Lab Procedure from Ebbing and Gammon given to class

(2) Internet: http://www.hhmi.org/coolscience/resources/SPT--FullRecord.php?ResourceId=34

(3) Houghton Miffin Company, The absorption Spectrum of Cobalt (ll) Chloride, (Experimental

requirements were obtain from this source)