SPECTROPHOTOMETRIC DETERMINATION OF THE EQUILIBRIUM CONSTANT OF A REACTION

E.C.D. VALLEJERA

DEPARTMENT OF CHEMICAL ENGINEERING, COLLEGE OF ENGINEERING UNIVERSITY OF THE PHILIPPINES, DILIMAN, QUEZON CITY, PHILIPPINES
DATE PERFORMED: JANUARY 9, 2014 INSTRUCTORS NAME: CHARMAINE M. ALCANTARA

ABSTRACT The instrumental technique of spectrophotometry was executed to determine the equilibrium constant Keq of the formation of the blood-red complex [FeSCN]2+ from Fe3+ and SCN-. Standard soultions were prepared in order to calibrate the analytical wavelength at which the solution absorbs. The spectrophotometer was calibrated with blank solutions to mimic the solution without the analyte present. The absorbances were plotted versus the molar concentrations of [FeSCN]2+ at equilibrium. The equation of the calibration curve was used to acquire the concentration of [FeSCN]2+ of the unknown solutions. The average Keq was determined to be 319.2 which deviates greatly from the theoretical Keq of 890 due to certain errors from the experiment.

INTRODUCTION Spectrophotometry is an instrumental technique used to identify and measure concentrations of specific substances using a spectrophotometer. A spectrophotometer simply measures the intensity of a light beam after it passes through a solution or the quantity of light that is not transmitted or reflected. It consists of two parts, a spectrometer and a photometer. The cuvette which contains the solution of the analyte is placed in such a way that it is found in between the spectrometer and the photometer. The spectrometer produces light of any chosen wavelength which passes through the solution and the photometer measures the intensity of the light. The concentration of a substance which gives color to a solution can be 1

determined by relating its concentration in molarity to the absorbance of light at a specific wavelength which are directly proportional to each other. Beer-Lamberts Law relates the concentration of a colored substance in a solution with its absorbance. (1) Where A corresponds to the measured absorbance of the analyte, is the wavelength-dependent molar absorptivity coefficient, b matches the path length used, and c corresponds to the concentration of the analyte. The absorbance of the analyte can be determined using the transmittance or the ratio of the intensity or power of light that passed through the substance and the power of light before passing through.

With the concentration of the substance known, the equilibrium constant Keq can be determined by obtaining the ratio of the concentration of the products and the reactants of the analyte at equilibrium.
(2)

a 100mL volumetric flask. It was then diluted to mark with the 0.1M HCl solution. 50mL of 0.2M FeCl3 stock solution was prepared by weighing first 1.62g of solid FeCl3 in a 50mL beaker. 20mL of the 0.1M HCl solution was then added and stirred to dissolve the solid FeCl3. The solution was quantitatively transferred into a 50mL volumetric flask and was diluted up to mark with the 0.1M HCl solution. Lastly, 100mL of 0.002M FeCl3 stock solution was prepared by transferring 1.00mL of 0.20M FeCl3 into a 100mL volumetric flask and was diluted up to mark with 0.10M HCl. Preparation of the Standard Solutions Six 6-inch test tubes were prepared and properly labelled as Blank, and Standards 1, 2, 3, 4, 5 to be contained with the solutions mentioned in the table (Table 1) below. Table 1. Components of the Standard Solutions Solution 0.002M FeCl3 Blank 0 mL Standard 0.10 mL 1 Standard 0.25 mL 2 Standard 0.50 mL 3 Standard 1.00mL 4 Standard 2.00 mL 5 0.20M KSCN 1.00 mL 1.00 mL 1.00 mL 1.00 mL 1.00 mL 1.00 mL 0.10M HCl 9.00 mL 8.90 mL 8.75 mL 8.50 mL 8.00 mL 7.00 mL

(3) Where corresponds to the concentration of FeSCN2+, is the 3+ concentration of Fe and is the concentration of SCN all at equilibrium. The experiment aims to calculate the Keq of a reaction, specifically the formation of FeSCN2+, using spectrophotometric techniques. METHODOLOGY Preparation of Stock Solutions Stock solutions were prepared for the components of the standards solutions to be used for the calibration of the spectrophotometer. 1000mL of 0.1M HCl stock solution was first prepared from 12.1M HCl in a 1-L volumetric flask as a solvent for the other preparations for the whole experiment. It was prepared by diluting approximately 8.3mL of 12.1M HCl with distilled water up to mark. 50mL of 0.2M KSCN stock solution was prepared by first weighing 0.97 grams of KSCN in a 50mL beaker. 20mL of the 0.1M HCl solution was then added into the beaker and stirred to dissolve the solid KSCN. The solution was then quantitatively transferred into a 50mL volumetric flask and was diluted to mark with 0.1M HCl. 100mL of 0.002M KSCN stock solution was prepared by transferring 1.00mL of 0.2M KSCN solution that was prepared earlier into 2

The blank, together with standards 1 to 5, were prepared by mixing the aforementioned concentrations of the needed stock solutions in the 6-inch test tubes. The test tubes were then covered with parafilm for the solutions not to evaporate or spill. Preparation of the Unknown Solutions

Four 6-inch test tubes were prepared and properly labelled as Blank, and Unknowns 1-3 to be contained with the solutions mentioned in the table (Table 2) below. Table 2. Components of the Unknown Solutions Solution Blank 0.002M FeCl3 0 mL 0.002M KSCN 5.0 mL 5.0 mL 5.0 mL 5.0 mL 0.1M HCl 5.0 mL 2.0 mL 1.0 mL 0.0 mL

holder, with the clear side facing the spectrometer and the photometer. Then the absorbance was determined and recorded. The cuvette containing the blank solution was then removed from the sample holder and the solution was then placed in a beaker used as a vessel for the discarded wastes. The cuvette was rinsed again three times with distilled water and with the standard solution to be used two times. It was placed into the sample holder and the absorbance was determined and recorded. The same procedure was used for the remaining standards 2 to 5. The absorbance for the unknown solutions were determined by first determining the absorbance of the blank solution. The cuvette used for the standards were first rinsed with distilled water three times and with the blank solution twice. The blank solution was then placed in the cuvette and was placed in the sample holder of the spectrophotometer. The absorbance was determined and recorded. The unknowns 1 to 3 were then tested for their absorbance by first rinsing the cuvette containing the blank solution with distilled water three times and with the unknown solution two times. The unknown was then placed inside the cuvette and into the sample holder of the spectrophotometer. The absorbance was then determined and recorded. RESULTS AND DISCUSSION The stock solutions were prepared by diluting or dissolving them with 0.10M HCl that was prepared from 12.1M HCl. Hydrochloric acid was utilized in the experiment as a solvent for the stock solutions in order to prevent the hydrolysis of the ferric ion (Fe3+). When water is used as the solvent, the ion rapidly hydrolyzes into the [Fe(H2O)5(OH)]2+ as shown in Equation 1. This complex will give the solution a yellow3

Unknown 3.0 mL 1 Unknown 4.0 mL 2 Unknown 5.0 mL 3

The unknown solutions, together with the blank, were prepared by again mixing the aforementioned concentrations of the needed stock solutions in the 6-inch test tubes. They were covered with parafilm after. Calibration of Spectrophotometer the UV-Vis

Before the start of the calibration of the spectrophotometer, the solutions prepared beforehand were all shook in order to mix the contents of the test tubes. The spectrophotometer was calibrated first by finding the analytical wavelength which will be used for the testing of the absorbance of the solutions. Standard 5 was used to find the wavelength where the solution has the greatest absorbance because of the high concentration of FeSCN2+ it contains. The blank solution was first tested for its absorbance by rinsing the cuvette first with distilled water three times and with the blank solution two times from Table 1. The blank solution was then placed in the cuvette. The cuvette was placed inside the sample

brown color which oxidation state of iron.

accompanies

this
(4)

The color of the complex will interfere with the determination of the absorbance of the blood-red FeSCN2+ because the absorbance is directly related to its color, which the color of the yellow-brown [Fe(H2O)5(OH)]2 would greatly interfere with. Adding another color will also absorb light which would correlate to an increase in absorbance and give a false positive.

substance corresponds to the wavelength of light absorbed complementary to it. Referring to figure 1, the solution significantly absorbs the colors opposite the blue area of the color while the blue wavelengths are not absorbed by it. The color shown by the solution is therefore the wavelength of light not absorbed or simply reflected by it. Absorbance should be measured at the analytical wavelength because at that wavelength is where absorbance is at maximum. All wavelengths opposite or far from the analytical wavelength are reflected and using those wavelengths will produce a lower reading for absorption. The blanks used for the calibration of the UV-Vis spectrophotometer were for determining the absorbance of the substance without the analyte FeSCN2+ present in the solution together with the cuvette that can contribute to the absorption of light, thus interfering with the results later on. An excess amount of KSCN was added in order to expend any excess Fe3+ present in the system and assure the correct ratio of the amounts of Fe3+ ions to FeSCN2+ determined from the net ionic equation of the reaction.

Figure 1. Color wheel and the corresponding wavelengths Using the HCl as the solvent for the unknown and standard solutions as well as for the blanks, the spectrophotometer was utilized. The solution with the highest concentration, specifically Standard 5, was used as the basis for determining the maximum wavelength or . This is because concentration is directly proportional to the absorbance of the solution, therefore the solution with the highest concentration of FeSCN2+ ions was used. The analytical wavelength for FeSCN2+ was found to be at maximum 466nm, or around the blue area of the color wheel which is between 430 to 480nm. This certifies the validity of the experimentally-determined analytical wavelength as the color of the 4

The concentration of FeSCN2+ at equilibrium is assumed the same as the initial concentration of Fe3+ because Fe3+ is the limiting reactant in the reaction and any excess ions were expended with the excess KSCN added. These concentrations were used as the independent variables for determining the equation of the best-fit curve. The dependent variable used was the experimentally-determined absorbance of [FeSCN]2+. Initial concentrations of the reactants were first determined before the ICE table was used. The initial concentrations of Fe3+ and were determined and tabulated in Table 4. The solutions for the initial concentrations SCN-

of Fe3+ and SCN- are found in the calculations section of the appendix. Table 3. Initial concentrations of Fe3+ and SCNUnknown 1 2 3 [Fe3+ ]init 6x10-4 M 8x10-4 M 1x10-3 M [SCN-]init 1x10-3 M 1x10-3 M 1x10-3 M

Comparing the experimentallydetermined molar absorptivity constant with a theoretical of 3550 M-1 cm-1, a percent error of 32.90% is found. This is due to the fact that many possible sources of errors are present when the experiment was done. Proceeding from the calibration of the spectrophotometer, the concentration of [FeSCN]2+ at equilibrium was determined by finding their absorbance and using the equation of the best-fit line from the abovementioned calibration to obtain the concentration of [FeSCN]2+ with the absorbance. A new blank was used and the concentrations of KSCN were decreased for this part. The concentrations of KSCN used for the second part of the experiment were decreased from 0.20M to 0.002M. This was done for equilibrium to set in the reaction. Having a large amount of KSCN would result to the right side being favored by the reaction but since the concentrations of KSCN were lessened, equilibrium would be established and Keq can now be determined. Because the concentration was changed for one reactant, a new blank must also be used to compensate for the change. Using the equation of the best-fit line and entering the absorbance from unknowns 1 to 3, the concentrations were determined and tabulated in Table 3. The solutions for the concentrations of [FeSCN]2+eq are found in the calculations section of the appendix. Table 4. Concentration of [FeSCN]2+eq Absorbance Concentration (M) 0.140 1.346x10-4 M 0.179 1.672x10-4M 0.215 1.972x10-4M From the concentration of [FeSCN]2+, the value of the equilibrium constant Keq can be calculated using the ratio of the concentrations of products and reactants at equilibrium. The initial concentrations of the products and reactants can be computed 5

Absorbance vs. [FeSCN]2+eq

Absorbance
0.6 0.4 0.2 0 0 0.0002 0.0004 0.0006 Linear (Series1) Series1

[FeSCN2+]eq

(M)

y = 1197.1x 0.0211 R = 0.9918

Figure 2. Absorbance vs [FeSCN]2+eq As the absorbance versus the concentration of FeSCN2+ was graphed above in Figure 2, the graph shows clearly that the data is linear with its coefficient of linearity being 0.9918. The equation of the best-fit curve, given by linear regression is: (6) Assigning the slope of the best-fit curve as the product of the molar absorptivity constant , the path length b, and the yintercept as the instrument error or the correction factor for the errors found in the experiment, the slope is now used to define the molar absorptivity of [FeSCN]2+. The path length used for the experiment was 0.5cm. Using the path length of 0.5cm and the slope or the product of the molar absorptivity constant and the path length b, the molar absorptivity constant was found to be 2382.2 M-1 cm-1.

using an ICE (initial, change, equilibrium) table. Using the equilibrium concentrations from the ICE table and substituting them into the relationship of the equilibrium constant Keq with the concentrations of products and reactants, Keq can now be determined from equation 2. This system of solving for the Keq was also applied to unknowns 2 and 3. This gives a Keq of 334.2, 317.3, and 306.0. The solutions for the Keq of unknowns 2 and 3 are found in the calculations section of the appendix. The equilibrium constants Keq for unknowns 1 to 3 were determined and tabulated in Table 5. Table 6. Keq of Unknows 1 to 3 Unknown 1 2 3 Keq 334.2 317.3 306.0

Table 7. Sources of Errors Source Type of Error

Fingerpr Determinate, ints on Systematic the cuvette Bubbles Determinate, in the Random cuvette Cuvette Determinate, was not Systematic properly rinsed with the solution to be tested Cuvette Determinate, was not Systematic placed in the right direction Assumpt Determinate, ion of Gross the complete exhausti on of Fe3+ CONCLUSION AND RECOMMENDATIONS The experiment yielded a Keq which deviates greatly from the theoretical value of Keq. This may be caused by a number of errors that may have been committed in performing the experiment. An example of the error is the assumption of the complete reactions of ferric ions in the system at calibration to yield the complex ion [FeSCN]2+. The system may have yielded more or less amounts of the product than what was actually assumed which results txo the great deviation of the experimental Keq. 6

The average of the three equilibrium constants from the unknowns was used as the Keq for the formation of [FeSCN]2+. (7) Solving for the average Keq for the reaction results to 319.2. Comparing the experimental Keq +with the theoretical Keq or the complex-ion formation constant Kf of [FeSCN]2+ at 890, giving a percent error of 64.13%. A percent error of this great magnitude may arise from one or more possible errors from this experiment. One major factor that may contribute greatly to the huge error is the assumption from the Standard solutions that the consumption of Fe3+ is complete.

Another may have been the improper handling of the cuvette which could have led to marks of fingerprints being left on the clear side of the cuvette. This results to a lesser reading of the light that was analyzed by the spectrometer because the mark also absorbs light. Or the presence of bubbles in the solution can also interfere with the result of absorbance of the analyte. The experiment could have given results with less deviation from the actual values if these random errors were to be avoided, especially those that could greatly interfere with the absorbance. The experiment could have also yielded a closer result if proper rinsing of the cuvette was implemented with every determination of the absorbance of a solution. Contamination of solutions could have interfered with the result of the experiment. Overall, the experiment was not successful because of the sheer magnitude of error that the experiment yielded compared to the theoretical value of the equilibrium constant. Factors such as mentioned could have affected the readings in such a way that they resulted to a great deviance to the result. The experiment could have been performed with great care and consideration to many factors that could affect the sensitive and analytic nature of the procedure which would give readings near the exact or correct values. Also, proper handling of the cuvette should be put into consideration if the experiment is to be effective. REFERENCES [1] Petrucci, R.; Herring, F.; Madura, J.; Bissonnette, C. General Chemistry: Principles and Modern Applications; Pearson: Toronto, 2011. [2] Silberberg, M. Principles of General Chemistry; McGraw-Hill: New York, 2010. 7

[3] Inorganic Compounds in Aqueous Solution Colours. http://www.creativechemistry.org.uk/alevel/module5/document s/N-ch5-07.pdf (accessed 20/1/14) [4] Introduction to Coordination Chemistry. http://www.uncp.edu/home/mcclurem/cour ses/chm226/introduction_Coordination_Che mistry.pdf (accessed 20/1/14). [5] Caprette, D. Principles of Spectrophotometry. http://www.ruf.rice.edu/~bioslabs/methods /protein/spectrophotometer.html (accessed 20/1/14). [6] Spectrophotometric Analysis. http://www2.bren.ucsb.edu/~keller/courses /esm223/Spectrometer_analysis.pdf (accessed 20/01/14). [7] Tissue, B. Beer-Lambert Law. http://www.files.chem.vt.edu/chemed/spec/beerslaw.html (accessed 20/01/14). [8] What is the difference between absorbance and transmittance?. http://www.uwplatt.edu/chemep/chem/che mscape/labdocs/catofp/measurea/concentr/ spec20/abstrans.htm (accessed 20/01/14). [9] Chapter 11 Spectrophotometry. ftp://ftp.nmenv.state.nm.us/www/swqb/UO CP/StudyManuals/WWLabStudyGuide/11.pd f (accessed 20/01/14). [10] Spectrophotometry. http://www.usna.edu/ChemDept/_files/docu ments/manual/ApdxI.pdf (accessed 20/01/14). [11] Chemical Equilibrium. http://www.lahc.edu/classes/chemistry/aria s/Lab4Equilibriasp11.pdf (accessed 27/01/14) APPENDIX

I. SAMPLE CALCULATIONS a. Initial concentrations of Standards [FeSCN]2+eq = [Fe3+]init

y = 1191.1x-0.0211. where x corresponds to concentration of complex

Table 1. Components of the Standard Solutions Solution 0.002M FeCl3 Blank 0 mL Standard 0.10 mL 1 Standard 0.25 mL 2 Standard 0.50 mL 3 Standard 1.00mL 4 Standard 2.00 mL 5 0.20M KSCN 1.00 mL 1.00 mL 1.00 mL 1.00 mL 1.00 mL 1.00 mL 0.10M HCl 9.00 mL 8.90 mL 8.75 mL 8.50 mL 8.00 mL 7.00 mL Initial Change Equilibr ium Fe3+ [Fe3+ ]init SCN[SCN-]init [FeSCN]2
+

c. Equilibrium concentrations Table 5. ICE Table

0M +x 0 +x M

-x -x [Fe3+ ]init x [SCN-]init M x M

= 0M = 0.02M =0.00002M =0.00005M =0.0001M =0.0002M =0.0004M b. Concentration absorbance of complex using 8

Unknown 1: Because [FeSCN]2+eq = x, x = 1.346x10-4 M, [Fe3+ ]eq = [Fe3+ ]init x M = 4.654x10-4 M [SCN-]eq = [SCN-]init x M = 8.654x10-4 M

Keq = 334.2 Unknown 2: Because [FeSCN]2+eq = x, x = 1.672x10-4 M [Fe3+ ]eq = [Fe3+ ]init x M = 6.328x10-4 M [SCN-]eq = [SCN-]init x M = 8.328x10-4 M

Keq = 317.3 Unknown 3: Because [FeSCN]2+eq = x, x = 1.972x10-4 M [Fe3+ ]eq = [Fe3+ ]init x M = 8.028x10-4 [SCN-]eq = [SCN-]init x M = 8.028x10-4 M

Keq = 306.0 d. Average Keq

e. Percent error of Keq