Spectrophotometric Determination of the Equilibrium Constant of a Reaction

J.R.A. Ibale Institute of Chemistry, College of Science University of the Philippines, Quezon City, Philippines Date Performed: January 16, 2013 Instructors Name: Maro R. Pea

ABSTRACT The objective of this experiment, Spectrophotometric Determination of the Equilibrium Constant of a Reaction, is to determine the equilibrium constant, Keq, of the formation of thiocyanatoferrate(III) complex, [Fe(SCN)2+], through spectrophotometry and the application of the Beer-Lambert-Bougert law. Acidified (with HCl) reaction systems of FeCl3 and KSCN were analyzed and the absorbance of the formed [Fe(SCN)2+] was measured using UV-Vis SequoiaTurner Model 340 spectrophotometer. After obtaining the equilibrium values of the concentrations of [Fe3+], [SCN-], and [Fe(SCN)2+], Keq was calculated as well as the percent deviation of the experimental value from the literature value. It was observed that the experimentally determined Keq (331.10) was only 66.67% of the known Keq (890) of the reaction. From the error, it can be concluded that the experiment was a failure which has arisen from experimental inaccuracies. It was then recommended that this experiment is to be done in a fairly consistent ambient temperature using more diluted test reaction systems obtained from reagents with greater purity and that the spectrophotometer must be handled with greater accuracy to yield a more accurate result.

INTRODUCTION Spectrophotometry, a branch of spectrometry, is a method of determining the concentration of substances through quantifying the electromagnetic spectra that atoms of the unknown can absorb. Specifically, spectrophotometry focuses on light intensities that are within the spectrum of visible light, near-ultraviolet, and near-infrared, which is absorbed or transmitted by the sample. [1] This quantitative study particularly uses the device called spectrophotometer. It has two main parts, the spectrometer which emits a specific light wavelength, and the photometer which measures the light intensity that the sample has transmitted. [1] This experiment aims to study the equilibrium constant of a reaction through

analysis by spectrophotometry. It intends to acquaint the students with the spectrophotometer, in particular, as well as with a new method of determining the equilibrium constant of a reaction. Here, spectrophotometry is used to determine the concentration of the reagents in the sample. [2] This value is then used to calculate the equilibrium constant of the reaction. The reaction between iron and thiocyanite is observed for this activity. It has the following system,

In this reaction system, a blood-red complex forms, . This complexs absorption range will be measured by the a UV-Vis spectrophotometer and the concentration can then be obtained through algebraic

manipulation of Beer-Lamberts law (which relates the light absorption of the colored substance in the system to its concentration), A = bc Where A = absorbance, = molar absorptivity coefficient in M-1cm-1, b = path length in cm, and c = analyte molar concentration. Using the calculated concentration the equilibrium concentration can then be computed with the use of the expression, . Although the method in consideration is practical, in general, this is a very limited scheme given that it can only be executed accurately if the solutions are highly diluted and that the reagents used have a high purity. Otherwise, the data will not follow a regressing pattern (linear). Moreover, the over-all experiment must be done in a fairly level atmosphere (i.e. temperature conditions must be precisely the same for all tests), since Keq changes significantly through variable temperature. METHODOLOGY In the experiment, 0.20 M FeCl3, 0.20 M KSCN, 0.002 M FeCl3, and 0.002 M KSCN were prepared. Anhydrous salts were used for the preparation of 0.002 M FeCl3 and KSCN. Five Standard solutions and 3 Unknown solutions, each with a blank solution, were prepared. The solutions were added with 0.1 M HCl as a solvent. The volumes added to the solutions are shown in Tables A and B. Table A. Volume Standard Solutions Solution Blank S1 S2 S3 S4 0.20 M KSCN, mL 1.0 1.0 1.0 1.0 1.0 of Solutions for

S5

1.0

2.0

7.0

Table B. Volume Unknown Solutions Solution Blank U1 U2 U3 0.002 M KCSN, mL 5.0 5.0 5.0 5.0

of

Solutions

for

0.002 M FeCl3, mL 0 3.0 4.0 5.0

0.10 M HCl, mL 5.0 2.0 1.0 0.0

The solutions were covered and left inside the locker for 2 days. Before the absorbance reading procedure was done, the spectrophotometer was allowed to warm up and the analytical wavelength to be used was set to 466 nm. After the orientation as to the proper usage of the apparatus, the spectrophotometer was then calibrated with the standard blank and the calibration curve for the experiment was obtained using the standard solutions. In using the spectrophotometer (with a path difference of 1 cm), the standard solutions were placed in a cuvette (which was rinsed with the analyte to be tested before usage). The cuvette was then placed and analyzed in a UV-Vis Sequoia-Turner Model 340 Spectrophotometer. In order to obtain a corrected absorbance value of the standard solutions, the standards blank solution was placed first and Autozero was performed. Similar procedures were done for the unknown solutions. After the analysis, absorbance readings that were obtained from the standard solutions, a calibration curve was formulated through linear regression analysis. Using the BeerLambert-Bougert law in the form of the experimental calibration curve, the equilibrium values of the complex, FeSCN2+, was obtained. After further calculations for the equilibrium concentrations of the reactants, Fe3+ and SCN-, the equilibrium constant for the reaction was obtained using the following equation,

0.002 M FeCl3, mL 0 0.1 0.25 0.5 1.0

0.10 M HCl, mL 9.0 8.9 8.75 8.5 8.0

. The percent error was then compute with basis on the literature value of the equilibrium constant of this reaction, Keq = 890. RESULTS AND DISCUSSION In the experiment, the reaction system of the formation of monothiocyanatoferrate (III) complex, Fe(SCN)2+ was formulated in terms of standard solutions (solutions with known concentrations) and unknown solutions. The standard solutions were used to calibrate the spectrophotometer as well as to obtain the calibration curve of the set-up used. For the solutions, each stock solution to be used in the experiment proper was acidified with 0.1 HCl. This was done especially for the hydrated ferric ion, FeCl36H2O, since it easily hydrolyses in water. [3] This is due to a property of FeCl36H2O, the hydrated ferric ion. In water, it acts as a Brnsted acid and has the hydrolysis equation, [4] Fe(OH)63+ + H2O [Fe(H2O)5OH]2+ + H3O+ KA = 0.006 (where [Fe(H2O)5OH]2+ can also be Fe(OH)3) [Fe(H2O)5OH]2+ or Fe(OH)3 is a very insoluble compound. Although the high solubility of rust is well known, the acidity of the hydrated ferric ion is not well known which confuses the actual concentration of Fe3+ in the solution and is would become considerably less than the concentration from the start. To control this, the ionization of Fe3+ can be lessened by increasing the acidity of the solution, as in the experiment where 0.1M HCl was added so Fe(OH)3/ FeCl36H2O = 0.006/0.1 = 0.06, i.e., 6% of the total iron is present as the dissociated species (Fe(OH)3) while ~94% is present as Fe3+ (FeCl36H2O). Moreover, if the said hydrolysis cant be contained or inhibited, the absorbance reading would not be isolated on FeSCN2+ alone, but would

have a considerable contribution from the color of rust and or the brown metal-lignin complex, Fe(H2O)5OH2+. After the reagents were prepared, which included 100 mL of 1.0 M HCl solution, 250 mL of 0.1 M HCl (from the 1.0 M stock HCl), 50 mL 0.1 M stock KSCN solution, and acidified (0.1 M HCl) solutions of 0.002 M KSCN (100 mL), 0.2 M FeCl3, and 0.002 M FeCl3, the spectrophotometer was then prepared. The spectrophotometer used in the experiment was the Sequoia-Turner Model 340 Spectrophotometer which had already been turned on to warm up while the solutions were prepared. The wavelength () of the spectrophotometer was turned to 466 nm initially. The same was also used as the analytical wavelength for the entire experiment. The was chosen with basis on the absorptive properties of atoms. It is well known that whatever color a material has would be the same spectra of light that it reflects from its surface. Conversely, all other colors other than the reflected ones are absorbed, primarily its complementary color. Since, Fe(SCN)2+ is a complex whose color is near the spectra of red, then it would be absorbing light with other than those near the red spectrum, especially those near the blue spectrum. (Blue is the complement of red.) The blue spectrum has a that ranges near 450 nm, which is why the analytical used for this study is 466 nm. [5] Before any of the solutions were tested, both standard and unknown, blanks were first used to level the spectrophotometer to zero. This is to account for the light absorbed by the cuvette and the excess SCNwhich lingers in the solution. During the calibration of the spectrophotometer, the standard solutions were first measured for their absorbance in order to set a calibration curve which will also serve as a basis for later calculations in the equilibrium constant determination. The standard solutions contained varying amounts of FeCl3, SCN-, and HCl, wherein

HCL served as a means of acidifying the reactions system, and FeCl3 is the limiting reagent. Note that all of the solutions have the same total volume of 10 mL each. Equivalently, SCN- was added in excess. This is to ensure an ideal condition wherein all of the Fe3+ reacts to form Fe(SCN)2+ and even further as Fe(SCN)63-,[6] making it possible to solve for the best fit line when the data is regressed in a linear fashion (calibration curve of absorbance, Absorbance vs Concentration), further meaning to solve the proportionality constant of the Beer-Lambert-Bougert law, k (also, *b or molar absorptivity*path length). (A = k[Fe(SCN)2+])[2, 3] But even if the system can form other complexes, the experiment only aims to study the absorbance of the first formation constant which is why there was only a controlled amount of excess SCN- that was added. Ideally with this, the reaction system would only yield a single complex which is Fe(SCN)2+. Through calculations, the initial concentrations of [Fe3+]init and [SCN-]ini were obtained. The equilibrium concentration of [Fe(SCN)2+]eq then is just the same as the obtained concentration of [Fe3+]init since it was the limiting reagent in the reaction. Listed in Table 1 are the acquired values. Table 1. Calibration Curve. Soln Abs [Fe3+]init [SCN-]init Std 1 Std 2 Std 3 Std 4 Std 5 0.013 0.057 0.122 0.275 0.482 2.0x10-5 5.0x10-5 1.0x10-4 2.0x10-4 4.0x10-4 0.02 0.02 0.02 0.02 0.02
[Fe(SCN)2+]
eq

obtained. This curve is basically the equation of its best fit line (through linear regression) of the data and is shown in Figure 1.
0.6 0.5 0.4 0.3 0.2 0.1 0 0 0.0002 0.0004 0.0006

Figure 1. Absorbance vs. [Fe(SCN)2+] graph. This graph has the equation y = 1240.1x0.0012. This is similar to Beer-LambertBougert law, A = bc, wherein, given that b (path length) = 1 cm, = 1240.1 M-1 cm-1 and -0.0012 is the experimental aberration which is practically negligible as well. Since the R2 value (0.9926) of the curve does not deviate too far from being a line, the calibration curve is valid. After calibrating the spectrophotometer, the unknown solutions were then tested. The blank solution for the unknown was first tested to recalibrate the base zero of the apparatus since the amount and the concentration of SCN- used in this part differs from that of the standard solutions even if the same cuvette were used. In this part of the experiment, a more diluted concentration of SCN- and Fe3+ is used. Highly diluted regents are used in this part of the experiment in order to isolate the absorbance reading to that of the metalligand complex, Fe(SCN)2+ alone. [7] This is possible because in diluted form, SCN- as well as Fe3+ have negligible, if not at all, absorbance. Moreover, as stated earlier, this type of spectrometry can only yield results that will follow the calibration curve if the solutions to be analyzed are highly diluted and have come from reagents with high purity. Through a similar method of calculation, as with the standard solutions, the initial concentrations of Fe3+ and SCN- were calculated. The values are tabulated in

2.0x10-5 5.0x10-5 1.0x10-4 2.0x10-4 4.0x10-4

After measuring the absorbance of each standard solution, the data was meant to be further mathematically corrected which is done by subtracting the absorbance reading from the blanks, but since the zero for the standards were based on the blank the absorbance readings of the standards can be used immediately, which is also found in Table 1. This was also plotted in a graph from which the calibration curve was

Table 2, together with the actual absorbance readings for unknown solutions. Table 2. Absorbance of Unknown Solutions. Soln Abs [Fe3+]init [SCN-]init Unk 1 0.170 6.0 x 10-4 1.0 x 10-3 -4 Unk 2 0.219 8.0 x 10 1.0 x 10-3 Unk 3 0.257 1.0 x 10-3 1.0 x 10-3 Afterwards, the equilibrium concentrations of Fe(SCN)2+ were then obtained by using the calibration curve which was determined earlier in the study. On the other hand, the equilibrium concentrations of Fe3+ and SCNwere calculated using the ICE table. Since the mole ratio of the reactants and product is 1:1:1, the equilibrium concentrations of Fe3+ and SCN- can be obtained by using the said approach (Table 3). Table 3. ICE Table of Concentrations
[Fe3+] [SCN-]
[Fe(SCN)2+]

When compared to the literature value of Keq = 890, for this reaction system. [3] The %error is about 61.67% which reflects wide margin of error in the experimentation. The error could have arose from an inconsistent ambient temperature which will affect the Keq significantly. Another could be that the cuvette was not properly rinsed every after reading, leaving some specimen of the previous sample which would then contribute to an aberration in the readings or that proper handling was not executed. Another case would be that the amount of sample placed in the cuvette was either too much or too less than the required amount. The inaccuracy might have also arisen from the preparation of the stock solutions and the test solutions itself. Lastly, the dilution of the test samples or the purity of the reagents used might have been not enough to obtain precise readings. CONCLUSIONS AND RECOMMENDATIONS This experiment was executed with the Sequoia-Turner Model 340 Spectrophotometer and used the BeerLambert-Bougert law in order to obtain a calibration curve which was solved out of the absorbance readings of standard solutions. After using line regression analysis, the calibration solved had the equation, y = 1240.1x - 0.0012, and had an acceptable linearity of R2 = 0.9926, which does not deviate far from ideal values. Though, when the equilibrium constant, Keq, was solved from the equilibrium concentrations of the reactants and product of the unknown solutions, it was found that the experimental Keq = 331.10 was only 66.67% of the literature value for the equilibrium constant of the reaction which is Keq = 890. This experiments final data deviated from the literature value. This could have been contributed by several experimental mistakes such as the reagents used did not have a high enough purity, the solutions were not diluted enough, the ambient temperature where the experiment was done was not consistent, or that the

Initial Change Equim

[Fe3+]init -X [Fe3+]init - X

[SCN-]init -X [SCN-]init- X

0 +X X

Calculated values for the equilibrium concentrations of SCN-, Fe3+, and Fe(SCN)2+ are shown in Table 4. Table 4. Equilibrium Conc. Of Reactants and Product. Soln [Fe(SCN)2+]eq [SCN-]eq [Fe3+]eq -4 -4 Unk 1 1.38 x 10 8.62 x 10 4.66 x 10-4 Unk 2 1.78 x 10-4 8.22 x 10-4 6.22 x 10-4 -4 Unk 3 2.08 x 10 7.92 x 10-4 7.92 x 10-4 Since all pertinent information with regards to the equilibrium concentrations of the unknown has been gathered, the equilibrium constant can then be obtained by using the expression, . From Unknown 1 and so on, the equilibrium constants obtained were 343.55, 348.14, and 331.60, respectively. The average equilibrium constant of the set-up was 341.10.

apparatus, especially the cuvette, were not handled properly and efficiently. This experiment can be improved by using reagents which are of greater purity for the preparation of the stock solutions, or that the solutions to be tested will be more diluted. Lastly, it is recommended that the test solutions must be within the testing room for at least 30 minutes so that the solutions would have similar, if not completely the same temperature, so that the Keq of the solutions would not have drastic differences such as in the experiment. [8, 9] ANSWERS TO QUESTIONS 1. Discuss the significance of the HCl in the solution preparation. The addition of HCl in the solution preparation is done in order to control, i.e., minimize or inhibit, the hydrolysis of the hydrate ferric ion which yields a brown metal-lignin complex, [Fe(H2O)5OH]2+, and/or the brick redcolored rust, Fe(OH)3. If the formation of these colored compounds is not inhibited, the absorbance readings in the experiment would be erroneous since the apparatus will not have absorbance readings that would pertain to the absorbance of the complex of the system alone, Fe(SCN)2+, but would include instead the absorbance of the said brown metal-lignin complex and/or rust. 2. The concentration of [FeSCN]2+ in the standard solution is equal to the concentration of SCN-, the limiting reagent. (Note: In the experiment, CNShas been replaced by Fe3+ as the limiting reagent.) Is this condition always true? If not, what is (are) the condition(s) for this to be true? This will always be true. Since Fe3+ is the limiting reagent, all other additions in excess of other reagents will only push the reaction in the forward direction and will stop once all of the Fe3+ has reacted. All species of FeSCN2+

then must form only from Fe3+ and would have a total concentration equivalent to the moles of Fe3+ added. Moreover, if other complexes form with Fe3+ as its core, the same case will occur. 3. Solutions containing Fe3+ are colored, thus absorb at the visible region. Explain why the absorbance readings in the experiment correspond only to the absorption of the complex, Fe(SCN)2+. This is because colored complexes/ compounds that could arise from Fe3+ have been put in consideration and was dealt with accordingly. two of which are the brown metal-lignin complex Fe(H2O)5OH2+ and/or rust Fe(OH)3 which has been inhibited from formation by acidification of the solutions which prevents the hydrated ferric ion from hydrolyzing. Another is the formation of other complexes such as Fe(SCN)63- which was prevented from developing by limiting the amount of SCN- that was used in the reaction. Hence, the only complex species that was allowed to form was FeSCN2+ whose absorbance was distinctly measured in the experiment. 4. Can distilled water which has zero absorbance be used as blank instead of the Fe3+ solution? This would only contribute to an error since the blank were not used merely for checking the ability of the apparatus to read absorbance data but also used to compensate unnecessary absorbance readings that the test solutions had. This arises from the unreacted amount of SCN- still present in the solutions as well as possible absorbance that the cuvette contributes. It is then from the reading of these unwanted absorbance data that the zero is set on so as to achieve corrected absorbance readings (absorbance wherein the contributed absorbance of miscellaneous parts have been deducted, leaving readings for FeSCN2+ absorbance alone). Otherwise, the readings obtained will be inaccurate since the absorbance value would have

contributions from the absorbance of other parts of the set-up. 5. Account for the difference between the literature value and the experimentally determined value of the equilibrium constant. The error could have arisen from an inconsistent ambient temperature which will affect the Keq significantly. Another could be that the cuvette was not properly rinsed every after reading, leaving some specimen of the previous sample which would then contribute to an aberration in the readings or that proper handling was not executed. Another case would be that the amount of sample placed in the cuvette was either too much or too less than the required amount. The inaccuracy might have also arisen from the preparation of the stock solutions and the test solutions itself. Lastly, the dilution of the test samples or the purity of the reagents used might have been not enough to obtain precise readings. [8, 9]

In marine science, spectrophotometry is also used in finding specific concentrations of dissolved oxygen in the water, salinity, and trace presence of minerals. This is often used in quality assessment of marine and fresh water environments. REFERENCES
[1]Dodd,

R.E. Chemical Spectroscopy. Elsevier Publishing Company. 1925.

[2]R.

W. Ramette, J. Chem. Ed. 40, 71 (1963) F.A., et al. Laboratory Experiments For General, Organic, and Biochemistry, 7th Ed. Brooks/ Cole Cencage Learning. A.E., Hancock, R.D. Metal Complexes in Aqueous Solutions. A Division of Plenum Publishing Corporation. 1996. Douglas C., (2005). PHYSICS Principles with Applications, 6th Ed. Pearson Education, Inc.

[3]Bettelhelm,

[4]Martell,

[5]Giancoli,

[6]F.

APPLICATIONS The spectrophotometric determination of the equilibrium constant has many applications in the different industries and related sciences. It can be applied in the food industry in the quality testing of honey or maple syrup and spectral analysis of fruit juices. In medicine, it is used in the determination of drug behavior in specific and controlled environmental conditions and shifting equilibrium concentrations (e.g. Treatment for sickle-cell anemia and its effect on oxygen transport) as well as in the determination of cholesterol amounts in clinical samples. This process is also used for the measurement of the characteristics of paint, varnish, and lacquer, which include, but may not be limited to color, specular and contrast gloss, bloom and sheen.
[7]H.

A. Cotton and G. Wilkinson, Advanced Inorganic Chemistry, Interscience, 1962, p. 717. S. Frank and R. L. Oswalt, J. Am. Chem. Soc. 69, 1321 (1947) C.L., Love, G.A. Journal of Chemical Education. Volume 75, Number 1.

[8]Cobb,

[9]C.

L. Cobb and G. A. Love, J. Chem. Ed. 75, 90 (1998) J. Stolzberg, J. Chem. Ed. 76, 640 (1999) R.H., et al. General Chemistry, 10th Ed . 2010. J.A. Chemical Principles in Practice. Addison-Wesley Publishing Company. 1967.

[10]R.

[11]Petrucci,

[12]Bell,

CALCULATIONS A. Calibration Curve

M1V1 = M2V2 M2 = [Fe3+]init = [Fe3+]stock[V]stock/[V]tot Std 1 [Fe3+]init = 0.002 M (0.1 mL)/ 10 mL = 2.0 x 10-5 M Std 2 [Fe3+]init =0.002 M (0.25mL)/ 10mL = 5.0 x 10-5 M 3+] Std 3 [Fe init = 0.002 M (0.5 mL)/ 10 mL = 1.0 x 10-4 M Std 4 [Fe3+]init = 0.002 M (1.0mL)/ 10 mL = 2.0 x 10-4 M 3+] Std 5 [Fe init = 0.002 M (2.0 mL)/ 10 mL = 4.0 x 10-4 M Note: Since Fe3+ is the limiting reagent, [FeSCN2+]eq will have the same concentration for its corresponding [Fe3+]. [SCN-]init = [SCN-]stock[V]stock/[V]tot Std 1 [SCN-]init = 0.2 M (1.0 mL)/ 10 mL = 0.02 M Note: Std 2 to Std5 has the same computation and answer. B. Absorbance of Unknown Solutions M1V1 = M2V2 M2 = [Fe3+]init = [Fe3+]stock[V]stock/[V]tot

Unk 1 [FeSCN2+] = = 1.38 x 10-4 M Unk 2 [FeSCN2+] = = 1.78 x 10-4 M Unk 3 [FeSCN2+] = = 2.08 x 10-4 M

[Fe3+]

[SCN-]

[Fe(SCN)2+]

Initial Change Equim

[Fe3+]init -X [Fe3+]init - X

[SCN-]init -X [SCN-]init- X

0 +X X

X = [FeSCN2+]eq
[Fe3+]eq = [Fe3+]init X [SCN-] eq= [SCN-]init- X

= [Fe3+]init [FeSCN2+]eq = [SCN-]init- [FeSCN2+]eq

Unk 1 [Fe3+]eq = 6.0 x 10-4M 1.38 x 10-4M

= 4.66 x 10-4 M Unk 1 [Fe3+]eq = 8.0 x 10-4M 1.78 x 10-4M = 6.22 x 10-4 M Unk 1 [Fe3+]eq = 1.0 x 10-3M 2.08 x 10-4 = 7.92 x 10-4 M

. Unk 1 Keq, 1 = = 343.55 Unk 1 Keq, 2 = = 348.14 Unk 1 Keq, 3 = = 331.60

Unk 1 [Fe3+]init = 0.002 M (3.0mL)/ 10mL = 6.0 x 10-4 M 3+]init = 0.002 M (4.0mL)/ 10mL Unk 2 [Fe = 8.00 x 10-4 M 3+]init = 0.002 M (2.0mL)/ 10mL Unk 3 [Fe = 1.0 x 10-3 M C. Equilibrium Constant Determination , Keq, Calibration Curve A = bc y = 1240.08x 0.0012 where y = A (Absorbance) b = 1 cm = 1240.08 c = [ FeSCN2+]eq = x x= =c

Keq,AVE = = = 341.10

%error = | = | = 66.67% | x 100%

| x 100%