APPENDICES A. Data Sheet B.

Sample Calculations WORKING EQUATIONS Initial Concentrations of the Reagents

M initial =

M 1V1 Vtotal

Beer-Lamberts Law

A= L C
where A absorbance molar absorptivity b path length c concentration of absorbing species

Equilibrium Expression for the Reaction

K eq =

[[ Fe ( SCN )] 2 ] [ Fe 3 ][ SCN ]

Determination of [[Fe(SCN)]2+] in Unknown Solutions

C=

% Difference | |

For the standard solutions: [SCN-] = mmol SCNtotal volume solution [Fe(SCN)]2+ = [SCN-] m (slope) = b b = 1 cm = m/b

For the unknown solutions: [SCN-]init = mmol SCNtotal volume solution [Fe3+]init = mmol Fe3+ total volume solution [Fe(SCN)]2+eq = A 0.0135 3327.5 [SCN-]eq = [SCN-]init - [Fe(SCN)]2+eq [Fe3+]eq = [Fe3+]init - [Fe(SCN)]2+eq Keq = [Fe(SCN)]2+eq [SCN-]eq [Fe3+]eq

% Difference = |Keq exp - Keq theo| x 100 Keq theo

SAMPLE CALCULATIONS
A. Equilibrium Concentration of Fe(SCN)2+ in Standard Solutions Standard 1: 0.20 ml SCN- (

)(

= 0.00004 M SCN- at equilibrium =[Fe(SCN)]2+ at equilibrium [Fe(SCN)]2+ (eq) = 0.00004 M

B. Equilibrium Concentration of Fe(SCN)2+ in Unknown Solutions Equation of Line(Absorbance vs [FeSCN]2+ (eq)): A = 2205x + 0.061 Unknown 1: A = 2205x + 0.061 0.071= 2205x + 0.061 0.071-0.061 = 2205x 2205x = 0.01 X = 0.000004535 M or

C. Initial Concentrations of Fe3+ and SCNMolar Concentration of FeCl3: 0.002 M Concentration of KSCN: 0.002 M Unknown 1: ( ( )( )( ) )

D. Equilibrium Concentrations of Fe3+ and SCN- in Unknown Solutions Equilibrium Concentration = Initial Concentration of Reactant Equilibrium Concentration of Product Unknown 1: Fe3+(eq )= 0.001 M 0.000004535 M = 0.000995 M or 9.95 x 10-4 M SCN-(eq) = 0.0002 M 0.000004535 M = 0.000195 M or 1.95 x 10-4 M E. Keq of Unknown Solutions

( [ ]

Unknown 1:
( )( )

Keq = 23.4

F.

% Difference

Keq average: 112 Literature Value: 920

| | = 87.8 % A. SAMPLE CALCULATIONS [Fe(SCN)]2+ = [SCN-] | x 100

| x 100

[SCN-] = (0.20 mL)(0.002 M) = 4.0 x 10-5 M 10.00 mL [SCN-] = (0.40 mL)(0.002 M) = 8.0 x 10-5 M 10.00 mL [SCN-] = (0.60 mL)(0.002 M) = 1.2 x 10-4 M 10.00 mL [SCN-] = (0.80 mL)(0.002 M) = 1.6 x 10-4 M 10.00 mL

= 3,327.5 cm-1M-1 = 3,327.5 M-1 1 cm

[Fe3+]init = (5.00 mL)(0.002 M) = 0.00100 M 10.0 mL [SCN-]init = (1.00mL)(0.002M) = 2.00 x 10-4 M 10.0 mL [SCN-]init = (2.00mL)(0.002M) = 4.00 x 10-4 M 10.0 mL [SCN-]init = (3.00mL)(0.002M) = 6.00 x 10-4 M 10.0 mL [SCN-]init = (4.00mL)(0.002M) = 8.00 x 10-4 M 10.0 mL [SCN-]init = (5.00mL)(0.002M) = 1.00 x 10-3 M 10.0 mL

[Fe(SCN)]2+eq = 0.158 0.0135 = 4.34 x 10-5 M 3327.5 [Fe(SCN)]2+eq = 0.308 0.0135 = 8.85 x 10-5 M 3327.5 [Fe(SCN)]2+eq = 0.457 0.0135 = 1.33 x 10-4 M 3327.5 [Fe(SCN)]2+eq = 0.604 0.0135 = 1.77 x 10-4 M 3327.5 [Fe(SCN)]2+eq = 0.743 0.0135 = 2.19 x 10-4 M 3327.5

[SCN-]eq = [SCN-]init - [Fe(SCN)]2+eq = 2.00 x 10-4 M 4.34 x 10-5 M = 1.57 x 10-4 M [SCN-]eq = [SCN-]init - [Fe(SCN)]2+eq

= 4.00 x 10-4 M 8.85 x 10-5 M = 3.12 x 10-4 M [SCN-]eq = [SCN-]init - [Fe(SCN)]2+eq = 6.00 x 10-4 M 1.33 x 10-4 M = 4.67 x 10-4 M [SCN-]eq = [SCN-]init - [Fe(SCN)]2+eq = 8.00 x 10-4 M 1.77 x 10-4 M = 6.23 x 10-4 M [SCN-]eq = [SCN-]init - [Fe(SCN)]2+eq = 1.00 x 10-3 M 2.19 x 10-4 M = 7.81 x 10-4 M

[Fe3+]eq = [Fe3+]init - [Fe(SCN)]2+eq = 0.00100 M 4.34 x 10-5 M = 9.57 x 10-4 M [Fe3+]eq = [Fe3+]init - [Fe(SCN)]2+eq = 0.00100 M 8.85 x 10-5 M = 9.12 x 10-4 M [Fe3+]eq = [Fe3+]init - [Fe(SCN)]2+eq = 0.00100 M 1.33 x 10-4 M = 8.87 x 10-4 M [Fe3+]eq = [Fe3+]init - [Fe(SCN)]2+eq = 0.00100 M 1.77 x 10-4 M = 8.23 x 10-4 M [Fe3+]eq = [Fe3+]init - [Fe(SCN)]2+eq = 0.00100 M 2.19 x 10-4 M = 7.81 x 10-4 M

Keq = [Fe(SCN)]2+eq [SCN-]eq [Fe3+]eq = 288.85 Keq = [Fe(SCN)]2+eq [SCN-]eq [Fe3+]eq = 311.02 Keq = [Fe(SCN)]2+eq [SCN-]eq [Fe3+]eq = 321.08 Keq = [Fe(SCN)]2+eq [SCN-]eq [Fe3+]eq = 345.21 Keq = [Fe(SCN)]2+eq [SCN-]eq [Fe3+]eq = 359.04 Average Keq value = 325.04

Theoretical Keq value of [Fe(SCN)]2+ = 890

% Difference = |Keq exp - Keq theo| x 100 Keq theo = |325.04 - 890| x 100 890 = 63.5%

C. Answers to Questions 1. Discuss the significance of the HCl in the solution preparation. The HCl dilutes the solutions in the system used because Beers law, which relates the absorbance to the concentration only applies to dilute solutions.

2. The concentration of FeSCN2+ in the standard solution is equal to the concentration of CNS-, the limiting reactant. Is this condition always true? If not, what is(are) the condition(s) for this to be true? A blank solution was used to cancel out absorption by any other solutes, so that the absorbance values that will be reported are directly proportional only to the solute of interest, which, in this experiment, is the FeSCN2+. Since the blank solution does not contain any FeSCN2+, and contains HCl and FeCl3, the spectrophotometeric readings will correspond only to the absorbance of FeSCN2+. Since the only equilibrium concentration that we wanted to measure is that of FeSCN2+, diluted FeCl3 in hydrochloric acid should really be used; but if distilled water was used as the blank, zero absorbance would be recorded for the blank solution. But the absorbance of the Standard Solutions that would be reported is the absorbance of [Fe(SCN)]2+, HCl and Fe3+ (Fe3+ is an absorbing species), so we cannot obtain the absorbance of [Fe(SCN)]2+. The concentration of FeSCN2+ in the standard solution is only equal to the limiting reagent, SCN-, if the volume or amount of thiocyanate ions is very small, where it is the definite limiting reagent and there is a large excess Fe3+. This condition will drive the reaction to the right, consuming all thiocyanate ions yielding FeSCN2+, therefore the equilibrium concentration of FeSCN2+ equals the initial concentration of SCN-. For the unknown solutions, the given statement does not follow. It appears that there must be a greater number of moles of Fe3+ in comparison to the number of SCN- moles in order for the given statement to hold true. Hence, this condition is not always true. A condition would be that the limiting reagent, thiocyanate, should be very small compared to the excess reagent. This is because, if the limiting reagent is very small compared to the excess reagent, the excess reagent would push the reaction forward, consuming the limiting reagent. 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, FeSCN2+. Beers law applies to only one absorbing solute in the solution. A blank solution was used to cancel out absorption by any other solutes, so that the absorbance values that will be reported are directly proportional only to the solute of interest, which, in this experiment, is the FeSCN2+. Since the blank solution does not contain any FeSCN2+, and contains HNO3 and Fe(NO3)3, the spectrophotometer readings will correspond only to the absorbance of FeSCN2+. Another thing is that nitric acid is added into the solution. The acid clarifies the solution, minimizing the yellowish-brown color of Fe3+, making the Fe(SCN)2+ the predominantly absorbing species. Although solutions containing Fe3+ absorb at the visible region, this does not mean that it also absorbs strongly at the ultraviolet region. Therefore, it is not a completely absorbing species. SCN- acts as a chromophoric reagent to produce [Fe(SCN)]2+, which strongly absorbs in the ultraviolet and visible regions. Since [Fe(SCN)]2+ is the absorbing species in the experiment, the absorbance readings will correspond to its absorption capacity.

4. Can distilled water which has zero absorbance be used as blank instead of the Fe3+ solution? Since the only equilibrium concentration that was to be measured was that of FeSCN2+, diluted Fe(NO3)3 in nitric acid should really be used but if distilled water was used as the blank, zero absorbance would be recorded for the blank solution, but the absorbance of the Standard Solutions that would be reported is the absorbance of [Fe(SCN)]2+, HNO3 and Fe3+ (Fe3+ is an absorbing species),

so one cannot obtain the absorbance of [Fe(SCN)]2+. Although both samples (distilled water and diluted Fe(NO3)3 in HNO3) have both zero absorbance readings, the goal is to cover the overall composition of the samples and to encompass a wider range of the concentrations of the absorbing species. Distilled water does not tell anything related to the overall composition of the samples. 5. Account for the difference between the literature value and the experimentally determined value of the equilibrium constant. In the experiment, several instrumental deviations may have been encountered which might have lead to differences in the absorbance readings. Some of these are mismatched cells and stray light in the spectrophotometer. These deviations lead to errors in the experimental value of the molar absorptivity. When molar absorptivity is used from the calibration curve later on for the computation of the equilibrium concentrations of [Fe(SCN)]2+, Fe3+, and SCN, the error is propagated and is even more developed upon the computation of the equilibrium constant. Another possible source of this difference is that the reaction of the Fe3+ with the thiocyanate is incomplete and less product is produced, which means there is less absorbing species in the sample leading to a lower absorbance reading. The computed average equilibrium constant for the formation of [Fe(SCN)]2+ was 104.56 which differs by 51.9% from the literature value which is 1047.13. The mentioned errors were possibly the cause of the difference between the experimental and literature values. If these errors were carefully avoided, the margin between the two values, theoretical and experimental, would be narrowed.

APPLICATION(S)

Spectrophotometry is a special type of spectroscopy that has a lot of uses in the field of Physical Chemistry. It can be used in the determination of glucose levels. A study conducted to determine the glucose concentrations in the blood of diabetic patients by the Biochrom Clinics uses the physiological fluids in the cuvette directly and calculate its glucose concentration.

The use of the spectrophotometer has also applications in trade. They use spectrophotometry to determine the authenticity of the fruit juices especially orange juice. The Florida Department of Citrus devised a method in knowing whether the citrus products are authentic or not. They place a sample in the cuvette and scans for vitamin c which has a wavelength peak of 250nm and the absorbance ratio must be 443/325nm. If the product is a fake and contains flavonoid, the sample peaks at 280nm. Also, to further study the samples the concentration of vitamin c was obtained to check if it matches the required level. Hospitals have also devised a way to determine the protein metabolism index of a person to be used in their nutritional and health status. One the product of protein metabolism is urea. In the procedure urea is hydrolised to ammonia and carbon dioxide. Ammonia reacts with axoglutarate then the product is put in the spectrophotometer at 340nm. If the resulting products concentration is in the normal range the person has a normal protein metabolism. Spectrophotometry and photometric methods have many important characteristics that make it one of the most useful tools for quantitative analysis. It has a very wide applicability because it can be used on both absorbing and nonabsorbing species. The detection limits for absorption spectroscopy can also be extended to 10-6 or even 10-7 M indicating its high sensitivity. It also offers moderate to high selectivity since specific wavelengths can be found at which the analyte alone absorbs so there in no need for a separation step. It is also a very accurate method since the range of errors in the procedure lie only with 1% - 5% and can even be decreased with special precautions. With modern instruments, spectrophotometric and photometric measurements can easily and rapidly be performed. Non-absorbing species can be determined after chemical conversion into absorbing derivatives. These species are caused to react with chromophoric reagents to give products that strongly absorb in the ultraviolet and visible regions. Usually, the reaction of the color-forming reagents with the analyte is forced near completion for the successful application of the reagents. There is a large number of inorganic, organic, and biochemical species that absorb ultraviolet or visible radiation and can be determined through direct quantitative methods. Ions of the transition metals are colored in solution and can thus be identified through spectrophotometric measurement. Ultraviolet and visible absorption spectroscopy account for more than 90% of the analyses performed in clinical laboratories.