Lab 3- Spectrophotometric Analysis; 10/7/2011

Rahul Goel SID- 23077974

ABSTRACT In this lab, various samples of Chromium, Cobalt and Manganese were analyzed with the help of spectrophotometric analysis, statistical analysis and BeerLambert Law. We obtain the absorbance of different samples at various concentrations using the spectrophotometer. And then by solving various systems derived from the beers law and finding the molar extinction coefficient and putting it in the equations of samples having unknown concentration, we determine the concentration of various unknown samples. Initially, we were given some solutions of known and unknown concentrations from the stock room. The solutions of known concentrations were diluted to various lower known concentrations. Then, the samples of various concentrations are filled in cuvets and set in the spectrophotometer beginning the analysis with water as a reagent blank whose absorbance gives us an idea of the baseline and the starting point for all our calculations. And then, all the calculations are done. In our experiments, the concentrations of manganese were found to be 0.17469 mmolar, 0.21562 mmolar and 0.25853 mmolar for Samples 1, 2 and 3 respectively, while the concentrations of Chromium and Cobalt were found to be 7.9440 mmolar and 32.218 mmolar respectively. INTRODUCTION Spectrophotometry based on Beer-Lambert Law is one of the most convenient and widely used analytical methods. The device we use for this is known as spectrophotometer or a spectrometer and it consists of two main parts- a spectrograph which produces light of selected wavelengths and is calibrated to indicate the color of the light it produces in terms of wavelength in nanometers(usually) AND a photometer which measures intensity of the given beam at a wavelength or in other words tells us about the absorbance of a given

Lab 3- Spectrophotometric Analysis; 10/7/2011

sample at a particular wavelength. Absorbance or the amount of light absorbed by the sample is used to determine the amount of colored component in the solution according to the Beers Law. BEER-LAMBERT LAW In the determination of the concentration of manganese in the first part of lab, we use the basic beers law equation. A = bc Here, A is the absorption of the solution containing the given solution. A is the equal to the product of molar absorptivity times the path length which the light has to travel b times the concentration c. We have particular units for all the above mentioned physical quantities. is measured in liters per mole-cm and concentration c is measured in moles or millimoles per liter depending on the provided sample. Talking about the path length b, the special sample cuvets have b=1.0000 cm and we also used the same type of cuvets in our experiments. For the second part of the experiment, a system of equations derived basically from the beers law is used. Here, the basic principle involved is the fact that the total absorbance of a solution at a given wavelength is equal to the sum of the wavelengths of the individual components. So, we can analyze the individual components of a mixture even their spectra overlap. In our experiment, the two components were Cobalt(II) and Chromium(III) and the absorbance of solutions of both the elements are measured for various concentrations at two wavelengths, which are chosen such that the difference in molar absorptivities between the two ions is large at these wavelengths. So, the absorbances of mixture at the two wavelengths 1 and 2 can be expressed as: A1 = M1 bc M + N1 bc N A2 = M2 bc M + N2 bc N Solving the above system of equations simultaneous, we can find the concentration of individual components.

Lab 3- Spectrophotometric Analysis; 10/7/2011

Determination of the concentration of Manganese in an unknown sample Manganese is a transition element and forms intensely colored (purple) permanganate ion in its +7 oxidation state. Manganese is added to steel to enhance its toughness. Steel is an alloy containing mostly iron, manganese, copper and other metals. We use 2 steps to determine the unknown concentration. Firstly, we do the spectrophotometric of the known standard solutions. The absorbances of samples at known concentrations are analyzed by spectroscopy. Ultraviolet-visible spectroscopy measures the amount of energy a molecule absorbs at a certain wavelength. And absorbance of a sample is based on the amount of light that passes through a sample by Beers Law. We measured the absorbance at a wavelength of 526 nm, reason being that this wavelength absorbs yellow-green light which is the light color absorbed by the magnesium sample. Initially, the concentration of the sample was .29242 millimolar. In this experiment, the unknown sample had Mn2+ ions, and then these ions were oxidized to MnO4- ions with potassium periodate (KIO4). Manganese ions are purple in color and their absorbance can be measured by ultraviolet-visible spectroscopy. Phosphoric acid was added to the solutions to increase the rate of oxidation and to prevent the precipitation of manganese(II) periodate or iodate. The reaction equation is: 2Mn2+ + 5IO4- + 3H2O 5IO3- + 6H+ + 2Mn(VII)O4For part 2 of this experiment, we again prepare diluted solutions from the given solutions of chromium (0.05M) and cobalt (0.2M). And then again, the samples are analyzed by the spectrophotometer and the absorbances for various samples are noticed at wavelengths 407nm and 511nm. These wavelengths are chosen in such a manner that the differences between molar absorptivities are large between the two ions. From the data on absorbances, the calibration curves are plotted for both wavelengths of chromium and cobalt and their slopes (b) are

Lab 3- Spectrophotometric Analysis; 10/7/2011

determined. Also, their 95% confidence level is determined, assuming that all the uncertainty comes from the slope measurements. Using this data, we calculate the concentrations of the unknown solutions.

EXPERIMENTAL METHODS Since this experiment involved many steps of dilution, it is critical to use clean and dry containers, so that one gets uncontaminated solutions with accurate concentrations. Also all the glassware must be scrupulously clean of dust and soap so the concentrations of solutions do not get affected (for example, permanganate concentration changes easily with time as the organic material gets oxidized). Solutions were prepared by diluting the original solutions of concentrations .29242 mM MnO4- in Part A, 0.05M Cr(III) and 0.2M Co(II) in Part B. For all of dilution process, small amount of original/previously prepared solutions were filled into volumetric flasks and distilled water was added up to 50mL mark. For example, let us discuss the dilution of Chromium solution. Initially we took 50 ml .05 M solution. To get 0.04 molar, we took 40 ml 0.05 molar solution + added 10 ml of water to it. For 0.03 molar, take 30 ml 0.05 molar solution and add 20 ml water. For 0.02 molar solution, take 25 ml of 0.04 M solution and add 25 ml of water. And finally for 0.01 molar solution take 25 ml of 0.02 molar solution and add 25 ml of water. It is important to carry out dilutions carefully so the concentrations are as close as possible to the desired concentrations. Also the solutions should be thoroughly mixed after each dilution. Spectrophotometer was used to measure the absorbance of each solution transferred into cuvettes, and specific wavelengths were used for Part B so that they produce the greatest accuracy. In Part A, three peak wavelengths were chosen for the same purpose, and in this experiment, spectrophotometer was set up so that it would show the absorbance at the chosen wavelengths. RESULTS Symbols necessary to know: m=slope; sm=std dev of m; sy=std dev of absorbance

Lab 3- Spectrophotometric Analysis; 10/7/2011

LINEST m=1.551371684 sm=0.105073981 R =0.981981365

2

b=0 sb=#N/A sy=0.045178

Mean Std dev N Deg. of freedom Conf. level t value Conf. interval

0.252175 0.183811 5 4 0.95 2.776445 0.228232

Potassium Permanganate at wavelength526 nm

0.6
0.5 0.4 0.3 0.2 0.1 0 0.1052712, 0.117796 0.175452, 0.216303 y = 1.551371684x 0.233936, 0.37851 0.29242, 0.49648

Absorbance

0.0526356, 0.051785

0.05

0.1

0.15

0.2

0.25

0.3

0.35

Concentration (mmolar)

PLOTS FOR CHROMIUM LINEST m=8.562254545 sm=1.494551387 R2=0.891366954 b=0 sb=#N/A sy=0.110839

Lab 3- Spectrophotometric Analysis; 10/7/2011

Mean Std dev N Deg. of freedom Conf. level t value Conf. interval 0.298984 0.036746 5 4 0.95 2.776445 0.045626 0.45 0.4

Chromium(III) at wavelength-511 nm
y = 8.5623x 0.310094 0.28489 0.25448 0.29134 0.35412

0.35
0.3 Absorbance 0.25 0.2

0.15 0.1 0.05 0 0 0.01 0.02 0.03 0.04 Concentration (Molar) 0.05 0.06

LINEST m=13.2446 sm=1.488664188 R2=0.951897677 b=0 sb=#N/A sy=0.110402

Mean Std dev N Deg. of freedom Conf. level t value Conf. interval

0.439332 0.110112 5 4 0.95 2.776445 0.136722

Lab 3- Spectrophotometric Analysis; 10/7/2011

0.7 0.6 0.5 Absorbance

Chromium(III) at wavelength- 407 nm

y = 13.245x 0.586794 0.499054

0.4 0.3 0.30046 0.2 0.1 0 0.37716

0.433199

0.01

0.02 Concentration (Molar) 0.04 0.03

0.05

0.06

PLOTS FOR COBALT

Mean Std dev N Deg. of freedom Conf. level t value Conf. interval

0.187026 0.166209 5 4 0.95 2.776445 0.206376

m=2.264335612 sm=0.038142883

b=0 sb=#N/A

R2=0.998866262 sy=0.008994

Cobalt(II) at wavelength- 511 nm

0.5 Absorbance 0.46213

0.4
0.3 0.2 0.1 0 0 0.12741 0.083851 0.046271 0.05 0.1

y = 2.2643x 0.21547

0.15

0.2

0.25

Concentration (Molar)

Lab 3- Spectrophotometric Analysis; 10/7/2011

LINEST m=0.221274428 sm=0.002958498 R2=0.999285455 b=0 sb=#N/A sy=0.000698

Mean Std dev N Deg. of freedom Conf. level t value Conf. interval

0.018287 0.016219 5 4 0.95 2.776445 0.020138

Cobalt(II) at wavelength- 407 nm

0.05 0.045 y = 0.2213x

0.04492

0.04
0.035 Absorbance 0.03 0.025 0.021591 0.012493 0.0080781 0.0043527 0 0.05 0.1 0.15 Concentration (Molar) 0.2 0.25

0.02
0.015 0.01 0.005

DISCUSSION In this laboratory experiment, we learnt to use the spectrophotometer for analysis of various samples. A strong emphasis was laid on proper dilution techniques so that proper solutions with right concentrations are obtained and better results are obtained. We learnt that the spectrophotometer can be used to find the concentration of any unknown sample if the absorbance of the sample of known concentration is available. We use a combination of Beers Law and statistics to analyze the unknown concentration.

Lab 3- Spectrophotometric Analysis; 10/7/2011

The final results are discussed now. For manganese samples, the concentrations were 0.17469 mmolar, 0.21562 mmolar and 0.25853 mmolar for Samples 1, sample 2 and sample 3 respectively, while the concentrations of Chromium and Cobalt were found to be 7.9440 mmolar and 32.218 mmolar respectively. Talking in particular about the chromium(III) samples, the plot for values achieved at wavelengths 407 nm and 511 nm were nearly linear. But, a strong problem observed was that the graph did not pass through the origin and was of the form y=mx+c instead of y=mx. To correct this, a trendline passing through the origin was chosen. Though it is quite off the curve, but this experimental error could be fixed by only this method. There can be several reasons for these errors. Firstly, dilutions were done using volumetric flasks and pipets which are note that accurate and some error creeps in while measuring as it is difficult to measure the exact amount by pipetting. Secondly, the same cuvets were used again and again during the experiment. Though they were emptied, but some drops of the previous sample could still be sticking to the walls of the cuvets. Even if we rinse the cuvets before pouring in a fresh sample, the presence of water droplets in the cuvets could decrease the concentration. While the presence of the sample drops on the walls can both increase or decrease the concentration of the current sample poured. Moreover, the containers might have been contaminated and not properly rinsed. It can affect the results drastically by changing the concentration or even the composition of the sample. Also, one might note down the wrong value of baseline and results would shifts up or down by a y intercept. One other reason can be that the water used in the experiment is not distilled and may contain some ions in it. Finally, the same spectrophotometer does not give the same values and gives different results for the same sample each time we measure the absorbance at a particular wavelength. Talking about the calculations and interpretation of the data, it was done using the Beer- Lambert Law. In the determination of manganese, the simple formula A= bc was used. While for the mixture of the cobalt and chromium, a more complicated set of equations based on the beers law was used.

Lab 3- Spectrophotometric Analysis; 10/7/2011

The sources of error can be corrected in a number of ways. First of all, we may note down the wrong baseline for our calculations. So, it needs to be checked properly. Moreover, all the apparatus must be clean and dry so that there is no contamination and hence no change in concentration or composition of sample is there. Also, the dilutions must be done carefully especially focusing on pipetting the right amount of sample as even one drop extra or less can change the entire concentration of the sample. In a nutshell, one should be more careful and focused so as to get more accurate results. RESOURCES Harris, Daniel C. Qualitative Chemical Analysis ; Eighth Edition; W.H. Freeman and Company, New York, 2010 Laboratory Manual, Chemistry 4A Fall 2011; Department of Chemistry, University of California, Berkeley