CHEM 111L Lecture Notes - Lecture 6: Scatter Plot, Food Coloring, Molar Attenuation Coefficient

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kurtzkm

17 Apr 2019

School

College of Charleston

Department

Chemistry and Biochemistry

Course

CHEM 111L

Professor

Guirgis

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Document Summary

Experiment 6 - absorbance of light vs concentration. Determine the concentration of fd&c dyes in known and unknown solutions. Understand the relationship of absorbance of light and color. A: ~20 ml of original sample of food dye. B: 5. 00 ml of dye + 5. 00 ml of di. C: 2. 00 ml of dye + 8. 00 ml of di. D: 1. 00 ml of dye + 9. 00 ml of di. Measure absorbance of other 3 solutions with the same wavelength as d. Enter concentration in column a and absorption in column b. Add a linear trendline with options shown in photo: Collect a sample of a commercial drink and write down the unknown number. Measure the absorbance of the solution at the max wavelength for each of the dyes. Use the beer"s plot for each dye to determine the concentration of the dye in the drink. A is the amount of light energy absorbed. Is a constant called the molar absorptivity.

Related Questions

Topic: spectroscopy: examination of the absosorption spectrum and determination of the extinction coefficient for Flavin mononucleotide

2) in part 2 of the experiment, you are asked to prepare 5.0 mL of 2.0 x10-5M FMN starting with a 1.0 x 10 -4 solution of FMN? Calculate the amounts of 1.0 x 10-4 M FMN solution and water needed.

ice POT Il. Experimental Procedure 1) Examination of the absorption spectrum of FAIN. Prepare 5.0 mL of 1 x 10 4 MI FMN by luting 0 10 ml of 5.0 x 103 M FMN into 4.90 mL of water. Place about 3 mL in a plastic cuvette and take absorbance readings every 10 nm between 340 nm and 600 nm. (Use water as the blank and be sure to re-zero the blank every time you change the wavelength.) Tabulate the absorbance readings in Table l. Inspect the data and determine where the absorbance is maximal for use in the next part. Plot absorbance (y) vs. wavelength (x) in Excel. Be sure to label the axes correctly 2) Prepare 5 0 mL of 2.0 x 10-5 M FMN by diluting the 1.0 x 10-4 M FMN from part #1 appropriately. Place part of the new FMN sample you prepared in a quartz cuvette provided by the TA Collect an absorbance spectrum of the sample between 190 nm and 1100 nm using the Shimadzu UV1800. (Use water as your blank in the reference cuvette). Export your data to a thumb drive for later analysis in Excel. (Note- everyone in the lab must share the 4 Shimadzu spectrophotometers, so you may need to wait for your turn to complete this part. If so, proceed to parts 3 and 4 and return to this when one of the instruments is free.) 3) Construction of a calilbration curve. Prepare 4.0 mls of 5x104 M FMN by diluting the 5 x103 M FMN appropriately. Prepare 6 test tubes as indicated in the table below. Calculate the concentration of FMN present in each of the 6 tubes and enter concentrations in Table II. Set the wavelength on the Genesys to the wavelength where the absorbance of FMN is maximal in the visible region and measure the absorbance of each of the 6 test tubes. Record these values in Table II. (At the wavelength where the absorption is maximal we will get the greatest response and inaccuracies in setting the monochromator will be the smallest, since the slope of the absorbance spectrum at this point is the least.) Plot the absorbance vs, concentration in Excel. In Excel, add a linear regression trendline that goes through (0,0) and determine the slope of the line. This slope gives the extinction coefficient for the FMN under these conditions. Tube # 6 mL of 5x 104 M FMN0.1 02 04 0.6 0.8 1.0 4.9 48 4.644 42 4.0 mL of water 4) Determination of unknown. Dilute 1.0 mL of the FMN solution with unknown concentration into 4.0 ml of water. Measure the absorbance and record it in Table IlI Repeat this three times and average your results

Spectrophotometric Determination of Iron in a Vitamin Pill Introduction Iron is one of the essential elements in the human diet. In the body, iron is present in the form of dietary iron results in the condition of anemia. Many people take vitamin pills containing iron to supplement their dietary intake ofiron. The technique of is method of instrumental analysis. Many different types spectrophotometers are available that a of the electromagnetic radiation transmit light in different regions spectrum. In is experiment, a visible spectrophotometer will be used. A visible spectrophotometer of a measures the amount of light that a colored solution absorbs. When a beam o the sample depends on wavelength passes through a colored solution, the amount of light absorbed by the concentration of what the substance is that is absorbing the light. The absorbance also depends on the the path through the light passes. The relationship between absorbance, concentration, and pathlength is expressed by the equation known as Beers la gte where A is the al of the solution, E is a constant called absorptivity, which is a characteristic of the substance and the wavelength of the absorption, c is the concentration of the absorbing substance in is a unitless the solution, and e is the length of the path through which the light passes. Abe quantity, concentration can be expressed in any convenient concentration unit, and the pathlength is measured in centimeters. The units of absorptivity depend on the concentration units used. In this experiment concentration is measured in mg/mL Therefore, the units of e are mL/mg cm To determine the absorptivity of a particular substance, a calibration curve is constructed. A series of standard solutions in which the concentration of the absorbing species is known are prepared and their absorbances absorbance as a function concentration is made. If the absorbin species obeys Beer's law, the plot will be a straight line, y mr b. When the pathlength is 1.0 cm, the slope of the line is the absorptivity. The absorbance of a solution of unknown concentration can be read and its concentration determined from the calibration curve. In this experiment, a commercial vitamin pill will be analyzed for its iron content. The form of iron present in the vitamin pill, most likely iron(T) sulfate, has a negligible visible absorbance. The iron(T) will be converted to a species that has a large absorbance, allowing the absorbances of solutions of low iron(T) concentration to be measured. The reagent 1.10-phenanthroline, Figure 1, reacts with is red-orange and absorbs visible ligh most strongly at 508 nm. Iron(ll) is easily oxidized to iron(TI) in aqueous solution. A reducing agent, hydroquinone, is added to the solution to prevent this oxidation from occurring. In addition, the acidity of the solution must be controlled to ensure the formation of the complex. In this experiment, sodium citrate is added to maintain the proper pH of the solution A series of standard solutions with known iron concentrations is prepared, the absorbances of the solutions are read, and a calibration curve constructed. From the calibration curve, the concentration of iron in a solution made from the dissolved vitamin pill will be determined and the total amount of iron in the vitamin pill calculated.

THE ONE IN ITALICS AND BOLD IS THE PRELAB QUESTION PLEASE ANSWER THAT. PLEASE HELP ME ANSWER THE POST LAB QUESTIONS. I did THE PRE LAB. I ALSO FILLED in the tables.

Determination of Cobalt (II) Chloride

by UV/VIS Spectroscopy

Introduction

The absorption of specific quantities of electromagnetic (light) radiation by an element or compound allows electrons to move from lower energy level to a higher energy level. We say that the energy levels are quantized. Electrons, returning to lower energy levels, will release energy within the electromagnetic spectrum. The wavelengths may be in the ultraviolet, visible, or infra red regions of the spectrum. For cobalt (II) chloride, the compound of study, the visible region, of the spectrum will be addressed.

By knowing the wavelength (l) or frequency (n) of radiation, one may determine the energy of the transition. Relationship between energy, wavelength and frequency are:

c= ln and E= hn or E= hc/l

The light radiation absorbed in a solution of this salt is proportional to its concentration through a relationship provided by the Beer-Lambert Law:

A = kC

where A= absorbance, k is Beerâs Law constant, and C is the molar concentration in solution.

The absorbance (A), a unit-less number is dependant on the quantity of light energy absorbed and transmitted by the solution through the following equation:

A= -log T or A = 2-log %T

where T= transmittance= I/I⁰ where I is the absorbed light and I⁰ is the impinging light source.

Purpose

The purpose of this laboratory experiment is to practice, learn and carefully prepare molar solutions to investigate the relationship of concentration and spectrophotometer response. A 0.150 M solution of cobalt (II) chloride will be provided. You will prepare diluted solutions through the serial dilution method, measure the transmittance and calculate the absorbance values. You will prepare a Beerâs law graph of data; molar concentration vs. absorbance. Then determine Beerâs law constant using Microsoft Excel program.

You will also be given a concentrated sample of cobalt (II) chloride of unknown concentration and you must determine the appropriate dilution to prepare and thereafter determine the unknown concentration.

Procedure

In order to carry out this analysis procedure, it will be necessary to determine the wavelength of transition. Recall, that one specific energy maximum exists for each transition. We will study the transition in the range of 400- 600 nanometers (nm). This is the visible region of the spectrum.

Step 1. Prepare the following solution concentrations using the 0.150 M cobalt (II) chloride stock solution, 18 mm x 150 mm test tubes and Mohr pipets.

TABLE 1

Solution No.	0.150 M CoCl₂ (mL)	Volume water (mL)
1	5.0	0.0
2	4.0	1.0
3	3.5	1.5
4	3.0	2.0
5	2.5	2.5
6	2.0	3.0
7	1.0	4.0

Step 2. Carefully place a cork or rubber stopper over each solution and mix the contents.

Step 3. Measure the transmittance of a 0.150M solution between 400 nm and 600 nm at 25 nm intervals.

To accomplish this, follow these steps:

Caution: instruments are delicate. If you are uncertain about instrument operation, ask your instructor for assistance. (Instructions on instrument operation- see next page.)

SPECTRONIC is a registered trademark of Spectronic Instruments, Inc.

Instrument operation- edited version of product ^*Operatorâs Manual

Transmittance and Absorbance

1. Turn on the instrument by turning the Power Switch (10) clockwise. Allow the spectro-photometer to warm up for at least 15 minutes to stabilize.

2. After the warm-up period, set the desired wavelength with the Wavelength Control Knob.

3. Set the filter lever to the appropriate position for the selected wavelength (not required for SPECTRONICÂ® 20D).

4. Adjust the display to 0%T with the Zero Control (10). Make sure that the sample compartment is empty and the cover is closed.

5. Set the display mode to TRANSMITTANCE by pressing the MODE control key until the appropriate LED is lit.

6. Fill a clean cell with water and wipe the cell with a tissue (Kimwipe^TM) to remove liquid droplets, dust and fingerprints.

7. Place the cell in the sample compartment and align the guide mark on the cell with the guide mark at the front of the sample compartment. Press the cell firmly into the sample compartment and close the lid.

8. Carefully adjust the display to 100%T with the Transmittance/Absorbance Control (9). Move the knob slowly as you approach 100%T.

9. Remove the cell from the sample compartment and empty the water.

10. Rinse the cell twice with small volumes of the solution to be measured and fill it with the solution.

11. Wipe the cell with a tissue and insert the cell into the sample compartment. Align the guide marks and close the lid.

12. Read the appropriate value (%T) from the display.

13. Remove the cell from the sample compartment and repeat steps 10 through 12 for any remaining sample solutions.

14. When all measurements are completed, turn off the spectrophotometer by turning the Power Switchcounterclockwise until it clicks.

Step 4. Using solution no. 1, follow the directions for instrument operation and record (%) transmittance values for each wavelength in Table 2.

Remember to reset the instrument to 100% for each wavelength investigated.

DO NOT DISCARD THIS SOLUTION,

Step 5. The wavelength that gives the largest absorbance value will provide the greatest sensitivity to concentration change. Generate an Excel graph of wavelength (x) vs. absorbance (y) using these values. At zero absorbance, there isnât any CoCl₂ so donât forget to include the (0,0) data point. Connect your data points with a smooth curve. This is the absorption spectrum of aqueous CoCl₂. Submit this with your lab report.

Step 6. Reset your spectrometer using this wavelength (step 5), and measure this solution and the other six solutions listed in Table 1, and record your % transmittance values in Table 3.

TABLE 2

Wavelength (nm)	% Transmittance	Absorbance
400	88.3	0.055
425	73.7	0.132
450	43.6	0.362
475	23.8	0.623
500	15.7	0.804
525	16.0	0.797
550	38.9	0.410
575	73.0	0.37
600	85.3	0.069

MAXIMUM ABSORBANCE IS LOCATED AT __________500______nm.

TABLE 3 -Results measured at _500_____nm.

Test tube No.	% Transmittance	Absorbance	Molarity(M)
1	15.8	0.802
2	22.8	0.643
3	27.4	0.562
4	33.1	0.480
5	40.2	0.396
6	47.6	0.323
7	69.3	0.159

Step 7. Plot a graph of Molarity (M) vs. Absorbance for all seven solutions using Excel plotting techniques. This is called a calibration curve, whereby one establishes a measured response to a known solution concentration. Determine the slope of the line and include the linear equation of fit, and the correlation coefficient.

Recall the relationship A = kC or y = mx + b; where b is the (0,0) data point

Then: k = A/C

Step 8. The unknown solution to be analyzed is more concentrated than any of the standard solution concentrations that were measured to prepare the calibration curve.

Points to consider:

You must achieve a dilution such that the absorbance is within the absorbance range of the standards that were previously measured.

By using serial dilutions, will multiply many errors; thus it is best to obtain a dilution is one-step if possible.

Consider reducing the concentration by Â¾, or by Â½, and by Â¼ or to 0.10 of the original concentration and test the solution to see if the absorbance response is within range. You may also consider using the dilutions in Table 1. Once this is determined, proceed to the next step. Complete the dilution table for the diluted concentrations you prepared:

Trial	mL of unknown	mL of water	(%T)	A
1			51.9	0.284
2			70.7	0.151
3			79.3	0.101

Step 9. Using the desired dilutions, determined in step 8, measure two of the diluted solutions and record the measured transmittances in Table 4.

UNKNOWN NUMBER _____B_____

TABLE 4

Diluted concentration (M)	Transmittance (%T)	Absorbance (A)
Pure unknown B	8.7	1.062
1 mL of unknown in 1 mL of water	29.5	0.531

Calculation of absorbance, for diluted Unknown solution:

Step 10. Determine the concentration of the diluted solution using the linear equation of fit, knowing the absorbance and k, the slope of the line.

C = A/ k

Step 11: Determine the concentration in your original Unknown solution. Show your

back-calculation below to support your final answer to this experiment.

Calculations:

If needed I did a graph slope and got y= 0.1896x

R^2=0.9951

Unknown Solution No. ________ Concentration: ___________M

Prelab Questions

1. Consider a solution of 0.400M Co(NO₃)₂ provided by your lab instructor.

You are required to make the following dilute concentrations:

0.160M and 0.240M solutions. You are provided the following glassware: 5.0 mL Mohr pipet and 18 mm x 150 mm test tubes. Describe how you would prepare each solution, and support your description with your calculations. (Recall (M_c x V_c = M_d x V_d) where M_c is the molarity of the concentrated solution, V_c is the volume of the concentrated solution, M_d is the molarity of the dilute solution, and Vd is the volume of the diluted solution.)

Given 0.400 M Co(NO3)2 solution.

Lets say we pipette out 5 mL of this solution and dilute it to get 0.160 M solution. Then

Using, Mc x Vc = Md x Vd

0.400 x 5 = 0.160 x Vd or Vd= (0.400 x 5)/0.160 = 12.5 mL

Thus we need to add 7.5mL distilled water to 5 mL of cobalt nitrate. Since we are only provided with a 5 mL pipette, we can only measure out volumes in multiple of 5.

thus pipette out (5 x 2) 10 mL of cobalt nitrate solution in the test tube and dilute it by addition of (7.5 x2) 15 mL of distilled water.

Again for preparing 0.240 M solution

Lets say we pipette out 5 mL of this solution and dilute it to get 0.160 M solution. Then

Using, Mc x Vc = Md x Vd

0.400 x 5 = 0.240 x Vd or Vd= (0.400 x 5)/0.240 = 8.33 mL

Thus we need to add 3.33mL distilled water to 5 mL of cobalt nitrate. Again taking volumes only in multiples of 5, pipette out (5 x 3) 15 mL of cobalt nitrate solution in the test tube and dilute it by addition of (3.33 x3) 10 mL of distilled water.

2. In your own words, explain, in detail, what you will be learning during this lab. Address the chemical principles and, laboratory skills, in your answer.

Post - Lab Questions:

a) Letâs say, for a given concentration of salt solution, the maximum response at a wavelength maximum of 485nm, is 0.750. If an unknown solution is analyzed at a longer or shorter wavelength would the investigator obtain the same concentration for the Unknown solution? Explain why.

b) List all possible sources of errors that may result in this experiment.

CHEM 111L Lecture Notes - Lecture 6: Scatter Plot, Food Coloring, Molar Attenuation Coefficient

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