TLC, IR, MP TEMPLATE
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NEW TEMPLATE: TLC, FTIR, AND MELTING POINT Pre-lab Questions Procedure and Safety 1.
(10 pts) Write a VERY brief bullet point list summary of the procedure. The point of this is to make sure you read and understand the procedure. You do not need to include every detail. •
TLC= use 3 jars (100 hexanes, 85% hexanes/15%ethyl acetate 10%ethyl acetate) Prepare and spot the plate, add to jar, wait, visualize using UV light. Do same with our compounds but with the 85%/15% mix. •
FTIR=Run computer program •
MELTING POINTS= Find melting point, use the machine like in intro lab 2.
(4 pts) To prepare 10 mL of a 80% hexanes/20 % ethyl acetate developing solvent, 8 mL of hexanes is measured in a 10 mL graduated cylinder and then 2 mL of ethyl acetate is added. Explain how to prepare 10 mL of a 85% hexanes/15% ethyl acetate developing solvent. 8.5ml of hexane and 1.5ml of ethyl acetate. 3.
(4 pts) Write one sentence summarizing the safety concerns for this experiment. Everything used in this experiment is toxic or hazardous in some way. 4.
(4 pts) What do you need to be careful of when picking up a TLC spotter? Where should spotters be put when you are finished? In the glass receptacle. Part 1: Choosing the best developing solvent for TLC 5.
(15 pts total, 5 pts each structure, 5 pts picking most polar) Draw the bond-line structures of the two solids you separated, 9-fluorenone and benzoic acid. Which is more polar? Benzoic acid is more polar 6.
(5 pts) Given that the surface of a silica coated TLC plate (the type we use) can be represented by the structure below, would 9-fluorenone or benzoic acid be more attracted to the TLC plate surface? Flourenone 7.
(15 pts total, 5 pts ea structure, 5 pts correct labeling) In our Ochem labs, two solvents are often used for developing TLC plates: ethyl acetate and hexanes. Draw the bond-line structures for these two solvents. (Just draw one isomer of hexane). Which of these solvents is polar and which is nonpolar? Polar= ethyl acetate 8.
(10 pts) As you learned in the tutorial video, “TLC Theory”, how fast a compound moves up a plate is determined by its solubility in the mobile phase (developing solvent) vs its attraction to the
stationary phase (silica). If a nonpolar solvent is used as developing solvent, would you predict 9-
fluorenone or benzoic acid moves up the TLC faster? Why? Benzoic acid is nonpolar and more attracted to the polar TLC plate. 9.
(4 pts) Consider the photo below showing TLC plates from a different experiment. Which plate shows the best separation for the four different compounds that were spotted? Part 2: Functional groups and interpreting FTIR spectra 10.
(40 pts, 4 pts each name, 4 pts each peak) Fill in the following table. Compound Structure Functional Group Name Location of key IR peaks (cm
-1
) alkenes 1680-1600 Hydroxyl/alcohols 3650-3200 Carboxyl/ ketones 1780-1650 Carboxyl/ carboxylic acids 3300-2500 Benzene ring/aromatics 1600-1430 11.
(8 pts) What are the functional groups in 9-fluorenone? Predict the two main peaks you will see above 1500 cm
-1
in the FTIR spectrum for 9-fluorenone. Plate A Plate B Plate C
Ketone and benzine ring/aromatics 12.
(12 pts) What are the functional groups in benzoic acid? Predict the three main peaks you will see above 1500 cm
-1
in the FTIR spectrum for benzoic acid. Carboxylic acids, benzne ring, ketones or alkenes Data and Observations Part 1: TLC 1.
(15 pts) Insert a photo of your three TLC plates used for testing developing solvents with provided solutions of benzoic acid and 9-fluorenone. The solvent front should be marked and circles drawn around any spots visualized using UV light. Plates should be clearly labeled with developing solvent used and what was spotted. Photo should include a ruler for calculating R
f
values. 2.
(15 pts) Insert a photo of the TLC plate showing results from your separated benzoic acid. The solvent front should be marked and circles drawn around any spots visualized using UV light. Plates should be clearly labeled with developing solvent used and what was spotted. Photo should include a ruler for calculating R
f
values. 3.
(15 pts) Insert a photo of the TLC plate showing results from your separated 9-fluorenone The solvent front should be marked and circles drawn around any spots visualized using UV light. Plates should be clearly labeled with developing solvent used and what was spotted. Photo should include a ruler for calculating R
f
values. Part 2: FTIR
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Color AFTER adding
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The retention time of the toluene sample was 14.50 min.
The standard deviation of the peak was 7.0542 (s).
2a. The mobile phase velocity was?
0.0806 mm/s
0.1344 mm/s
1.344 mm/s
1.344 x 10³ mm/s
(i)
(ii)
(iv)
2b. The number of theoretical plates was?
(i)
(ii)
(iii)
(iv)
2c. The height equivalent to a theoretical plate was?
1.041x10³ m
1.015x105 m
9.862x10-6 m
1.186x10¹² m
(i)
(ii)
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(iv)
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(ii)
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(ii)
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concentration(C)(mg/ml)
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elution time (time)
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1
1
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302.85
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5
1
7.53
15.83
benzoic acid
1
1
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89.98
saccharin
1
1
1.91
84.86
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1
4.47
69.58
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a.
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b. The equation for the trendline (y = mx + b) is displayed on the graph. Which variable, in the equation,
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A1
feMass
B
D
Mass
Volume
2.
1,8
2,19
Mass vs. volume
3
3,15
4,08
4,13
5,57
7.
6,7
8,34
6.
y= 0,794x-0.064
8
6.
10
11.
2
4.
10
13
14
15
Figure 1: Plotting of Mass vs. Volume of a Substance.
bttps://www.chemisalforums.com/index.php?toric=69575.0
Life University
CHM 111
March 2021
Default Page Style
English (USA)
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1 and 2 answers?
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20-200 microliter pipette
100-1000 microliter pipette.
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Standard
Water (μl)
Tube #
1
2
3
4
5
6
7
8
9
0
125
325
175
325
325
325
400
400
BSA source (μl)
300 stock
375 stock
325 stock
175 of tube #2
325 of tube #3
325 of tube #5
325 of tube #6
100 of tube #7
0
Protein (ug)
Add the volumes listed in Table 2 to each tube. Add the water first and then the
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Table 1: The result for test tube 1 to 8 using of BSA stock solution.
Tube
BSA Cone
H;O (ml)
BSA (ml)
Biuret
ABS
mg/ml
reagent (ml)
1.
1.0
4.
0.
2.
0.9
0.1
4.
0.003
3.
2.
0.8
0.2
4.
0.012
4.
3.
0.7
0.3
4.
0.021
5.
4.
0.6
0.4
4.
0.028
6.
0.5
0.5
4.
0.039
7.
6.
0.4
4.
0.042
8.
7.
0.3
0.7
4.
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Protein
Tube
Uknown (ml)
H;0 (ml)
Biuret reagent
ABS
concentration
(mg/ml)
(ml)
9.
4.
0.036
10.
4.
0.035
11.
0.5
0.5
4.
0.017
12.
0.5
0.5
4
0.016
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Multiple Choice
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EVEN
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