Lab 4_TLC Report
docx
School
Pennsylvania State University *
*We aren’t endorsed by this school
Course
203
Subject
Chemistry
Date
Apr 3, 2024
Type
docx
Pages
5
Uploaded by CountComputer15136
Thin Layer Chromatography of Analgesics – Final Report
Report (70 points)
Commercial tablet used:
You: unknown #1
Partner: unknown #2
1.
Draw a representative TLC plate showing each standard compound (caffeine, acetaminophen, ibuprofen, and aspirin (acetylsalicylic acid)). Calculate the R
f value for each compound. (Show calculations). (10 pts)
All of the above Rf values are reported under 60 ethyl acetate: 40 hexane mixed solvent ratio conditions. Rf = distance traveled by the compound / distance traveled by the solvent.
2.
Also draw a representative plate of your
unknown tablet. Calculate the R
f value for each component. (Show calculations). (5 pts)
All of the above Rf values are reported under 60 ethyl acetate: 40 hexane mixed solvent ratio conditions.
3.
What compound(s) were in each of the two
tablets in your group? (5 pts)
unknown #1 has acetaminophen (after conducting an experimental plate that limits the drug selection down to acetaminophen/ibuprofen/and unknown 1; upon seeing the similarities between acetaminophen and unknown 1 in Rf, conducting another co-spotting test, to see whether acetaminophen would map on to the spot of unknown 1 exactly will give us a more definitive answer)
unknown #2 has ibuprofen (both share similar Rf values when compared across trials, similar Rf means both compounds have similar polarity in the given solvent, suggesting that these two may have been similar compounds)
4.
Describe what happened with the different solvent combinations: 60:40 ethyl acetate:hexane, hexane, and ethyl acetate.
Explain qualitatively why
you got these results with the different solvents. (What properties of the solvents led to this behavior?) (15 pts)
If 60:40 ethyl acetate:hexane mixed solvent served as the baseline (where ibuprofen had an Rf value of 0.80). Then in ethyl acetate solvent alone, compounds like aspirin, ibuprofen, and caffeine traveled the furthest (for instance, ibuprofen had an Rf of 0.87). On the other hand, these compounds traveled the smallest distance in pure hexane (i.e. ibuprofen only had an Rf value of 0.17). We got these results due to the intermolecular forces between the compounds and the solvents. The 60:40 mixture provides a balance between polar and nonpolar characteristics as it combines ethyl acetate (polar) with hexane (nonpolar). The mixture's moderate polarity allows compounds like ibuprofen, aspirin, and caffeine to travel
intermediate distances, creating the optimal separation between spots. Since ethyl acetate is a polar solvent, it has a strong affinity (dipole-dipole) for polar compounds like ibuprofen, aspirin, and caffeine, enabling them to travel easily along the plate. However, this ease of movement may lead to poor separation and thus it would be hard to differentiate between different compounds by their polarity if many spots overlap. Hexane, on the other spectrum,
is too nonpolar and leads to little to no movement of the compound samples because it fails to break the dipole-dipole interactions between the relatively polar compound of the silica gel. Thus, it would also be cumbersome to differentiate between the slightest separation. As such, the optimal solvent to use in this experiment is 60:40 ethyl acetate:hexane mixture to allow for optimal separation, visualization, and collection of the Rf values.
5.
A TLC was run in 70:30 ethyl acetate:hexane for compounds C and D. Compound C has an R
f
of 0.5 and compound D has an R
f
of 0.3 in this solvent system. (10 pts)
a.)
Draw the TLC plate. Which compound is more polar? Why?
A higher Rf value represents that the compound is less polar because of its increased interactions with the mobile phase (ethyl acetate and hexane) than the silica gel (stationary phase). Thus, the compound that is the more polar will interact with the stationary phase more and thus move a shorter distance, which, in this case, is compound D because (0.3 < 0.5). b.)
You decide to run another TLC on compounds C and D in 30:70 ethyl acetate:hexanes on a silica gel plate. Would the R
f
values for the compounds increase or decrease? Why?
The Rf values for the compounds would decrease. Since ethyl acetate is more polar than hexane, by increasing the ratio of hexane to ethyl acetate, you are decreasing the polarity of the solvent. With more nonpolar hexanes, it’s less likely to interfere with the polar silica gel (stationary phase); thus, making it unlikely to displace any of the compounds that are bonded to
the silica gel. As such, by increasing the level of hexanes in the solvent, the compounds will travel significantly less, lowering the Rf value, because they are interacting more with the stationary phase (silica gel) than the mobile phase (30:70 mix solvent), especially for the polar compounds that have a higher affinity for other polar molecules. 6.
In a TLC experiment, why should the spot not be immersed in the solvent in the developing chamber? (5 points)
The spot should not be immersed in the solvent in the beginning because there is a higher chance of
the compound being dissolved into the solution at the bottom of the beaker rather than traveling upwards in the plate (controlled by the IMFs between the compound and the mobile and stationary phase). This creates contamination and produces inaccurate results.
7.
Explain why the diameter of the initial spot should be as small as possible. (5 points)
The diameter of the initial spot should be as small as possible because, with a smaller dot, it’s more clear (with a more defined shape) when you see spot separation as the smaller dots
only have to travel a little to see the distinction. If the spots were larger, their movement upwards may interfere with one another and potentially cause mixing or streaking due to the
high concentration of compounds. With a smaller spot, you will also have higher accuracy and precision in determining the center of the spot and thus produce a more accurate Rf value than if the spots were large.
8.
Why is it important to not let the solvent run off the top of the plate? (5 points)
If you let the solvent run off from the top of the plate, then you can’t measure the total distance the solvent has traveled since there is no final measurable position (solvent line).
Your preview ends here
Eager to read complete document? Join bartleby learn and gain access to the full version
- Access to all documents
- Unlimited textbook solutions
- 24/7 expert homework help
Since Rf is the distance traveled by the compound/distance traveled by the solvent. As such,
we cannot find the accurate Rf value if we let the solvent run off the top (lacking the accurate measurement of the solvent travel distance).
9.
A student obtained the TLC results below. What are the R
f
values for Compound X and Compound Y? Are they the same compound? How could the student verify this (what techniques could be used?) (10 points)
Rf for compound X is around 1.35/2.80 cm = 0.48
Rf for compound Y is around 1.40/2.80 cm = 0.50
Because Rf for X is slightly lower than the Rf of Y, we cannot conclude that they are the same compound. The student could verify the identity of the compounds by conducting co-spotting or alternating spots. By conducting co-spotting, the student would test three lanes (only X, only Y, and X and Y) with emphasis on the last lane. If there is only one distinct dot, then X and Y are the same compound. But they are not the same if there is an elongation or blobby spot. Alternatively, the student could experiment with alternating spots where using a wide plate of four or more lanes, he can alternate between X and Y and observe the trend in line. If the line is straight across, then the two compounds are the same. If there is a zig-zag or wavy line across, then the two compounds are different.
X
Y
Related Documents
Related Questions
Table of chemicals and reagents with their physicochemical properties (name,
molecular formula, molecular weight, pKa, melting point, boiling point, etc.)
Write a short outline or a flow diagram of the procedure
List and safety issues
Read and understand the MSDS information for each chemical used in this practical
Experimental procedure
Please do not scratch the clear sides of the UV cuvettes as this will invalidate your results.
Part 1: Preparation of a calibration curve
A calibration curve should be constructed using at least five concentrations of the pure paracetamol standard
supplied in the range 3-15 ug/mL as follows:
Weigh about 150 mg of paracetamol accurately and transfer to a 200 mL volumetric flask. Add 50 mL of o.1 M
NaOH and dilute to about 100 mL with deionised water (DI). Shake and make up to volume with DI water. Dilute
10 mL of the resulting solution to 100 mL (volumetric flask) with DI water (= stock solution). Prepare standard
solutions containing 3, 6, 9, 12 and 15…
arrow_forward
According to the following TLC, do you see any evidence on the TLC that product was lost?
a
Yes, TLC shows the product is also in the mother liquor (filtrate).
b
Yes, TLC shows product in the crude sample.
c
No, TLC is not quantitative.
d
No, there is no evidence on the TLC that the product was lost
arrow_forward
Three students did a chromatography experiment, where Rf = distance of solute / distance of solvent.
What could be the possible errors why student 3 had results that are quite far from that of students A and B?
arrow_forward
Use the simulation lab to obtain the distance for the solvent front and the distance for each spot and then
calculate the Rf
35ig figs
acetaminoph
Unknown
Unknown
Sample
caffeine
ibuprofen
aspirin
en
Mixture #1
Mixture #2
13.0 cm13.0 cm 13. O cm 13.0 cm
13.0cm
Distance to
130 cm
solvent front
1.77cm 1.89cm
11.1
12.6 cm
1.78cm la.6 cm 11.0 cm
6.27cm
Distance
Spot(s) traveled
5.58cm
Calculated R:(s)
Components of
Unknowns
D. Additional Exercises
1. Which of the substances tested is most polar? What parts of the structure of this substance is polar?
2. Which of the substance tested is most non-polar? What parts of the structure of this substance in non-polar?
arrow_forward
Column Chromatography
Alumina
Chromatography Mixture
9:1 Hexanes:Ether
8:2 Hexanes:Ether
1:1 Hexanes:Acetone
Amount Used
3.962 g
0.143 g
9.50 mL
9.50 mL
11.00 mL
Additional Observations (Color, etc.)
BIU X₂ X² →
BI IU X₂ X² →
BI IU X₂ X² →
BI IU X₂ X² →
BIU X₂ X² →
arrow_forward
Given the following chromatogra
What molecules in the mixture were separated successfully in Chromatogram A? In Chromatogram B?
From the given data, what are the differences between the two chromatograms?
arrow_forward
Color BEFORE adding
Color AFTER adding
Intensity of color AFTER
adding FeCl3
Test Tube #
FeCl3
FeCl3
#1 (salicylic acid)
clear
lilac
100%
#2 (commercial aspirin A) clear
slightly pink
50%
#3 (commercial aspirin B) clear
lilac
95%
#4 (aspirin from your
clear
pink
20%
synthesis above)
#5 (control)
clear
clear
0%
Based on this data and the melting point of Aspirin, how pure is the synthesized product?
arrow_forward
Please correct answer and don't use hand rating
arrow_forward
how to get uncertainty?
arrow_forward
Table #2 Worktable for BCA Assay
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
appropriate standard. Pipette each of the protein samples directly into the water, do not
dribble on the side of the test tubes. The standard is at a concentration of 2 mg/ml and
contains bovine serum albumin (BSA). Calculate the amount of BSA protein added to each
of the tubes and write this in Table 2 (Protein (ug)).
arrow_forward
Question: Using a sample of aspirin for HPLC analysis. If the aspirin was contaminated (wet) with trace amounts of a solvent such as ethyl acetate, hexane, or water, would this contaminant be detected and appear as a peak in the HPLC chromatogram? Briefly explain based on the type of detector used in HPLC analysis and what type of compounds can be detected.
My Answer: The HPLC analysis detects chromophores, compounds with multiple double bonds. Since water, ethyl acetate, and hexane all lack multiple pi bonds this will not be detected in the HPLC.
-Question for you, is this complete (and correct), and should I include anything about UV detection? Is UV detection used in HPLC? I thought it was only used on TLC to show compunds that arent visable.
arrow_forward
Question 3 (b)
Given the chromatogram shown below, answer the following questions:
(b) Calculate the adjusted retention time for compound/peak 3 (in minutes).
3.
10
0.
5
8.
10
Time (min)
HO
OH
Benzoic acid
Phenol
Toluene
Benzene
Detector Signal (mAU)
20
arrow_forward
Answer number 8
arrow_forward
10:36 1
ull ?
Search
Expert Q&A
Done
Sample G.C. :
Simple Distillation
Fraction 3
1500-
1000-
500-
Time (min)
AM Simple- F3
Sample GC of products obtained from Simple
distillation of solution that is 40% ethyl acetate and
60% propyl acetate.
calculate the following for each sample gas
chromatograph.
a) retention time for each peak (label peaks)
b) area of each peak c) %composition
arrow_forward
1. The concentration of NaOH from the best three of your trials.
2. The average concentration of 3 of your trials (only use 3 trials)
3. The standard deviation of your 3 trials [If you have a calculator with stats
on it, you can use that/bring it to class, but for the lab report you will have
to write out your calculations.]
4. Your PPT.
arrow_forward
QUESTION 5
In a chromatographic separation using a polar stationary phase and a
mobile phase composed of 60% hexane and 40% ethyl acetate, two
compounds, benzene (peak A) and toluene (peak B), are eluted. The width
of peak A at its base is 1.2 min, the width of peak B at its base is 1.3 min,
and the retention time of peak A is 5.2 min and peak B is 3.5 min.
What is the resolution (Rs) between these two peaks and is it acceptable?
R = 1.36. It is not an acceptable resolution.
R = 0.74. It is an acceptable resolution.
R = 1.36. It is an acceptable resolution.
R = 0.74. It is not an acceptable resolution.
arrow_forward
In the Thermo Fisher application note about wine analysis (Lesson 3), the following
chromatogram was collected of nine components of wine. If peak 3 has a retention time of
3.15 minutes and a peak width of 0.070 minutes, and peak 4 has a retention time of 3.24
minutes and a peak width of 0.075 minutes, what is the resolution factor between the two
peaks? [Hint: it will help to review Lesson 2 for this question.]
MAU
300
200
T
34
5
100-
1 2
CO
6
7
8
9
0
2.4
2.6
2.8
3.0 3.2 3.4
3.6
3.8 4.0 4.2
4.4
4.6
4.8
5.0
5.2
Minutes
3.22
0.62
1.04
O 1.24
arrow_forward
2
how do you find the answer and what is the answer for each component using the volume of first table?
arrow_forward
In the Thermo Fisher application note about wine analysis (Lesson 3), the following
chromatogram was collected of nine components of wine. If peak 3 has a retention time of 3.15
minutes and a peak width of 0.070 minutes, and peak 4 has a retention time of 3.24 minutes and
a peak width of 0.075 minutes, what is the resolution factor between the two peaks? [Hint: it
will help to review Lesson 2 for this question.]
mAU
300
200
100
0
3.22
0.62
1.04
1.24
2.4 2.6
2.8
LO
34
12
ww
3.0
5
3.2 3.4 3.6
3.8 4.0
Minutes
7
8
9
4.2 4.4 4.6 4.8 5.0
5.2
arrow_forward
SEE MORE QUESTIONS
Recommended textbooks for you

Chemistry
Chemistry
ISBN:9781305957404
Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher:Cengage Learning

Chemistry
Chemistry
ISBN:9781259911156
Author:Raymond Chang Dr., Jason Overby Professor
Publisher:McGraw-Hill Education

Principles of Instrumental Analysis
Chemistry
ISBN:9781305577213
Author:Douglas A. Skoog, F. James Holler, Stanley R. Crouch
Publisher:Cengage Learning

Organic Chemistry
Chemistry
ISBN:9780078021558
Author:Janice Gorzynski Smith Dr.
Publisher:McGraw-Hill Education

Chemistry: Principles and Reactions
Chemistry
ISBN:9781305079373
Author:William L. Masterton, Cecile N. Hurley
Publisher:Cengage Learning

Elementary Principles of Chemical Processes, Bind...
Chemistry
ISBN:9781118431221
Author:Richard M. Felder, Ronald W. Rousseau, Lisa G. Bullard
Publisher:WILEY
Related Questions
- Table of chemicals and reagents with their physicochemical properties (name, molecular formula, molecular weight, pKa, melting point, boiling point, etc.) Write a short outline or a flow diagram of the procedure List and safety issues Read and understand the MSDS information for each chemical used in this practical Experimental procedure Please do not scratch the clear sides of the UV cuvettes as this will invalidate your results. Part 1: Preparation of a calibration curve A calibration curve should be constructed using at least five concentrations of the pure paracetamol standard supplied in the range 3-15 ug/mL as follows: Weigh about 150 mg of paracetamol accurately and transfer to a 200 mL volumetric flask. Add 50 mL of o.1 M NaOH and dilute to about 100 mL with deionised water (DI). Shake and make up to volume with DI water. Dilute 10 mL of the resulting solution to 100 mL (volumetric flask) with DI water (= stock solution). Prepare standard solutions containing 3, 6, 9, 12 and 15…arrow_forwardAccording to the following TLC, do you see any evidence on the TLC that product was lost? a Yes, TLC shows the product is also in the mother liquor (filtrate). b Yes, TLC shows product in the crude sample. c No, TLC is not quantitative. d No, there is no evidence on the TLC that the product was lostarrow_forwardThree students did a chromatography experiment, where Rf = distance of solute / distance of solvent. What could be the possible errors why student 3 had results that are quite far from that of students A and B?arrow_forward
- Use the simulation lab to obtain the distance for the solvent front and the distance for each spot and then calculate the Rf 35ig figs acetaminoph Unknown Unknown Sample caffeine ibuprofen aspirin en Mixture #1 Mixture #2 13.0 cm13.0 cm 13. O cm 13.0 cm 13.0cm Distance to 130 cm solvent front 1.77cm 1.89cm 11.1 12.6 cm 1.78cm la.6 cm 11.0 cm 6.27cm Distance Spot(s) traveled 5.58cm Calculated R:(s) Components of Unknowns D. Additional Exercises 1. Which of the substances tested is most polar? What parts of the structure of this substance is polar? 2. Which of the substance tested is most non-polar? What parts of the structure of this substance in non-polar?arrow_forwardColumn Chromatography Alumina Chromatography Mixture 9:1 Hexanes:Ether 8:2 Hexanes:Ether 1:1 Hexanes:Acetone Amount Used 3.962 g 0.143 g 9.50 mL 9.50 mL 11.00 mL Additional Observations (Color, etc.) BIU X₂ X² → BI IU X₂ X² → BI IU X₂ X² → BI IU X₂ X² → BIU X₂ X² →arrow_forwardGiven the following chromatogra What molecules in the mixture were separated successfully in Chromatogram A? In Chromatogram B? From the given data, what are the differences between the two chromatograms?arrow_forward
- Color BEFORE adding Color AFTER adding Intensity of color AFTER adding FeCl3 Test Tube # FeCl3 FeCl3 #1 (salicylic acid) clear lilac 100% #2 (commercial aspirin A) clear slightly pink 50% #3 (commercial aspirin B) clear lilac 95% #4 (aspirin from your clear pink 20% synthesis above) #5 (control) clear clear 0% Based on this data and the melting point of Aspirin, how pure is the synthesized product?arrow_forwardPlease correct answer and don't use hand ratingarrow_forwardhow to get uncertainty?arrow_forward
- Table #2 Worktable for BCA Assay 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 appropriate standard. Pipette each of the protein samples directly into the water, do not dribble on the side of the test tubes. The standard is at a concentration of 2 mg/ml and contains bovine serum albumin (BSA). Calculate the amount of BSA protein added to each of the tubes and write this in Table 2 (Protein (ug)).arrow_forwardQuestion: Using a sample of aspirin for HPLC analysis. If the aspirin was contaminated (wet) with trace amounts of a solvent such as ethyl acetate, hexane, or water, would this contaminant be detected and appear as a peak in the HPLC chromatogram? Briefly explain based on the type of detector used in HPLC analysis and what type of compounds can be detected. My Answer: The HPLC analysis detects chromophores, compounds with multiple double bonds. Since water, ethyl acetate, and hexane all lack multiple pi bonds this will not be detected in the HPLC. -Question for you, is this complete (and correct), and should I include anything about UV detection? Is UV detection used in HPLC? I thought it was only used on TLC to show compunds that arent visable.arrow_forwardQuestion 3 (b) Given the chromatogram shown below, answer the following questions: (b) Calculate the adjusted retention time for compound/peak 3 (in minutes). 3. 10 0. 5 8. 10 Time (min) HO OH Benzoic acid Phenol Toluene Benzene Detector Signal (mAU) 20arrow_forward
arrow_back_ios
SEE MORE QUESTIONS
arrow_forward_ios
Recommended textbooks for you
- ChemistryChemistryISBN:9781305957404Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCostePublisher:Cengage LearningChemistryChemistryISBN:9781259911156Author:Raymond Chang Dr., Jason Overby ProfessorPublisher:McGraw-Hill EducationPrinciples of Instrumental AnalysisChemistryISBN:9781305577213Author:Douglas A. Skoog, F. James Holler, Stanley R. CrouchPublisher:Cengage Learning
- Organic ChemistryChemistryISBN:9780078021558Author:Janice Gorzynski Smith Dr.Publisher:McGraw-Hill EducationChemistry: Principles and ReactionsChemistryISBN:9781305079373Author:William L. Masterton, Cecile N. HurleyPublisher:Cengage LearningElementary Principles of Chemical Processes, Bind...ChemistryISBN:9781118431221Author:Richard M. Felder, Ronald W. Rousseau, Lisa G. BullardPublisher:WILEY

Chemistry
Chemistry
ISBN:9781305957404
Author:Steven S. Zumdahl, Susan A. Zumdahl, Donald J. DeCoste
Publisher:Cengage Learning

Chemistry
Chemistry
ISBN:9781259911156
Author:Raymond Chang Dr., Jason Overby Professor
Publisher:McGraw-Hill Education

Principles of Instrumental Analysis
Chemistry
ISBN:9781305577213
Author:Douglas A. Skoog, F. James Holler, Stanley R. Crouch
Publisher:Cengage Learning

Organic Chemistry
Chemistry
ISBN:9780078021558
Author:Janice Gorzynski Smith Dr.
Publisher:McGraw-Hill Education

Chemistry: Principles and Reactions
Chemistry
ISBN:9781305079373
Author:William L. Masterton, Cecile N. Hurley
Publisher:Cengage Learning

Elementary Principles of Chemical Processes, Bind...
Chemistry
ISBN:9781118431221
Author:Richard M. Felder, Ronald W. Rousseau, Lisa G. Bullard
Publisher:WILEY