Professional Documents
Culture Documents
OrganicChemistrywithVernier
VernierSoftware&Technology
HO
HN
CH3
H3C
CH3
N
O
O
CH3
CH3
OH
O
H3C
OH
H3C
CH3
Acetylsalicylic acid
(aspirin)
Acetaminophen
Caffeine
Ibuprofen
OBJECTIVES
In this experiment, you will
MATERIALS
Part I Thin-Layer Chromatography
10 mL graduated cylinder
UV lamp (shortwave)
ethyl acetate
hexane
spotting capillary tubes
five 10 75 mm test tubes
test tube rack
unknown sample
mortar and pestle
filter paper
OrganicChemistrywithVernier
acetaminophen
acetylsalicylic acid
caffeine
ibuprofen
unknown sample
PROCEDURE
Part I Thin-Layer Chromatography
1. Obtain and wear goggles. Protect your arms and hands by wearing a long-sleeve lab coat and
gloves. Conduct this reaction in a fume hood
2. Obtain your unknown analgesic sample and dissolve approximately 50 mg in 2 mL of ethyl
acetate. Filter the solution through a pipet containing a cotton plug into a test tube.
3. Transfer approximately 12 mL of each pure compound to test tubes. Be sure to label each
test tube. Solutions of the pure compounds will be available in an ethyl acetate solution.
4. Prepare three development chambers using the 400 mL beakers and watch glasses.
a. Label the chambers 13. These will contain ethyl acetate, hexane, and
1:1 ethyl acetate/hexane, respectively.
b. Place a piece of filter paper against the side of the beaker. The filter paper will help
saturate the beaker with solvent vapors.
c. Fill each beaker with 510 mL of the developing solvent and cover with the watch glass.
5. Prepare the TLC plates.
a. Obtain three TLC plates and number the plates 13. Handle them carefully and by the
edges so that the adsorbent does not flake off.
b. Using a pencil (NOT an ink pen), lightly draw a line across the plate, approximately 1 cm
from the bottom. Across this line, evenly mark five places indicating the location where the
sample will be spotted, making sure they are not too close to the edge of the plate (see
Figure 1). Do this for all three TLC plates.
c.
Under each mark, lightly label each spot starting left to right as Ac, As, C, I, and
U. Repeat this step on the other TLC plates.
d.
Take a spotting tube and dip one end into the ethyl acetate solution containing
acetaminophen. Capillary action will draw the liquid into the tube.
e.
Lightly tap the tube on the mark for acetaminophen on all three TLC plates. Only a
small amount of sample needs to be delivered. The spot should be 12 mm in diameter.
f.
Repeat Steps de for the aspirin, caffeine, ibuprofen, and unknown sample on all
three TLC plates.
6. Place each of the TLC plate in the chamber and cover with the watch glass. The solvent level
must not be above the spots on the plate or your samples will dissolve into the solvent.
OrganicChemistrywithVernier
7. When the solvent has risen to within 1 cm from the top of the plate, remove the plate from the
chamber and with a pencil, gently draw a line to mark the position of the final solvent front.
8. After the plate has dried, observe the TLC plate under a UV lamp. Lightly outline the spots
with the pencil. CAUTION: Do not allow skin or eyes to come in contact with UV light.
Wear gloves, a lab coat, and UV resistant eye protection.
9. Identify the solvent mixture that gave the best separation and use that TLC plate to calculate
the Rf value for each spot. Record the values in your data table.
OrganicChemistrywithVernier
10. Obtain a small amount of acetaminophen. The solid should be in a powdered form. If it is
not, use a mortar and pestle to carefully grind the solid to a powder.
11. Prepare a sample for melting:
a. Pack a capillary tube 34 mm (~1/8 inch) deep with your sample by inserting the open end
into a small pile of the solid. A small amount of the solid will be pushed up into the tube.
b. Tap the closed end of the capillary tube on the table top to compress the sample into the
closed end.
c. Check the control knob on the Melt Station to confirm that it is in the Off position.
d. Carefully insert the capillary tube of solid into one of the sample holders of the Melt
Station.
12. Connect the Melt Station power supply to a powered electrical outlet.
13. Connect the Melt Station sensor cable to LabQuest or to a computer interface.
14. Start the data-collection program, then choose New from the File menu. You are now set
up to take melting temperature data for up to 20 minutes.
15. Use the prepared sample in a capillary tube to determine the melting temperature of the
sample.
a. Start data collection.
b. On the Melt Station, turn the control knob to the Rapid Heat setting.
c. Carefully observe the temperature vs. time graph. When the temperature is within
approximately 10C of the lowest possible melting temperature of your sample, turn the
control knob to a temperature setting corresponding to the expected melting temperature
provided in the table below.
Analgesic
acetaminophen
168172
134136
caffeine
ibuprofen
234236.5
7778
d. Carefully observe your sample. When the solid begins to melt, click Mark to mark the
temperature on your graph (or press the D key on the computer or the OK button on
LabQuest). When the entire solid has completely melted, click Mark again. The two values
marked on your graph describe the estimated melting temperature range of your substance.
When you are finished with this step, stop data collection.
e. Store the run by tapping the File Cabinet icon in LabQuest, or choosing Store Latest Run
from the Experiment menu in Logger Pro.
f. Discard the capillary tube and sample as directed by your instructor.
g. On the Melt Station, turn the control knob to the Fan/Cooling setting to get ready for the
next trial.
OrganicChemistrywithVernier
16. Prepare a new sample as described in Steps 1011 and repeat Step 15 for samples of
aspirin, caffeine, and ibuprofen.
Part III Test the Melting Temperature of Unknown Sample
17. Obtain a small amount of your unknown analgesic compound. The solid should be in a
powdered form. If it is not, use a mortar and pestle to carefully grind the solid to a powder.
18. Prepare a sample for melting:
a. Pack a capillary tube 34 mm (~1/8 inch) deep with your sample by inserting the open end
into a small pile of the solid. A small amount of the solid will be pushed up into the tube.
b. Tap the closed end of the capillary tube on the table top to compress the sample.
c. Carefully place the capillary tube of solid into one of the sample holders of the Melt
Station.
19. In the first trial, you will want to observe the melting process and make a rough estimate
of the melting temperature of your unknown sample. Dont worry if the heating rate is a bit
too rapid, and the sample melts too quickly. To do this:
a. Start data collection.
b. On the Melt Station, turn the control knob to a setting of 180C. The red light will turn on
indicating active heating.
c. Carefully observe your sample. If the solid begins to melt, click Mark to mark the
temperature on your graph. When the entire solid has completely melted, click Mark again.
The two values marked on your graph describe the estimated melting temperature range of
your substance.
d. If the solid does not melt by the time the temperature gets to 150C, turn the control knob
to the 220C setting. Continue observing your sample, and if the sample begins to melt,
mark the temperatures on the graph as previously described.
e. If the sample has not melted by the time the temperature gets to 190C, turn the knob to
the Rapid Heat setting. When the sample finally begins to melt, mark the graph as
previously indicated.
f. When you have determined the approximate melting temperature range for the sample,
stop data collection. Store the run by tapping the File Cabinet icon in LabQuest, or
choosing Store Latest Run from the Experiment menu in Logger Pro. Discard the capillary
tube and sample as directed by your instructor.
g. On the Melt Station, turn the control knob to the Fan/Cooling setting to get ready for the
next trial. The blue light will turn on indicating that the fan is cooling the Melt Station.
20. Now that you have a rough idea of the melting temperature, a more accurate
determination of the melting temperature can be made. Use a previously prepared sample in a
capillary tube, as described in Steps 1718, to determine the melting temperature of the
sample:
a. Start data collection.
b. On the Melt Station, turn the control knob to the Rapid Heat setting.
c. Carefully observe the temperature vs. time graph. When the temperature is within
approximately 10C of the lowest possible melting temperature of your sample, turn the
control knob to a temperature setting corresponding to your expected melting temperature.
d. Carefully observe your sample. When the solid begins to melt, click Mark to mark the
temperature on your graph. When the entire solid has completely melted, click Mark again.
OrganicChemistrywithVernier
DATA TABLE
Part I Thin-Layer Chromatography
Solvent mixture that gave
the best separation
Calculated Rf
acetaminophen
acetylsalicylic acid
caffeine
ibuprofen
unknown
Part II Melting Temperature of Analgesic Standards
Measured melting
temperature range
(C)
acetaminophen
acetylsalicylic acid
caffeine
ibuprofen
Part III Melting Temperature of Unknown Sample
Measured melting
temperature range
(C)
unknown analgesic
OrganicChemistrywithVernier
DATA ANALYSIS
1. Draw your TLC plate. Label each analgesic and show the calculation of the Rf value.
2. Based on your melting temperature data and the Rf value of the unknown, identify your
unknown analgesic.
OrganicChemistrywithVernier
INSTRUCTOR INFORMATION
1. Use a low intensity, shortwave UV lamp to examine the TLC plates.
2. Prepare the acetaminophen standard solution by dissolving 0.20 g per 25 mL of ethyl acetate.
3
Prepare the acetylsalicylic acid (aspirin) standard solution by dissolving 0.25 g per 25 mL of
ethyl acetate.
4. Prepare the caffeine standard solution by dissolving 0.25 g per 25 mL of ethyl acetate.
5. Prepare the ibuprofen standard solution by dissolving 0.25 g per 25 mL of ethyl acetate.
6. Crush tablets of Tylenol, Excedrin, Bayer, and Advil using a mortar and pestle. Choose
one as an unknown and distribute to each student or group.
7. The medicine tablets will not completely dissolve in ethyl acetate.
8. Prepare the 1:1 ethyl acetate/hexane solution by adding an equal volume of ethyl acetate and
hexane.
9. Some tailing may be seen on the TLC plate for the acetylsalicylic acid (aspirin) sample.
10. Small changes in the development solvent mixture can result in varying Rf values. Do not
let the developing chambers remain out for a long period of time.
11. Use TLC plates with a fluorescent indicator at a 254 nm excitation wavelength.
12. For best results, keep unused TLC plates in a desiccator.
13. Dispose of waste appropriately.
HAZARD ALERTS
Ethyl Acetate: Serious fire hazard (flash point 3.0C). Irritating to body tissues. Severe eye
irritant. Avoid all body tissue contact. Slightly toxic by ingestion. Vapor causes weakness, fatigue,
nausea and headache. Skin contact causes dermatitis. HMIS Classification: Health hazard2,
Flammability3, Physical hazard1.
Hexane: Serious fire hazard (flash point 23C). Severe eye and skin irritant. Moderately toxic by
ingestion. Vapor causes weakness, fatigue, nausea and headache. HMIS Classification: Health
hazard2, Flammability3, Physical hazard0.
Acetaminophen: May be harmful if inhaled and cause respiratory tract irritation. Moderately toxic
by ingestion. Skin and eye irritant. HMIS Classification: Health hazard2, Flammability0,
Physical hazard0.
Acetylsalicylic acid (aspirin): May cause respiratory tract irritation. Skin and eye irritant.
Moderately toxic by ingestion. HMIS Classification: Health hazard2, Flammability1, Physical
hazard0.
OrganicChemistrywithVernier
Caffeine: May cause respiratory tract irritation. Skin and eye irritant. Moderately toxic by
ingestion. HMIS Classification: Health hazard2, Flammability0, Physical hazard0.
Ibuprofen: May cause respiratory tract, eye and skin irritation. Moderately toxic by ingestion.
HMIS Classification: Health hazard1, Flammability0, Physical hazard0.
The hazard information reference is Sigma-Aldrich Co.,
1-800-325-3010, www.sigmaaldrich.com/safety-center/msds-search.html.
UV light: Protect your arms and hands by wearing a long-sleeve lab coat and gloves. Wear
appropriate eye protection. Contacts and normal eyeglasses do not provide protection. Acute
(short-term) effects include redness or ulceration of the skin. At high levels of exposure, these
burns can be serious.
COMPOUND INFORMATION
Compound
acetaminophen
acetylsalicylic acid
caffeine
ibuprofen
Compound
ethyl acetate
hexane (mixture of
isomers)
10
Chemical formula
Melting temperature
range (C)
C8H9NO2
168172
151.16
C9H8O4
134136
180.16
C8H10N4O2
234236.5
194.19
C13H18O2
7778
203.28
Chemical formula
Molar mass
(g/mol)
Density
(g/mL) at 25C
C4H8O2
88.11
0.902
C6H14
86.18
0.672
OrganicChemistrywithVernier
SAMPLE RESULTS
OrganicChemistrywithVernier
11
12
OrganicChemistrywithVernier