Aspirin Lab

Zhi Yang Lin

Lab Partners: Angelina Risi and Kwame Owusu

November 3, 2014
TA: Stephanie Barros
Chem 053-125

Purpose
The purpose of this two week aspirin lab is to synthesize aspirin, answer questions such as
percent yield and purity of the collected aspirin, provide an introduction to absorption
spectroscopy as an analysis tool, and introduce thin layer chromatography (TLC).
Procedure
Part 1: Aspirin Synthesis
2.001 g of salicylic acid and 5.0 mL of acetic anhydride was added to an erlenmeyer flask along
with approximately 5 drops of 85% phosphoric acid. The mixture was swirled around in the
flask, and any solid particles that were stuck to the sides of the flask were washed down with a
wash bottle containing distilled water. Shortly after, the solution was placed on a hotplate set to
75C. The solution was swirled occasionally while heating and after 10 minutes had elapsed, 2.0
mL of distilled water was added to the flask. The solution is heated for another 5 minutes; after a
total of about 15 minutes of heating on the hotplate and the reaction was thought to have come to
completion (signaled by the ceasing of vapors being produced), 20.0 mL of water was added and
the flask was allowed to cool to room temperature. Meanwhile, a Buchner funnel was set up. The
now room temperature flask was placed in an ice bath for around 5 minutes. The contents of the
flask were then filtered by using a Buchner funnel. The crystals leftover in the funnel were
washed thrice with cold distilled water with filtering after each washing period.
Part 2: Recrystallization
4.735 g of crude aspirin was obtained. Using a ratio of 6 g of crude aspirin to 20 mL of EtOH,
15.78 mL of EtOH was measured out and added to the crude aspirin in a flask. The solution was
then heated at low on a hotplate until all crystals have been dissolved. Meanwhile, on another
hotplate 50 mL of distilled water at 50C was being prepared. The 50 mL of warm water was
added to the solution; crystals formed and did not cease to form even under 75C heating. When
the mixture cooled to room temperature, it was placed in the ice bath once again to aid
recrystallization. The mixture was filtered again and washed with cold distilled water. The first
percent yield of the not completely dried aspirin sample was calculated to be 81.11%. A week
later after the aspirin sample was completely dried, a new percent yield of 36.6% was obtained.
Part 3: Purity Analysis of Aspirin
A spectrometer kit was calibrated and set up from an earlier spectroscopy skill building exercise.
In preparing aspirin samples for testing, exactly 1.000 grams of aspirin was measured into a 50
mL beaker with 5 mL of 95% ethanol. The mixture was quickly swirled and dissolved and 30 mL
of distilled water was added to the beaker and mixed again. The contents of the beaker were
transferred to a 50 mL volumetric flask. The beaker was thoroughly rinsed out with distilled
water and poured into the flask to ensure all contents of the beaker had been successfully
transferred. Additional distilled water was added to the volumetric flask until the solution level
reached 50 mL. The contents of the flask were mixed thoroughly then poured into a larger sized
beaker than the one. The flask was cleaned and properly dried before exactly 3 mL of the aspirin
solution was added to it along with enough 0.025 M of Fe(NO3)3 solution to obtain once again a
volume of 50 mL in the volumetric flask; the contents were mixed thoroughly again. The
absorbance level of this solution was measured to be greater than 1.5. A new aspirin solution was
needed. This time only 0.031 grams of aspirin was used in creating the aspirin solution and

again, 3 mL of this solution was used to create the mixture used to measure absorbance. The
second absorbance obtained was 1.338. A dilution of 1 mL of the mixture with 1.3381 absorbance
and 4 mL of M of Fe(NO3)3 solution was undertaken to yield a final absorbance of 0.298.
Spectroscopy: Skill Building Exercise
0.073 g of Salicylic Acid was placed in a beaker with 4.93 mL of ethanol to dissolve. After
dissolving, the mixture was transferred to a 100 mL volumetric flask and diluted with distilled
water to the 100 mL mark. The concentration obtained was 5.3*10-3 M. 5 mL of this solution was
then transferred into a 50 mL volumetric flask and diluted to the desired 50 mL mark with
Fe(NO3)3. The new concentration of this stock solution was obtained to be 5.3*10-4 M. Five clean
test tubes were properly cleaned, dried, and labeled with the contents that were going to be
poured into each. Test tube 1 contained 5 mL of stock solution and every test tube after that
contained (5-(n-1)) mL of stock solution and (n-1) mL of Fe(NO3)3, where n equals the number
of the test tube, until we reach test tube 5 which had 1 mL of stock solution and 4 mL of
Fe(NO3)3. The contents of each test tube were thoroughly mixed with a stirring rod that was
washed and dried between each mixing. Concentrations were solved for and recorded on a table.
Meanwhile, a spectrometer kit was set up, connected to a laptop, and calibrated with a cuvette
filled of the way with Fe(NO3)3. Once calibration was completed via Logger Pro, the contents
of the cuvette was dumped out, rinsed twice with one of the mixture from one of the test tube,
and filled of the way with the same mixture. The cuvette was then placed in the
spectrophotometer and a graph was run on Logger Pro. The wavelength at the peak of the graph
was recorded on the table with concentration. This was done for all five test tubes until all the
wavelengths were obtained and recorded. To find the best line equation, absorbance was graphed
versus concentration on Excel as a scatter plot and then a trend line was added. The line equation
obtained was y=1577.4x +0.0358.
Thin Layer Chromatography: Skill Building Exercise
0.5 mL of Ethanol was poured into two test tubes; 0.007 g of aspirin was mixed into one test tube
while 0.006 g of salicylic acid was placed into the other test tube and mixed. On a TLC plate, a
pencil line was drawn approximately 1cm from the bottom of the plate and marked with A and S
on each side, corresponding to aspirin and salicylic acid respectively. The aspirin and ethanol
mixture was poured onto a clean watch glass and with the stubby end of a toothpick the aspirin
and ethanol solution was dotted on the TLC plate. The TLC plate was dotted a couple more times
to make it more concentrated. The same was done for the salicylic acid but with a clean
toothpick. Both dots were allowed to dry and then placed into 150 mL beaker with about 5 mL of
mobile phase solvent, 80:20 hexanes: ethyl acetate mixture. The solvent line was confirmed to be
above the mobile phase before a watch glass was placed over the beaker. After the solvent had
run to about 1cm under the top of the TLC plate, the TLC plate was removed, a pencil line was
drawn to mark the place the solvent had reached, and heated on a hot plate previously set to 50C
for about a minute. Then the TLC plate was dipped into a potassium permanganate; then, the
back of the plate was dried slightly with a paper towel and heated again on the hot plate. The
distances the aspirin, salicylic acid, and the solvent travelled were all measured with a ruler and
the Rf values were calculated.

1 After calculations and further examinations, we determined that we had mixed up the aspirin
solutions and had used the mixture with an absorbance higher than 1.5 for dilution.

Data and Results

Figure 1: Spectroscopy Absorbance from TLC SBE

Note: The second to last curve is that of the diluted aspirin sample (1mL of aspirin solution with
4 mL of Fe(NO3)3) from above.
Figure 2: Concentration vs Absorbance from TLC SBE

Concentration vs Absorbance
1
0.8

f(x) = 1577.36x + 0.04

R = 1

absorbance
Linear (absorbance)

0.6

Linear (absorbance)

0.4
0.2
0
0

Table 1: Aspirin Synthesis and Recrystallization

Quantitative
amount
Salicylic Acid
2.001 g
Acetic Anhydride
5.0 mL

Weight Dish
Crude Aspirin and Weight
Dish
Crude Aspirin

2.867 g
7.602 g

Ethanol Needed to Dissolve

Crude Aspirin

15.78 mL

Theoretical Yield of Aspirin

Calculated
Weight Dish and Wet
Recrystallized Aspirin
Recrystallized Aspirin (wet)

2.610 g

Weight Dish and Dry

Recrystallized Aspirin
Dry Recrystallized Aspirin
Percent Yield of Wet
Recrystallized Aspirin
New Percent Yield of Dried
Recrystallized Aspirin

3.822 g

4.735 g

Observations (if any)

It took time for the salicylic acid to
dissolve in the acetic anhydride. After all
the heating was done and 20 mL of
distilled water was added, the solution
turned cloudy white, and when placed in
ice bath, the precipitate inside turns snow
like. C7H6O3(S)+C4H6O3(l)C9H8O4(S)
+C2H4O2(l)

After filtration, some crude aspirin was

filtered out along with the liquids. The
liquids were foam like originally but once
they were allowed to settle they appear to
be clear with an oily substance floating
above it. The crude aspirin left on top of
the funnel looks like mushy snow.
The crude aspirin dissolves completely in
within 3 minutes of coming in contact with
the ethanol. When 50 mL of warm water
came was added to the mixture afterwards,
crystals formed immediately and would not
stop even at 75C heating (not pure).
Contents were foamy when placed in the
ice bath again, and a second filtration
resulted in even more lost of aspirin.

4.984 g
2.117 g

0.955 g
81.11%
36.6%

Looks extremely watery and soft, almost

like Elmers glue
Very flaky and light

Table 2: Purity of Aspirin

Trial 1
Trial 2
Amount of Aspirin sample
0.100 g
0.031 g
Amount of 95% ethanol
5 mL
5 mL
Total amount of distilled water added to
Enough to make 50
Enough to make 50
volumetric flask with aspirin and 95% ethanol mL of the mixture
mL of the mixture
Amount of aspirin, distilled water, and ethanol 3 mL
3 mL
mixture transferred to a 50 mL cleaned
volumetric flask with (50-x) mL of Fe(NO3)3,
where x equals mL of aspirin, distilled water,
and ethanol mixture. (Let this solution be
called the aspirin solution)
Absorbance of aspirin solution
Greater than 1.5
1.338
Table 3: Dilution of Trial 1 Aspirin Solution for solving Purity of Aspirin Analysis
Quantitative Amount
Concentration of aspirin in a mixture of 5 mL of 95% ethanol and
0.011 M
distilled water totaling to a volume of 50 mL
Concentration of aspirin in aspirin solution
6.6*10-4M
Concentration of aspirin in a dilution with 1 mL of aspirin solution from 1.32*10-4M
the row above with 4 mL of Fe(NO3)3
New absorbance of diluted solution
0.298

Standard curve of absorbance (y) vs concentration of Salicylic Acid (x)

y= 1577.4x+0.0358
Concentration of Salicylic Acid in sample of diluted aspirin solution
1.66*10-4M
(1:4 of aspirin solution and Fe(NO3)3) with absorbance of 0.298
Concentration of Salicylic Acid in aspirin solution
8.3*10-4 M
Concentration of Salicylic Acid in aspirin sample, distilled water, and
0.0138 M
Iron (III) Nitrate mixture
Moles of Salicylic Acid in aspirin sample, distilled water and Iron (III)
6.9*10-4 moles
Nitrate mixture
Grams of Salicylic Acid in 50 mL of aspirin sample, distilled water and
0.095 grams
Iron (III) Nitrate
Percent purity of Aspirin sample
5%

: Originally, we thought we used a dilution of trail 2 to solve for purity, but in reality, the aspirin
solution for trial 1 was diluted.

:Standard curve is obtained later in the lab from the Spectroscopy skill building exercises but is
used in calculating purity of aspirin. This equation is illustrated in Figure 2 above.
Table 4: Spectroscopy SBE
Content
Stock solution (0.073 g of Salicylic Acid,
4.93 mL of Ethanol, and ~5mL of Fe(NO3)3
4 mL stock and 1 mL Fe(NO3)3
3 mL stock and 2 mL Fe(NO3)3
2 mL stock and 3 mL Fe(NO3)3

Concentration of Salicylic Acid (M)

5.3*10-4

Absorbance
0.867

4.2*10-4
3.2*10-4
2.1*10-4

0.690
0.518
0.367

1.06*10-4

1 mL stock and 4 mL Fe(NO3)3

Figure 3: TLC Plate

0.207

Note: Yellow represents aspirin,

white represents Salicylic acid, and
Z is the solvent.
Table 5: TLC SBE
Distance Traveled
1.3 cm
1.3 cm
3.6 cm

Aspirin
Salicylic Acid
Mobile Phase

Rf
0.36
0.36
N/A

Calculations
Note: Only one type of each calculation is shown.
Table 1: Sample Calculations
Mass of crude aspirin
crude aspirin+ weight dish=7.602 g

x+ 2.876 g=7.602 g

x=4.735 g

Amount of Ethanol Needed to Dissolve Crude Aspirin

6 g of Aspirin
4.735 g of Aspirin
4.735 20
=
; x=
=15.738 mL of EtOH
20 m L of EtOH
6
x ( mL )

Theoretical Yield of Aspirin

C4H6O3(l) + C7H6O3(s) C9H8O4(s) + C2H4O2(l)
2.001 g C 7 H 6 O3

1mol C 7 H 6 O3
1mol C 9 H 8 O4 180.17 g C 9 H 8 O4

=2.610 g C 9 H 8 O4
138.13 g C7 H 6 O3 1 mol C7 H 6 O3
1 mol C 9 H 8 O4

Calculating Percent Yield

Percent yield=

Actual yield
0.955 g
100 =
100 =36.6
theoretical yield
2.610 g

Table 3: Sample Calculations

Calculating Molarity given a mass and volume (Note. Moles and volume can also be calculated
using the molarity formula, given the value of the other two variables)
0.100 g Aspirin

1 mol Asprin
=5.55 104 mol
180.157 g Aspirin

n 5.55 104 mol

M= =
=0.011 M
V
0.05 L

Calculating new concentration given an original volume and concentration and a new volume
C1 V 1=C 2 V 2

0.011 M ( 3 mL )=C 2 ( 50 mL )

C2 =6.6 104 M

Calculating concentration of Salicylic Acid given absorbance value

Note: y is absorbance and x is concentration
y=1577.4 x +0.0358

0.298=1577.4 x +0.0358

x=

0.2980.0358
=1.66 104 M
1577.4

Calculating grams given moles

6.9 104 mol of Salicylic Acid

138.121 g of Salicylic Acid

=0.095 g o f Salicylic Acid
1mol of Salicylic Acid

Calculating percent purity

purity=

mass of pure product 0.100 g Aspirin sample0.095 g Salicylic Acid

;
=5
mass of impure product
0.100 g Aspirin sample

All calculations from Table 4 were obtained via Logger Pro

Table 5 Sample Calculations
Calculating Retention Factor
Rf =

distance spot traveled

1.3 cm
=
=0.36
distance solvent traveled 3.6 cm

Discussion

For the percent yield of our recrystallized aspirin, we first obtained 81.11%. However,
our second percent yield, the real percent yield, was only 36.6%. This dramatic drop in percent
yield is due to the fact that the aspirin sample was not given enough time to dry completely after
the second filtration. The first percent yield calculated the mass of the aspirin sample along with
the mass of water molecules thus leading to a larger yield. The reason our actual percent yield
was so low can be attributed to many factors. The most apparent one is the lost of crude aspirin
in the first and second filtration processes. This in turn can be attributed to not letting the aspirin
sample sit long enough in the ice bath to form firm enough crystals and washing the aspirin
samples with distilled water that is not cold enough to prevent the dissolving of the aspirin
sample during filtrations, causing some aspirin particles to be filtered away.
In addition, our sample was expected not to be very pure at all. When the crude aspirin
was dissolved in ethanol and warm distilled water was added and heated at constant 75C,
crystals still continued to form. We concluded, with the help of our TA, that this sample was
indeed not very pure, so a 5% purity was not really surprising at all. The TLC plate for the
aspirin sample and salicylic acid further confirms this point. The retention factor was similar for
both dots, indicating a massive amount of salicylic acid present in our aspirin sample. If this had
not been the case, the aspirin dot shouldve traveled further upwards since it is less polar than the
salicylic acid. In addition, when we added Iron (III) Nitrate to the ethanol and aspirin sample
mixture, the solution turned dark purple and the absorbance rate was way greater than 1.5; this
again reiterates that there is a massive amount of salicylic acid in our sample. We were suppose
to use the aspirin sample from the second trail to calculate purity, but the samples were mixed up
and the difference could not be explained until later when percent purity was calculated using the
amount of aspirin in the second trail, 0.031 g, which resulted in negative purity percentages
every time. After many failed attempts of calculating purity, we realized that we had disputed
over which aspirin sample contained 0.031 grams during lab and upon further review, had chosen
incorrectly. The error in purity couldve been caused by many factors. First, some of the
glassware and lab equipment used werent properly washed between usages, namely dipping an
unwashed stirring rod into one of our solutions causing contamination. Second, due to constant
mistakes, time was constrained and we began to rush and even more mistakes resulted from
rushing and trying to catch up. Third, it is possible that in the very beginning, we did not put in
enough acid catalyst or waited long enough to allow the reaction to go to completion. Additional
heating time, slightly more acid catalyst, staying cool in the heat of the moment, and properly
cleaned and dried glassware couldve lead to a higher purity percent.
Real world techniques that were observed were mainly using spectroscopy to identify
amount of a particular substance in an aqueous product, using a Buchner funnel to filtrate and
extract solids precipitates in a solution, and recrystallizing products to yield higher purities.
Spectroscopy can be used in real life in forensics to determine x amount of a chemical in say a
victims blood. Spectroscopy enables us to calculate the concentration of a particular substance
in a clear non-murky solution, which in turns allows us to calculate moles Buchner funnel is self
explanatory in that larger versions of these funnels can filter out a valued precipitate from an

unwanted solvent. Recrystallizaing is largely used in medicine purifications.2 It ensures that no

other harmful excess reagents are present in the product sample. Such reagents can cause a
multiple of harm to the human body and could damage the body more than the medicine actually
intend to heal.
Post Lab Questions
1. Our TLC and spectroscopy tests coincide. The absorbance was extremely high and the
TLC plate practically showed no difference between retention factors of the aspirin and
salicylic acid. There was an extremely large amount of salicylic acid present in the aspirin
sample (0.095 g out of 0.100 g were salicylic acid). Aside from error, we may also have
cross contaminated the aspirin solution. We accidentally dipped an unclean stirring rod
that was used to stir another solution in our aspirin sample. Other causes may result from
a lower amount of acid catalyst that was called for and shorter heating time on the hot
plate in the beginning steps of the procedure; both of these factors together can lead to the
reaction not going to completion.
2. Compound A, compound B, compound C, compound D.
3. a. If the product was washed with hot water during filtrations, some of the aspirin
A
sample would dissolve in the water and will be filtrated down along with the liquid.
This will in turn result in a lower percent yield.
C
b. If the aspirin is left out to dry for weeks, it will react to the moisture in the air and
yield salicylic acid, which smells like vinegar, as stated in the lab lecture. This process
is called hydrolysis. If this is allowed to happen, the aspirin sample would have a
lower purity percentage since there will be more salicylic acid present than before.
H2O(l)+C9H8O4(s)C2H4O2(l)+C7H6O3(s)
4. a. Salicylic acid was chosen as the limiting reagent for several reasons. It is extremely
more expensive than acetic anhydride making it cost effective, it is a solid making it
easier to measure out for experiments, and it is less dangerous making it easier to handle
in experiments.
b. C7H6O3(S)+C4H6O3(l)C9H8O4(S)+C2H4O2(l)
Acetic anhydride has a density of 1.08g/mL and a molar mass of 102.09g/mol, thus 0.5
mL would equal 0.54 grams of acetic anhydride or .005 mol. Since the mole ratio
between salicylic acid and acetic anyhdride is 1:1, as long as ~2.00 grams of salicylic
acid is equal to 0.005 mol or less, salicylic acid should still be the limit reagent and the
theoretical yield should not change. However, 2.00 grams of salicylic acid is greater than
0.005 mol (0.014 mol) thus making the acetic anhydride the limiting reagent. This means
only 0.005 mol of salicylic acid will react leading to a smaller theoretical yield when
compared to all 0.014 mol of salicylic acid reacts.
5.

8 H atoms

22 H atoms

8 H atoms

2 http://www.intechopen.com/books/recrystallization/recrystallization-of-drugs-significance-on-pharmaceuticalprocessing

B
D

6. Diacetyl is made naturally through fermentation3. It is the product of yeast converting

glucose to ethanol in the fermentation process4. One can identify how much diacetyl is
present in a mixture of ethanol and glucose by using spectroscopy. When using
spectroscopy, the blank cuvette should be just a mixture of glucose and ethanol to allow
for the best wavelength absorbance to show up in LoggerPro. Creating dilutions from a
stock solution will ultimately give us a standard curve in the form of Beers Law. From
there, concentrations can be calculated. Given a particular volume from before, number
of moles can be calculated and converted to grams.

3 http://en.wikipedia.org/wiki/Diacetyl

4 http://www.toxipedia.org/display/toxipedia/Diacetyl