Professional Documents
Culture Documents
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RESULTS AND CALCULATIONS ...................................................................................................... 39
CALCULATION OF RF VALUES ........................................................................................................ 39
DISCUSSION ......................................................................................................................................... 40
CONCLUSION ....................................................................................................................................... 42
REFERENCES ....................................................................................................................................... 43
APPENDIX ............................................................................................................................................. 44
APPENDIX B ......................................................................................................................................... 48
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CHAPTER ONE
INTRODUCTION
Aspirin (acetylsalicylic acid) is a synthetic organic compound derived from salicylic acid.
Salicylic acid is a natural product found in the bark of the willow tree and was used by the ancient
Greeks and Native Americans, among others, to counter fever and pain. However, salicylic acid is
A German chemist named Felix Hoffman is credited with being the first to synthesize
aspirin in 1897. Hoffman's father had severe arthritis but could not tolerate salicylic acid he was
taking for pain relief. The name given for Hoffman's new compound was A-spirin. Apparently
this comes from acetylation (A-), together with Spirin, part of the name for Meadow-sweet
Friedrich Bayer, the employer of Hoffman, patented the name and began marketing the
product in 1899. It was a huge success and sales grew rapidly. Bayer's company set up by himself,
is generally reckoned to have been the first pharmaceutical company, and the production of aspirin
is generally accepted to have laid the foundation of the modern pharmaceutical industry.
In this experiment, aspirin (acetylsalicylic acid, C9H8O4 ) was synthesized, purified, and
the percentage yield determined. The purity of the product was confirmed by measuring its melting
point range.
3
The reaction that is used for the synthesis is shown below. In this reaction, an excess of
acetic anhydride (C4H6O3) was added to a measured mass of salicylic acid (C7H6O3) in the
presence of a catalyst, sulfuric acid (H2SO4). The mixture was heated to form the acetylsalicylic
acid (C9H8O4) and acetic acid (C2H4O2). After the reaction took place, water was added to destroy
the excess acetic anhydride and cause the product to crystallize. The aspirin was then collected,
4
Washing Bottle Melting Point Apparatus
OBJECTIVES:
1. 5g of dry salicylic acid and 7mL of acetic anhydride were placed in a 200mL Erlenmeyer
flask. Three (3) drops of concentrated sulphuric acid were added. The flask was swirled
2. The mixture was warmed in a water bath to about 50-60˚C with stirring for about 15
3. 75mL of distilled water was added to the mixture, stirred and filtered at the pump.
4. The product was recrystallized by dissolving the solid in about 15mL of hot alcohol and
5. The clear solution obtained was allowed to cool slowly, enabling the crystals to separate
6. The crystals were filtered at the vacuum pump, allowed to drain well and dried in an oven
at 50˚C.
5
7. The yield was determined as follows:
𝟐.𝟔𝟑𝟎𝟑𝐠
% Yield =𝟒.𝟓𝟎𝟖𝟏𝐠 × 100
= 58.3461%
1. 0.5M HCl was standardised using analar sodium carbonate (Na2CO3) and the standardized
2. Twenty (20) tablets of aspirin were weighed individually and collectively to determine the
mass of each tablet and the total mass of the twenty tablets in order to determine the weight
distribution. The tablets were then powdered using mortar and pestle.
4. 50mL of 0.5M NaOH was added to the flask, which was immersed in boiling water-bath
5. The solution was cooled and the excess back-titrated with 0.5M HCl phenol red as
indicator.
6. A blank determination was carried out by heating, cooling and titrating a further 30mL of
0.5M NaOH. The difference between the titrations represented the amount of alkali
required by the aspirin. Methyl orange was used as an indicator for the standardization of
the NaOH.
6
RESULTS AND CALCULATIONS
Standardisation of HCl
Reaction equation:
Burette Reading/mL 1 2 3
(𝟐𝟎.𝟎𝟎+𝟏𝟗.𝟖𝟎)𝐦𝐋
Average titre = = 19.90mL
𝟐
Milliequivalence Calculation
𝟏𝟎𝟎
This implies the mass of 100% Na2CO3 = × 2.65g
99
7
Determining the factor of HCl
𝐀𝐜𝐭𝐮𝐚𝐥 𝐰𝐞𝐢𝐠𝐡𝐭
F(Na2CO3) = 𝐍𝐨𝐦𝐢𝐧𝐚𝐥 𝐰𝐞𝐢𝐠𝐡𝐭
𝟐.𝟔𝟖𝟎𝟎𝐠
= 𝟐.𝟔𝟕𝟔𝟖𝐠
= 1.0012
𝐅(𝐍𝐚𝟐𝐂𝐎𝟑)× 𝐕(𝐍𝐚𝟐𝐂𝐎𝟑)
F(HCl) = 𝐕(𝐇𝐂𝐥)
𝟏.𝟎𝟎𝟏𝟐×𝟐𝟎𝐦𝐋
= = 1.0062
𝟏𝟗.𝟗𝟎𝐦𝐋
Burette Reading/mL 1 2 3
(𝟐𝟏.𝟔𝟎+𝟐𝟏.𝟔𝟎)𝐦𝐋
Average titre = = 21.60mL
𝟐
Milliequivalence Calculation
8
9.125g HCl in 500mL ≡ 0.5M NaOH
𝟏𝟎𝟎
This implies mass of 100% HCl = × 9.125g
𝟑𝟔
= 25.3472g HCl
𝐦𝐚𝐬𝐬
Density (𝛒) =𝐯𝐨𝐥𝐮𝐦𝐞
𝐦𝐚𝐬𝐬
Volume =𝐝𝐞𝐧𝐬𝐢𝐭𝐲(𝛒)
Density = 1.18g/mL
𝟐𝟓.𝟑𝟒𝟕𝟐𝐠
Volume =𝟏.𝟏𝟖𝐠/𝐦𝐋 = 21.4806mL
The 21.50mL HCl was diluted with distilled water to the 500mL mark of the 500mL volumetric
flask.
Preparation of NaOH
9
10g in 500mL ≡ 0.5M NaOH
𝟏𝟎𝟎
Mass of NaOH to be weighed = × 10g = 10.4167g
𝟗𝟔
The 10.4551g NaOH was dissolved in distilled water and topped up to the 500mL mark of the
volumetric flask.
F(HCl) = 1.0062
V(HCl) = 21.60mL
F(NaOH) = ?
V(NaOH) = 20mL
𝐅(𝐇𝐂𝐥)× 𝐕(𝐇𝐂𝐥)
F(NaOH) = 𝐕(𝐍𝐚𝐎𝐇)
(𝟏.𝟎𝟎𝟔𝟐)× (𝟐𝟎𝐦𝐋)
= (𝟐𝟏.𝟔𝟎𝐦𝐋)
= 0.9317
10
Table 3: Results from the Determination of Blank
Burette Reading/mL 1 2 3
𝟑𝟐.𝟏𝟎+𝟑𝟐.𝟐𝟎
Average titre = = 32.15mL
𝟐
Volume of HCl that neutralised 30mL of blank (NaOH without aspirin) = 32.15mL
i. 0.5052g
ii. 0.5047g
Volume of HCl that reacted with the synthesised aspirin = (32.15 – 20.20) mL
= 11.95mL
Equivalent volume of NaOH that reacted with the synthesised aspirin = 11.95mL × 1.0062
= 12.0241mL
11
𝟏𝟐.𝟎𝟐𝟒𝟏𝐦𝐋
Then 12.0074mL NaOH ≡ × 0.045g = 0.5411g
𝟏𝐦𝐋
𝐦𝐚𝐬𝐬 𝐨𝐟 𝐩𝐮𝐫𝐞
% Purity of synthesised aspirin =𝐦𝐚𝐬𝐬 𝐨𝐟 𝐢𝐦𝐩𝐮𝐫𝐞 × 100
𝟎.𝟓𝟒𝟏𝟏
= 𝟎.𝟓𝟎𝟓𝟐 × 100 = 107.11%
Volume of NaOH that reacted with the ASA = (32.15 – 20.10) mL×1.0062
=12.1247 mL = 12.12 mL
𝟏𝟐. 𝟏𝟐𝟒𝟕𝐦𝐋
∴ 12.1247 mL ≡ × 0.045g
𝟏 𝐦𝐋
= 0.5456g
𝟎.𝟓𝟒𝟓𝟔𝐠
% Purity of the synthesised aspirin = 𝟎.𝟓𝟎𝟓𝟐𝐠 × 100
= 107.99%
𝟏𝟎𝟕.𝟏𝟎+𝟏𝟎𝟕.𝟗𝟗
Average % purity of synthesised aspirin = = 107.55%
𝟐
12
PERCENTAGE CONTENT OF ASPIRIN TABLETS
𝐱−𝐭
Weight/g Deviation (x-t) Percent Deviation (| | × 𝟏𝟎𝟎)/%
𝐭
13
20 0.3861 -0.0016 0.4127
7.7544g⁄
Average weight per tablet = 20 = 0.3877g
If 0.3877g ≡ 0.3g
𝟎.𝟓𝐠
Then 𝟎.𝟑𝐠 × 0.3877g = 0.6462g
i. 0.6467g
ii. 0.6470g
iii. 0.6464g
1 2 3
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CALCULATION OF PERCENTAGE CONTENT OF TABLETS
Volume of HCl needed to neutralize 30mL of blank (NaOH without aspirin) = 32.15 mL
𝟎.𝟔𝟒𝟔𝟕𝐠
Therefore 0.6467g aspirin powder contains 𝟎.𝟔𝟒𝟔𝟐𝐠 × 0.50g = 0.5004g ASA
Equivalent volume of NaOH that reacted with aspirin = 12.05 mL× 1.0062 = 12.1247 mL
𝟏𝟐.𝟏𝟐𝟒𝟕𝐦𝐋
12.1247 NaOH ≡ × 0.045g = 0.5456g
𝟏𝐦𝐋
𝟎.𝟓𝟒𝟓𝟔𝐠
% Content of Aspirin Tablet = 𝟎.𝟓𝟎𝟎𝟒𝐠 × 100g = 109.033%
𝟎.𝟔𝟒𝟕𝟎𝐠
0.6470g ≡ × 0.5g = 0.5006g
𝟎.𝟔𝟒𝟔𝟐
15
= 11.9235mL
𝟏𝟏.𝟗𝟐𝟑𝟓𝐦𝐋
11.9235mL ≡ × 0.045g = 0.5366g
𝟏𝐦𝐋
𝟎.𝟓𝟑𝟔𝟔𝐠
% Content of Aspirin tablet = 𝟎.𝟓𝟎𝟎𝟔𝐠 × 100 = 107.19%
If 0.6462g ≡ 0.50g
𝟎.𝟔𝟒𝟔𝟒𝐠
Then 0.6464g ≡ 𝟎.𝟔𝟒𝟔𝟐𝐠 × 0.50g = 0.5002g
Equivalent weight of NaOH that reacted with aspirin = 12.35 mL × 1.0062 = 12.4266 mL
𝟏𝟐.𝟒𝟐𝟔𝟔𝐦𝐋
12.4266mL ≡ 𝟏𝐦𝐋
× 0.045g = 0.5592g
𝟎.𝟓𝟓𝟗𝟐𝐠
% Content =𝟎.𝟓𝟎𝟎𝟐𝐠 × 100 = 111.795%
𝟏𝟎𝟗.𝟎𝟑𝟑+𝟏𝟎𝟕.𝟏𝟗𝟎+𝟏𝟏𝟏.𝟕𝟗𝟓
Average percentage content = = 109.34%
𝟑
16
DISCUSSION OF RESULTS
Aspirin, a drug widely known for its analgesic, antipyretic, and anti-inflammatory
properties, is a compound derived from two acids namely acetic acid and salicylic acid.
The experiment consisted of three (3) parts: (1) the preparation of the pertinent solutions,
(2) the standardization of 0.50M NaOH and 0.50M HCl solution, and (3) the analysis of the aspirin
sample and the percentage content of aspirin tablets. The first part of the experiment was the
preparation of Acetylsalicylic Acid (Aspirin). A white, milky mixture was obtained when salicylic
acid, acetic anhydride and sulphuric acid (a catalyst) were mixed. The mechanism of the reaction
is:
This shows that the oxygen in salicylic acid attacks one of the carbons in acetic anhydride. Also,
the mechanism shows how acetic acid was separated from the acetylsalicylic acid.
17
In the first part of the experiment, heating of the mixture was done and a clear yellow liquid
was obtained. Heating was employed so that salicylic acid would melt and react with acetic
anhydride. On the other hand, water was added after heating (not at the start of the experiment).
This was to prevent the reaction of acetic anhydride with water at the start of the experiment, if
this had happened, no aspirin could have formed. In this manner, acetic anhydride was decomposed
After cooling to room temperature and adding distilled water, the liquid became whitish
(cloudy) with white precipitates. This addition of water is very important in purification and
isolation of the crystals from the liquid. Since aspirin is insoluble in cold water, only the acetic
anhydride was decomposed by the water. Purification was needed to eliminate any salicylic acid
and acetic anhydride that did not react, as well as the acetic acid product and sulphuric acid. In this
part, purification was not yet complete (it was continued on the recrystallization part). Isolation
was done through vacuum filtration and white, needle-like crystals were obtained.
The crude/impure product was then weighed and it weighed 4.5081g.There was a
continuation of the purification process. The recrystallization was done by dissolving the product
in 15mL of hot alcohol and poured into 40mL of water, which was allowed to cool slowly. The
cooling process enabled the aspirin to crystallise out as white needle-like crystals, leaving behind
the impurities.
acid, the hydrolysis of the sample by alkali into neutrality was required. The hydrolysis of the
aspirin often uses large amounts or excess NaOH because such a reaction is slow and sufficient
18
amount of NaOH reacting with acetylsalicylic acid would just yield water as the product. The
whole process is called back-titration. It is a process in which the excess of standard solution used
are often required when the rate of reaction between the analyte and reagent is slow or when the
The melting point range (135˚C - 136˚C) obtained indicated that the synthesised aspirin
was pure since it matched the specification in the British Pharmacopoeia. The percentage purity
calculated was 106.95%, meanwhile the BP range is 99.5% - 101.0%.This shows there could be
impurities in the aspirin synthesised. The deviation as compared to BP value could be due to faulty
titration and some amount of moisture that could not be eliminated after drying the product.
From the calculations, the average percentage content of the aspirin tablets was 109.34%.
This was quite a deviation from the specified range (95% – 105%) indicated by the British
These errors may include inaccurate weighings, probable faulty titration. The balance was not
really giving stable values. These constitute the sources of error from which the deviation could
have stemmed.
CONCLUSION
The synthesised aspirin was impure because its percentage purity of 106.9% exceeded the
BP specification. The aspirin tablet also failed. The percentage content of the tablets exceeded
19
CHAPTER TWO
SPECTROPHOTOMETRY
INTRODUCTION
Discovery of Paracetamol
molecule (acetanilide) was added to a patient's prescription about 100 years ago. But since
acetanilide is toxic in moderate doses, chemists modified its structure to try and find a compound
that was less harmful but which still retained the analgesic properties. One of these compounds is
(from para-acetyl-amino-phenol) in the UK. When mixed with codeine it goes by the trade name
Tylenol.
20
Acetanilide Paracetamol Aniline
In fact, in the body, the original compound, acetanilide is partially converted into a mixture
of paracetamol and aniline. The paracetamol provides the painkilling properties, but the aniline is
toxic. Paracetamol has a very similar structure to aspirin, and because of this they are recognised
by the same enzyme. This enzyme is responsible for the biosynthesis of prostaglandins, which are
involved in the dilation of blood vessels that causes the pain experienced in a headache. Reduction
of the amount of prostaglandin, therefore, helps prevent headaches and other pain.
Preparation of Paracetamol
Paracetamol is one of the most common drugs used in the world, and is manufactured in
huge quantities. The starting material for the commercial manufacture of paracetamol is phenol,
which is nitrated to give a mixture of the ortho and para-nitrotoluene. The o-isomer is removed
by steam distillation, and the p-nitro group reduced to a p-amino group. This is then acetylated to
give paracetamol.
21
Paracetamol as Poison
Because paracetamol is a potent drug that is available without prescription, it is often used
in suicide attempts, and in this respect it is potentially more dangerous than other over-the-counter
drugs such as aspirin. This is because paracetamol overdoses often cause liver failure, and there
have been many cases where attempted suicides have awakened from an overdose and changed
their minds, yet still died a few days later from liver damage.
The reasons for this poisoning are to do with the process by which paracetamol is
22
This compound is extremely toxic, and like other such compounds is eliminated in the liver
by reaction with a tripeptide, glutathione. If insufficient glutathione is available, the toxic quinone
will not be eliminated and begins to react with cellular proteins and nucleic acids in the liver,
However, two compounds, methionine and N-acetylcysteine can boost levels of the vital
glutathione in the liver, and so can be used as antidotes for paracetamol poisoning if the overdose
23
is discovered in time. A new formulation of paracetamol is now being marketed in the UK which
incorporates methianine, such that the drug carries its own antidote with it!
OBJECTIVES:
PROCEDURE
1. 0.1006g of paracetamol powder was weighed and used to prepare the following
concentrations:
0.00025%
0.0050%
0.00075%
0.001%
0.0015%
4. The slope of the curve was found and coefficient of correlation determined.
TABLETS
24
2. 0.15g of the paracetamol powder was added to 50mL of 0.1M NaOH, diluted with
100mL of water, and shaken for about five minutes. Sufficient water was then
3. The resulting solution was mixed properly, filtered and diluted with water to 100mL
4. 10mL of the resulting solution was added to 10mL of 0.1M NaOH and diluted to
CURVE
0.1g of paracetamol was dissolved in 100mL 0.1M NaOH to give 0.1% paracetamol
solution
0.01
× 100mL = 10mL
0.1
25
10mL of the 0.1% solution of paracetamol was diluted with0.1M NaOH to100mL to produce
0.001
× 100mL = 10mL
0.01
10mL of the 0.01% paracetamol was diluted to 100mL with 0.1M NaOH to give the
0.0015
× 100mL = 15mL
0.01
15mL of 0.01% paracetamol solution was diluted with 0.1M NaOH to yield the required
0.00075
× 100mL = 50mL
0.0015
50mL of the 0.0015% (prepared in ii above) was diluted with 0.1M NaOH to 100mL to
0.00025
× 100mL = 25mL
0.001
25mL of the 0.001% (prepared in i above) was diluted with 0.1M NaOH to 100mL to
v. To prepare 0.00050%,
0.0005
× 100mL = 5mL
0.01
26
5mL of already prepared 0.01% paracetamol solution was diluted with 0.1M NaOH to 100mL to
27
19 0.6629 0.0065 0.9902
𝟏𝟑.𝟏𝟐𝟖𝟒
Average weight of 20 tablets = = 𝟎. 𝟔𝟓𝟔𝟒𝐠
𝟐𝟎
RESULTS
CONCENTRATION/% ABSORBANCE
0.00025 0.176
0.00050 0.352
0.00075 0.469
0.0010 0.698
0.0015 0.967
28
CALIBRATION CURVE
1.2
1 y = 638.54x + 0.0216
R² = 0.9928
0.8
Absorbancee
0.6
0.4
0.2
0
0 0.0002 0.0004 0.0006 0.0008 0.001 0.0012 0.0014 0.0016
Concentration/%
29
The equation from the graph is
y =638.54x + 0.0216
Slope Determination
A = ƐbC………………….. 1
y = mx + c……………….. 2
c = 0.0216 (intercept)
C = concentration = x, Ɛ = absorptivity
This implies,
𝟎.𝟒𝟗𝟖−𝟎.𝟎𝟐𝟏𝟔
x= 𝟔𝟑𝟖.𝟓𝟒
x = 7.4608 × 10-4
30
This implies that the concentration of paracetamol in the tablet is 7.4608 × 10-4%.
𝟕.𝟒𝟔𝟎𝟖 × 𝟏𝟎−𝟒
Therefore the % content of the tablet = × 𝟏𝟎𝟎
𝟕.𝟓 × 𝟏𝟎−𝟒
= 99.4773%
DISCUSSION
concentration of analyte in a series of standard solutions should be a straight line with an intercept
Sufficiently low concentrations of the analyte and the standard were prepared for the following
reasons:
i. The Beer–Lambert law is valid only for low concentrations of the analyte. As such very
low concentrations were used in the analysis. This is because at higher concentrations,
the individual particles of each analyte no longer behave independently of each other.
The resulting interaction between the particles of the analyte may change the analyte’s
absorptivity.
ii. A second contribution is that the analyte’s absorptivity depends on the sample’s
refractive index. Because the refractive index varies with the analyte’s concentration,
the values of the absorptivity and the molar absorptivity may change. For sufficiently
31
low concentrations of the analyte, the refractive index is essentially constant and the
A blank was also used in measuring the absorbance to correct the transmittance for
the sample in order to account for the loss of radiation due to scattering, reflection or absorption
The percentage content obtained for the paracetamol tablets is 99.5%. The
paracetamol passed successfully according to the BP. The BP range is 95.0% to 105.0%.
CONCLUSION
The percentage content of the paracetamol tablets was approximately 99.5%, which fell in
the BP range. The UV Spectrophotometric method is therefore quite an efficient method for the
assay of paracetamol tablets. The solvent NaOH used was also very useful for the determination.
32
CHAPTER THREE
CHROMATOGRAPHY
INTRODUCTION
components in a sample by distribution of the components between two immiscible phases namely
a stationery phase and a mobile phase. The stationery phase consists of a fixed bed of small
particles with a large surface area while the mobile phase is a fluid that moves constantly through,
or over the surface of the stationery phase. The components are separated because of their
selectively retarding the passage of some compounds through the stationery phase while allowing
others to move more freely. Therefore by determining the retardation of each separated compound
an evaluation of the separation can be achieved. A factor namely the Retardation Factor (Rf) for
each of the separated or eluted substances has then been introduced to enable the qualitative
defined as a measure of that fraction of the total elution time that it spends in the mobile phase. In
terms of parameters easily obtainable from the chromatogram, the Rf is defined as the ratio of the
33
Thin-layer chromatography, one of the chromatographic techniques, involves the same
In this case, however, the solid adsorbent is spread as a thin layer (approximately 250 um) on a
plate of glass or rigid plastic. A drop of the solution to be separated is placed near one edge of the
plate, and the plate is placed in a container, called a developing chamber, with enough of the eluting
solvent to come to a level just below the "spot." The solvent migrates up the plate, carrying with
it the components of the mixture at different rates. The result may then be a series of spots on the
plate, falling on a line perpendicular to the solvent level in the container (see, for example, the
following Figure). The retention factor (Rf) of a component can then be measured as indicated in
34
This chromatographic technique is very easy and rapid to perform. It lends itself well to
the analysis of mixture composition and may also be used to advantage in determining the best
eluting solvent for subsequent column chromatography. However, volatile compounds (bp <
The same solid adsorbents used for column chromatography may be employed for TLC;
silica gel and alumina are the most widely used. The adsorbent is usually mixed with a small
amount of "binder"--for example, plaster of Paris, calcium sulfate, or starch--to ensure adherence
of the adsorbent to the plate. The plates may be prepared before use, or commercially available
pre-layered plastic sheets may be used. In either case, the plates or sheets are dried for an hour or
more in an oven at 110EC before use. This procedure removes moisture that is adsorbed on the
adsorbent and produces a surface that is more effective in adsorbing and separating the components
of the mixture being analyzed. The relative eluting abilities of the various solvents that may be
used are the same. The eluting power required of the solvent is directly related to the strength of
35
A distinct advantage of TLC is the very small quantity of sample required. A lower limit
of detection of 10-9 g is possible in some cases. The spot of sample is applied to the plate with care,
which is normally not large, for the same reason that the amount of sample placed at the top of a
column should not be too large. However, larger samples such as 0.5 mg may be used on larger
TLC plates, which have thicker coats of adsorbent. In these cases isolation of the components
may be achieved by scraping the separated spots from the plate and eluting (extracting) them with
an appropriate solvent. It is usually necessary to repeat this process a number of times in order to
obtain even several milligrams of material. Such a procedure would offer a method of
identification of the various components, however, because enough material could be collected to
Often it is useful for the purpose of identification to run TLC chromatograms of knowns
and unknowns side by side. Other useful applications involve running multiple aliquots of samples
collected from a chromatographic column or samples taken from a reaction mixture in order to
Detection of spots on the chromatogram is easy for colored materials, and a number of
procedures are available for locating spots of colorless materials. For example, irradiation of the
plate with ultraviolet light will permit location of the spots of compounds that fluoresce.
Alternatively, the solid adsorbent may be impregnated with an otherwise inert, fluorescent
substance. Spots of materials that absorb ultraviolet light but do not fluoresce will show up as
black spots against the fluorescing background when the plate is irradiated with ultraviolet light.
Other detecting agents are more often used. These agents may be sprayed onto the chromatograms,
causing the spots to become readily apparent. Examples of detecting agents used in this way are
36
sulphuric acid, which causes many organic compounds to char, and potassium permanganate
solution. Iodine is another popular detecting agent. In this case the plate is placed in a vessel whose
atmosphere is saturated with iodine vapor. Iodine is adsorbed by many organic compounds, and
their spots on the chromatogram become colored (usually brown). Since these spots usually fade,
it is a good idea to circle the spots with a pencil while they are still visible in order to have a
Under a given set of conditions (adsorbent, solvent, layer thickness, and homogeneity) the
rate of movement of a compound with respect to the rate of movement of the solvent front, Rf, is
a property of that compound. The value is determined by measuring the distance traveled by a
substance from a starting line in the before-mentioned figure. The property has the same
Chromatank
50ml Measuring Cylinder
UV Lamp
Three 10cm × 20cm Aluminum TLC
Filter Paper
Pure Aspirin Powder
Watch Glass
Crude Aspirin Powder
Capillary Tube
Conc. H2SO4
Stirring Rod
Ethanol (96%)
Micro Pipette
Ethyl Acetate
200ml beaker
Distilled Water
500ml Volumetric Flask
37
PROCEDURE
1. Three 10cm x 20cm Aluminum TLC plates were obtained. A thin line was drawn 1cm from
2. Three chroma tanks were each filled with 10mL of a mixture of ethanol and ethyl acetate
in the ratios of
i. 7:3,
iii. 3:7 respectively. Each tank was then covered to allow the solvent enough time to
3. Very small amounts of the synthesised aspirin, the pure aspirin and the salicylic acid were
each dissolved in separate test–tubes using 1mL each of the 7:3 ethanol and ethyl acetate
mixture as prepared in step 2 (i) above. Each of the resulting solutions was then spotted
separately on the faint line on one of the plates using capillary tubes. The plate was allowed
to dry and then placed in the chromatank containing the 7:3 ethanol and ethyl acetate
mixture.
4. The chroma tank was covered with a glass tile and enough time allowed for the solvent to
elute.
6. The procedure was repeated for the 1:1 and 7:3 mixtures of the ethanol and ethyl acetate
prepared in steps 2(ii) and (iii) above using the other two TLC plates. The diameter of each
spot was ensured around 2mm and at equal intervals. Three spots were made on each TLC
plate, each spot representing the pure aspirin, the synthesised aspirin, and the salicylic acid.
7. A UV Lamp was used to view the spots which were carefully circled with pencil.
38
8. Their Rf values were determined.
3:7 8.5 - - - -
CALCULATION OF RF VALUES
i. Pure aspirin
𝟕.𝟖𝐜𝐦
Rf = 𝟗.𝟓𝐜𝐦 = 0.82
𝟕. 𝟔𝐜𝐦
Rf = = 0.80
𝟗. 𝟓𝐜𝐦
𝟗.𝟐𝐜𝐦
Rf = 𝟗.𝟓𝐜𝐦 = 0.97
39
𝟗. 𝟏𝐜𝐦
Rf = = 0.96
𝟗. 𝟓𝐜𝐦
i. Pure aspirin
𝟕.𝟏𝐜𝐦
Rf = 𝟗.𝟐𝐜𝐦 = 0.77
𝟕.𝟎𝐜𝐦
Rf = = 0.76
𝟗.𝟐𝐜𝐦
𝟖.𝟖𝐜𝐦
Rf = 𝟗.𝟐𝐜𝐦 = 0.96
𝟖.𝟖𝐜𝐦
Rf = 𝟗.𝟐𝐜𝐦 = 0.96
Rf =??
DISCUSSION
Three different mixtures of ethanol and ethyl acetate were used as solvents for the analysis in
i. 7:3
iii. 3:7
40
This was to help determine which of these solvent systems would be more efficient in the
For the 1:1 solvent system, the spot for the pure aspirin yielded an Rf value of 0.77 while the
spot for the synthesised aspirin yielded two separations with Rf values of 0.76 and 0.96. The 0.76
Rf value was tagged as x and the 0.96 Rf value was also tagged as y as can be seen from the table
above. Meanwhile the spotted salicylic acid also produced an Rf value of 0.96.
Comparing these Rf values shows that the separation tagged as y has the same Rf value as the
salicylic acid spot. This implies that separation y was highly likely to be salicylic acid which may
have been left in the synthesised aspirin probably due to inefficient washing and recrystallization.
This could also be due to decomposition of some of the synthesised aspirin, since it was kept for
some weeks before carrying out this analysis. Separation x could also be concluded to be the actual
aspirin synthesised judging from the proximity of its Rf values to that of the pure aspirin spot.
The same conclusions were arrived at using the Rf values for the 7:3 solvent system. The 3:7
solvent system produced no separations. No spots were identified after viewing under the UV lamp
using the 3:7 solvent system. Comparing the Rf values and separations obtained using the three
solvent systems implies that the 1:1 solvent system was the most efficient. The 1:1 solvent system
produced Rf values that were much closer to those of their respective standards (pure aspirin and
the pure salicylic acid) used. For example the Rf value for separation x was exactly the same as
that for the pure salicylic acid spot (0.96) using the 1:1 solvent system. The 7:3 solvent system
gave Rf values of 0.97 for separation x and 0.96 for the salicylic acid spot. The 3:7 solvent system
41
Ethanol is more polar than the ethyl acetate because of the presence of the OH group. In 3:7
mixture, the percentage of the ethanol in the mixture was quite small and so its polarity was
overcome by the relatively non-polar ethyl acetate hence the incidence of no separation.
CONCLUSION
The synthesised aspirin contained some amount impurity because of the presence of
42
REFERENCES
1. D.A. Skoog, D.M. West, F.J. Holler and S.R. Crouch, Fundamentals of Analytical
2. D.C. Harris, Quantitative Chemical Analysis, 7th ed. New York: W.H. Freeman and
Company, 2010.
3. R.A. Day, Jr. and A.L. Underwood, Quantitative Analysis, 6th ed. New Jersey:Prentice-
43
APPENDIX
1. 0.0010%
44
2. 0.00075%
3. 0.0015%
45
4. 0.00050%
46
5. SPECTRA FOR ALL CONCENTRATIONS OF STNADARD
47
APPENDIX B
48