Hebron University

Faculty of Pharmacy of Medical Sciences

Introduction to Medical Concepts

Midterm exam
Due date: 9-4-2017

Name:.. Instructor:.

Q1: Re-write the following procedure as it should be in your lab report.

Procedure:
1. Cut TLC plates. Usually silica plates are bought as square glass pieces that must be cut using a
diamond tipped glass cutter and following a template. Before scoring the glass, use a ruler and a pencil
to lightly mark baselines on the silica side of the plate (be careful not to remove any silica from the
plate). Using a sharp glass cutter and a ruler as a guide, you should have no problem scoring the glass.
Once the entire plate is scored, you can then break the glass into individual pieces. (In the beginning
this may be frustrating, but after some practice, you should become comfortable with this technique.)
Baselines of the silica plate was marked using a pencil and ruler, the marked baselines was cut using a
sharp glass cutter, the a desired template was broken gently.

2. Determine the solvent system. Your compounds will travel different distances up the plate depending
on the solvent you choose. In non-polar solvents like pentane and hexane, most polar compounds will
not move, while non-polar compounds will travel some distance up the plate. In contrast, polar solvents
will usually move non-polar compounds to the solvent front and push the polar compounds off of the
baseline. A good solvent system is one that moves all components of your mixture off the baseline, but
does not put anything on the solvent front - Rf values between 0.15 and 0.85. This is not always
possible, but should be your goal when running a TLC. (For column chromatography the correct
solvent system should give an Rf between 0.2 and 0.3.) Now, which solvents to pick? Here's a list of
some standard solvents and their polarity (from LLP):

Very polar additives:

Methanol > Ethanol > Isopropanol
Moderately polar additives:
Acetonitrile > Ethyl Acetate > Chloroform >
Dichloromethane > Diethyl Ether > Toluene
Non-polar additives:
Cyclohexane, Petroleum Ether, Hexane, Pentane
The suitable solvents: Mixed solvents of Ethyl Acetate and hexane.

3. Fill TLC chamber with 1-2 mL of the desired solvent system. Place a large piece of cut filter paper in
the chamber as well.

The mixed solvents of Ethyl acetate and hexane were added to the chamber containing a large
filer paper.

1
To be filled electronically, printed on paper, and handed to your instructor.
 Hebron University
Faculty of Pharmacy of Medical Sciences

Introduction to Medical Concepts

Midterm exam
Due date: 9-4-2017
4. Spot the compound on the baseline of the TLC plate. We will use commercial spotters, but spotters
can be pulled from hot Pasteur pipets. If you are monitoring a reaction, make sure to spot the starting
material, the reaction mixture, and a co-spot of both.
The compounds were spotted on the baseline using a pipet.

5. Run the TLC. Let the solvent go about 90% of the way up the plate.
The solvent was waited until reached the way up the plate.
6. Remove the plate from the chamber and mark the solvent front immediately with a pencil - you
will use this to calculate the Rf.
The plate was removed from chamber and the solvent front was marked immediately.

7. Let the solvent dry off of the plate.

The solvent was let to dry off the plate.

8. Visualize the TLC using non-destructive technique(s). The best non-destructive method is the UV
lamp. Place your plate under the UV lamp and circle any UV active spots with your pencil.
The plate was placed under the UV lamp and UV active spots were circled.

Q2: Identify mistakes in the following results section.

Results 1. The first sentence

The aim of this experiment was to learn how to separate compounds should be inon
based the
their
introduction section
relative polarity. The compounds we used are listed in the in the following table:
not in result section.
1 Aniline 2. The table is not
2 4-nitroaniline needed, sentences
3 Aniline+ 4-nitroaniline would be enough.
3. If the table is needed
Each one of them was soaked in the solvent and allowed to move it up the TLC
should not beplate.
We want to compare distances traveled on the plates related to one another
placed hereand the
, it should
question was asked why it is important to quantify our results. beThis below.
problem and
4. The
question were solved by looking the total distance traveled by the solutionfirst sentence
and the
under the table must
distance covered by the compound placed towards the bottom of the paper. After
be in method section
looking at it, it was determined that if one was to take the 5. distance traveled and
The question and the
divide it by the total distance the solution in this case the eluent traveled
answer mustup bethe
in
paper it would give a value. This value is called the Rf value and is a
discussion section.
compare the distances traveled on the plate. 6. in the figure, the

2
To be filled electronically, printed on paper, and handed to your instructor.
 Hebron University
Faculty of Pharmacy of Medical Sciences

Introduction to Medical Concepts

Midterm exam
Due date: 9-4-2017
figure is not given a
Substances can be identified by the heights they reach on specific
the name for the
completed
compounds,
chromatogram by calculating Rf (rate of flow or retention factor) values. The Rand it
value
is a constant for a given substance under the same experimentalshould be given
conditions. The aRf
number (1)
value may be calculated from the following equation. The Rf
7. In the figure (1+2)
should be replaced by
3 as mentioned in the
table above ( the
The following figure shows the distances travelled by the solvent
mixture).and the 3
substances
8. In the sentence below
the figure, the phrase
(the figure shows) is
wrong, the correction
is to delete this
phrase and begin with
distance. (figure
1).

9. In the first sentence

the results not given a
specific unit (cm, mm,
m).

10. The results also

The figure shows that the distance travelled by aniline is 6, nitroaniline is 7.5, in
are written and by
present
the mixture is 3.2. tense which is
completely wrong it
11 cm was the du=distance travelled by the solvent itself. should be in the past
0 tense, example (the
The following diagram shows the 4 distances as compared to eachdistance
other: travelled by
aniline was 6 cm).

11. There is a
mistake in the second
sentence (du=) it
should be deleted.

3
To be filled electronically, printed on paper, and handed to your instructor.
 Hebron University
Faculty of Pharmacy of Medical Sciences

Introduction to Medical Concepts

Midterm exam
Due date: 9-4-2017

As we see from the figures, as the polarity of the compound increased the distance
travelled decreased. This can be explained based on the polar-polar interaction
principle.
Calculation of Rf:

We calculated the Rf values as follows:

Distance travelled by aniline 6
Rf for aniline= = =0.545
Distance travelled by solvent 11

Distancetravelled by 4nitroaniline 7.5

Rf for 4nitroaniline= = =0.681
Distance travelled by solvent 11

Distance travelled by mixture 3.2

Rf for the mixture= = =0.29
Distance travelled by solvent 11

The Rf values are arranged in the following table:

Aniline 0.545
4-nitroaniline 0.681
Aniline + 4-nitroaniline 0.29

As we can see the Rf value increased as the polarity of the substance increased and
this can be used for various indications.

4
To be filled electronically, printed on paper, and handed to your instructor.
 Hebron University
Faculty of Pharmacy of Medical Sciences

Introduction to Medical Concepts

Midterm exam
Due date: 9-4-2017

EXTRA NOTES:

1. The figure in the last page, is not given a specific name and also not
labeled (should be figure 2)
2. The sentence below the figure (As we see from the figures, as the polarity of
the compound increased the distance travelled decreased. This can be explained
based on the polar-polar interaction principle) should be in discussion section
(without as we see from figure)
3. The figure also mistaken (1+2) label should be replaced by 3 as
mentioned in the table above.
4. (we calculated the Rf values as follows) wrong, ( Rf values were
calculated as followed) correct
5. in determining the Rf values the equation is not needed to be repeated
again and again it is mentioned above.
6. The table is not needed sentences would be enough to present that like
(the Rf value of aniline was determined to be 0.545 (this is also applied
to nitroaniline and mixture).
7. The last sentence should be in discussion section (as we can see the Rf
value increased as the polarity of the substance increased and this can be used
for various indications.) but it should be like this (Rf value increased as the
polarity of the substance increased and this can be used for various indications
(figure 2)).
8. The axises in the second figure are not labeled (polarity, distance).

5
To be filled electronically, printed on paper, and handed to your instructor.