Organic laboratory

Experiment: #4(B)
Due Date :19/2/2023

: Students Names
Tala Alzboon , Leen Aldibs
Afnan Abusrour , Abeer Ali , Salwa Ahed
Submitted to: Dr.Sami Makharza
:Introduction
Column chromatography is a method commonly used to separate
molecules in complex mixtures. Unlike planar chromatography, in
column chromatography, the stationary phase or resin is packed into a
column. The mobile phase is then passed through the packed
stationary phase to achieve separation. The purpose of this separation
can be analytical or preparative.
Both gas chromatography and liquid chromatography separations are
performed on columns. Our discussion here focuses on liquid column
chromatography systems.
Aim:
The aim of this experiment is to separate two substances using
column chromatography.

Materials: Conical flask burette

Ø Methylene blue (MW=373.90g/mol)

Ø Methyl orange (MW=327.34g/mol)
Ø Ethyl alcohol
Ø Water
Ø Silica powder graduated
burette
cylinder burette clamp stand
Experimental Procedure:
1) Assemble your column. Using a burette clamp, clamp the column to a stand. Put glass wool plug at
the bottom of column using a glass rod, then place a beaker below the tip of the column. Make sure
the column stopcock (drain valve) is closed.

2) Put 10 g of silica powder then 10 ml of ethanol and 10 g into the column, wash any silica powder
that sticks to the sides of the column with ethanol. Allow the silica to settle until you see 2-3 drops of
the solvent.

3) Add 5 drops of the green dye (methyl orange/methylene blue mixture) directly above the column
packing slowly, so that they do not splash up on the sides of the column. (If the do wash it with some
ethanol)

4) Carefully add ethanol (EtOH) down the side of the column until the solvent is 1-3 cm above the
level of the Silica. Open the stopcock and allow the eluent to drain into the beaker. While the (EtOH)
drains, notice that the dyes are starting to separate. Casually and (EtOH). Do not allow the solvent to
go below the level of silica.
5) When the blue dye is at the tip of the column.Replace the beaker with a 10 ml
graduated cylinder. Restart the flow and collect the column eluate until all of the
methyl orange has eluted (i.e., the yellow color disappears). Close the stopcock.
Record the volume in ml of methyl orange /EtOH collected.

6) Continue the process to Collect the blue dye (methylene blue). Measure and
record the volume in ml. (That part was not done)
Discussion:-
Different types of chromatography use various types of stationary and mobile phases. In
This experiment, the solid phase is silica gel while the mobile phase is an organic solvent
(Ethanol). This means that the compounds to be separated must choose between being
absorbed to the solid silica gel or moving along in the organic Solvent. The silica gel is either
packed into a column or adhered to a sheet of glass or plastic, Depending on the type of
chromatography.
When a column is used, as in this experiment, the compound mixture is placed on top and
the solvent are run down the column separating the mixture along the way. With the silica
gel on a plate, the compounds are placed close to the bottom and the mixture is separated
as the solvent travels up by means of capillary action. Since silica gel is a porous form of
SiO2, the surface of gel contains Si-OH and Si-O-Si functional groups. With silica gel, the
dominant interactive forces between the adsorbent and the materials to be separated are
of the dipole-Dipole type. Highly polar molecules interact fairly strongly with The polar Si—O
bonds of these adsorbents and will tend to stick or adsorb onto the fine particles of the
adsorbent while weakly polar molecules are held less tightly.
• For more clarification, let’s see these shapes:-
• Figure 1:- illustrates the General column chromatography set-up.

Silica
powder
• Figure 2: Illustration of a column chromatography separation of a mixture of 3 components,
• A, B and C. Compound A is the least polar and compound C is the most polar and the
• separation Occurs as solvent is allowed to flow through the stationary phase.
• ➔ In our experiment methyl orange is the least polar, and the methylene blue is the most polar
• blue is more polar than methyl orange and since silica gel is of highest
• polarity of all, Methylene blue will have a higher affinity for the stationary
• phase.
• Another factor that establishes the rate at which a compound travels past silica gel is the
• Polarity of the solvent. A polar solvent will compete for silica absorption sites, disallowing
• the Compounds to do so. This promotes a faster rate at which all compounds travel. The
• order in Which the compounds move remains the same, while moving faster as the polarity
• of the eluent (solvent system) increases.
• ➔ There for Chemistry- uses of ethanol. Ethanol is a very polar molecule due to its
• hydroxyl (OH) group, which forms hydrogen bonds with other molecules.
Conclusion:
• Column chromatography is described as the useful technique in which
the substances to be isolated are presented onto the highest point of
a column loaded with an adsorbent (stationary phase), go through the
column at various rates that rely upon the affinity of every substance
for the adsorbent and the solvent or solvent mixture, and are typically
gathered in solution as they pass from the column at various time.The
main principle involved in column chromatography is the adsorption
of the solutes of the solution with the help of a stationary phase and
afterward separates the mixture into independent components.