Mohammed Salam Unit 2 LA C
Separate to identify
Introduction:
Chromatography is commonly used for separating a mixture of the chemical substances. When the
mixture is separated the components are then analysed. Chromatography is commonly used in the
laboratories where chemist and biologists are aware of the principles of chromatography.
Chromatography was used to separate the pigments of chlorophyll in the experiment, Both TLC and
Paper chromatography was used to view and calculate the separation of the pigments. As the
solvent was moved along the paper and TLC it created smudges and was separated from its original
place. These spots or smudges have assorted colours and they determine the identification of
components of the mixture. So, the TLC and Paper chromatography is the stationary phase, and the
solvent is the mobile phase.
Capillary action operates primarily when the stationary phase (polar inorganic solvent water) and
mobile phase (nonpolar organic solvents) interact in a paper chromatographic system.
Thin layer chromatography (TLC) is composed of silica gel or alumina coated onto a piece of glass,
metal or plastic.
There are two types of phases:
Mobile phase – where it moves, it is usually a solvent or mixture of solvent.
Stationary phase – phase in chromatography that does not allow to move like paper and TLC.
How it works?
Here are the steps of how it works:
As the mobile phase stream runs continuously, an analyte is delivered at a precise amount.
In the mobile phase, analytes are carried to and through the stationary phase.
When the analyte reaches the stationary phase, the components of the analyte selectively
interact with the stationary phase, with some components interacting more and others
interacting less, resulting in the desired separation.
During separation, the analyte components are carried by the mobile phase to a special
instrument, which detects their presence in the mobile phase and generates a signal that
can be analysed for the separation.
In the experiment, the chromatography of amino acids was also used to determine the presence of
different protein. Each protein will have different Rf values due to having different R groups or
variable groups which makes them to have different properties. Paper chromatography is useful
when determining the distinct types of amino acids.
Aspartame, a cancer-causing chemical formed by a mold on peanuts, can also be detected and
separated using chromatography in the food business. Various additives, vitamins, preservatives,
and proteins can also be detected using chromatography.
We used paper chromatography in amino acids instead of TLC paper as amino acid does show in TLC
this is due to ninhydrin spray cannot be detected in the tlc paper this is due to amino acids are
colourless, so ninhydrin spray is used to reveal the amino spots.
Chromatography of chlorophyll
1
, Mohammed Salam Unit 2 LA C
Paper chromatography of chlorophyll
Equipment:
Mortar
Chromatography paper
Pencil
Ruler
Capillary tubes
Leaf (spinach)
Propanone
Test tube
Scissors
Sellotape
Preparing for plant solution:
Before commencing the experiment, plant solution is required for the experiment for paper
chromatography. Used a scissor to cut up the spinach leaves into smaller parts so that it can be easy
to crush up the leaves. Mortar and pestle were used to crush the leaf so that it can be used to
extract the chlorophyll when using it with the solvents. To make it easier the leaf has been cut off
into smaller pieces with scissors so that it can be easier to make the solution. Small amount of
propanone also known as acetone was used in the plant solution. Acetone propanone dissolves
chlorophyll by breaking down its lipid bonds to a plant's thylakoid structure.
Fluids can adhere to capillary tubes in both vertical and horizontal directions and can therefore
move through them. Capillarity is the movement of a liquid through a small cross-section tube like
the openings in porous materials. The fluid is propelled upwards by capillary action. Capillary tubes
are composed of glass or plastic they are mostly used in chromatography. Scientists and biologists
commonly use glass capillary tubes for experiments that need extremely small samples, the most
common type being glass capillary tubes. Capillary tube are used in chromatography paper and TLC
this is due to the small volume contained in the tube. To test for a liquid, a capillary action is used to
draw it into a glass tube, followed by contact with a TLC plate or chromatography paper to draw it
out.
Method:
Used a ruler to draw a line on the chromatography paper the line should be above 1cm so that after
putting it into the beaker with propanone it should not touch the spot immediately, the capillary
tube was used to transport the plant solution onto the centre of the line (stationary phase). The line
that was drawn on the chromatography was pencil this is because pencil is insoluble so it would not
produce error. It was then left to dry and was repeated for five times on the same spot to ensure
that the area was highly concentrated.
After that, a small amount of propanone was applied in the beaker and the paper was hung so that it
was dipped in the propanone. Before putting the paper into the propanone, the chromatography
paper was sticked with a wooden stick and Sellotape above the paper, this is required as it would
reduce experimental error. The paper was then placed into the beaker and made sure it was hung so
that the propanone was below the spot. The paper chromatography was then left in the beaker until
the propanone had soaked in near the top. The paper chromatography was then taken out of the
beaker and drew a line with a pencil where the propanone has reached. A pencil was also drawn
2
Separate to identify
Introduction:
Chromatography is commonly used for separating a mixture of the chemical substances. When the
mixture is separated the components are then analysed. Chromatography is commonly used in the
laboratories where chemist and biologists are aware of the principles of chromatography.
Chromatography was used to separate the pigments of chlorophyll in the experiment, Both TLC and
Paper chromatography was used to view and calculate the separation of the pigments. As the
solvent was moved along the paper and TLC it created smudges and was separated from its original
place. These spots or smudges have assorted colours and they determine the identification of
components of the mixture. So, the TLC and Paper chromatography is the stationary phase, and the
solvent is the mobile phase.
Capillary action operates primarily when the stationary phase (polar inorganic solvent water) and
mobile phase (nonpolar organic solvents) interact in a paper chromatographic system.
Thin layer chromatography (TLC) is composed of silica gel or alumina coated onto a piece of glass,
metal or plastic.
There are two types of phases:
Mobile phase – where it moves, it is usually a solvent or mixture of solvent.
Stationary phase – phase in chromatography that does not allow to move like paper and TLC.
How it works?
Here are the steps of how it works:
As the mobile phase stream runs continuously, an analyte is delivered at a precise amount.
In the mobile phase, analytes are carried to and through the stationary phase.
When the analyte reaches the stationary phase, the components of the analyte selectively
interact with the stationary phase, with some components interacting more and others
interacting less, resulting in the desired separation.
During separation, the analyte components are carried by the mobile phase to a special
instrument, which detects their presence in the mobile phase and generates a signal that
can be analysed for the separation.
In the experiment, the chromatography of amino acids was also used to determine the presence of
different protein. Each protein will have different Rf values due to having different R groups or
variable groups which makes them to have different properties. Paper chromatography is useful
when determining the distinct types of amino acids.
Aspartame, a cancer-causing chemical formed by a mold on peanuts, can also be detected and
separated using chromatography in the food business. Various additives, vitamins, preservatives,
and proteins can also be detected using chromatography.
We used paper chromatography in amino acids instead of TLC paper as amino acid does show in TLC
this is due to ninhydrin spray cannot be detected in the tlc paper this is due to amino acids are
colourless, so ninhydrin spray is used to reveal the amino spots.
Chromatography of chlorophyll
1
, Mohammed Salam Unit 2 LA C
Paper chromatography of chlorophyll
Equipment:
Mortar
Chromatography paper
Pencil
Ruler
Capillary tubes
Leaf (spinach)
Propanone
Test tube
Scissors
Sellotape
Preparing for plant solution:
Before commencing the experiment, plant solution is required for the experiment for paper
chromatography. Used a scissor to cut up the spinach leaves into smaller parts so that it can be easy
to crush up the leaves. Mortar and pestle were used to crush the leaf so that it can be used to
extract the chlorophyll when using it with the solvents. To make it easier the leaf has been cut off
into smaller pieces with scissors so that it can be easier to make the solution. Small amount of
propanone also known as acetone was used in the plant solution. Acetone propanone dissolves
chlorophyll by breaking down its lipid bonds to a plant's thylakoid structure.
Fluids can adhere to capillary tubes in both vertical and horizontal directions and can therefore
move through them. Capillarity is the movement of a liquid through a small cross-section tube like
the openings in porous materials. The fluid is propelled upwards by capillary action. Capillary tubes
are composed of glass or plastic they are mostly used in chromatography. Scientists and biologists
commonly use glass capillary tubes for experiments that need extremely small samples, the most
common type being glass capillary tubes. Capillary tube are used in chromatography paper and TLC
this is due to the small volume contained in the tube. To test for a liquid, a capillary action is used to
draw it into a glass tube, followed by contact with a TLC plate or chromatography paper to draw it
out.
Method:
Used a ruler to draw a line on the chromatography paper the line should be above 1cm so that after
putting it into the beaker with propanone it should not touch the spot immediately, the capillary
tube was used to transport the plant solution onto the centre of the line (stationary phase). The line
that was drawn on the chromatography was pencil this is because pencil is insoluble so it would not
produce error. It was then left to dry and was repeated for five times on the same spot to ensure
that the area was highly concentrated.
After that, a small amount of propanone was applied in the beaker and the paper was hung so that it
was dipped in the propanone. Before putting the paper into the propanone, the chromatography
paper was sticked with a wooden stick and Sellotape above the paper, this is required as it would
reduce experimental error. The paper was then placed into the beaker and made sure it was hung so
that the propanone was below the spot. The paper chromatography was then left in the beaker until
the propanone had soaked in near the top. The paper chromatography was then taken out of the
beaker and drew a line with a pencil where the propanone has reached. A pencil was also drawn
2
