CHROMATOGRAPHY WITH PLANT EXTRACTS AND AMINO ACIDS
In my role as a Technical Assistant for Chemical Equip, I was given the task of using
chromatography to determine the retention factor. There are many industrial applications of
chromatography, places such as the food industry where these techniques are used to
analyse food products for things such as additives, proteins etc. pharmaceutical industries
where samples are analysed to check the clarity, forensic labs where samples from an
allocated crime scene are analysed and in the chemical industries where chromatography is
used as a variety of purposes, requiring the separation and identity of a chemical substance.
Paper chromatography uses paper (coffee filter) as the stationary phase, the type of paper
chromatography is very important, because other types of paper can either keep the results
the same or can drastically alter the values found, for example, normal paper (used to
usually write with) is completely coated to avoid ink from bleeding into the page will writing,
in turn making this type of paperless likely to receive any satisfactory results. Similarly, wax
paper, which is not absorbent, would make it almost impossible to gain any satisfactory
results. [1]Paper chromatography is an analytical method used to separate coloured
chemicals or substances. Out of paper, TLC and amino, primarily paper is the easiest
method of thin-layer chromatography whereas, thin-layer chromatography could be slightly
more challenging as touching the material can easily break it off. Most chromatography
involves two solvents, along with an extract/solvent. This is used for separating complex
mixtures (in amino acids etc.) The mobile phase is the solvent that ascends the stationary
phase. As the solvent moves up the paper, the sample becomes dissolved in it and begins to
travel p the chromatography paper. The molecules in the sample move at different rates
because they differ in how soluble they are, their molecular mass and how they bond with
the paper being used. Larger molecules will move a lot slower than a smaller molecule, so
separation is achieved according to the size of the molecule, the more soluble a component,
the further it will move along the paper. The mobile phase overall is a mixture of non-polar
solvents, and stationery is a polar solvent. The difference between thin-layer
chromatography is that the stationary phase in TLC is a layer of usual silica or aluminium
oxide, and stationary in the paper, in this case, propane. Thin-layer chromatography has the
same technique; however, we use a silica-coated plate instead of filter paper, the silica we
use for chromatography is very polar because silica contains OH groups (this group is
attached to anything related to the hydroxyl group, which means that it attaches to some
molecules containing oxygen and hydrogen, bonded with one another), because of this
protein compounds and hydrogen bonds are formed. Compounds that are a lot more
attracted to by the silica stick more firmly in the stationary phase and travel up as far up the
paper in the mobile phase. These techniques are useful because if a company needed to
identify the unknown sample, we could compare them to published results and against our
results to identify the unknown samples. The separation of the components can be
identified by calculating the Rf value. The retention factor values are calculated by
measuring the points where each component of the compound has separated from other
components and stopped moving along the paper or TLC plate. Once molecules have been
separated the different components can be identified by finding the Rf value. Rf is calculated
using the formula:
, Rf= distance moved by the compound/distance moved by the solvent
EQUIPMENT FOR SIMPLE PAPER AND TLC PAPER
● Spinach (plant extract)
● Propanone
● 3-cylinder glass tubes
● Rack
● Filter paper (simple paper) or silica plate (thin-layer)
● Capillary tube
● Pestle and mortar
● Pipette
● Pencil
● Ruler
● Tweezers
● Gloves
● Fume cupboard
EQUIPMENT FOR AMINO ACIDS
● Propanone
● Phenylalanine acid
● Proline acid
● Serine acid
● Filter paper
● Pencil
● Ruler
● Ninhydrin spray
● Tweezers
● Gloves
● Cylinder glass tubes
● Capillary tubes
● Pipette
● Rack
● Oven/ hairdryer
● Beaker
● Scissors
● Fume cupboard
In my role as a Technical Assistant for Chemical Equip, I was given the task of using
chromatography to determine the retention factor. There are many industrial applications of
chromatography, places such as the food industry where these techniques are used to
analyse food products for things such as additives, proteins etc. pharmaceutical industries
where samples are analysed to check the clarity, forensic labs where samples from an
allocated crime scene are analysed and in the chemical industries where chromatography is
used as a variety of purposes, requiring the separation and identity of a chemical substance.
Paper chromatography uses paper (coffee filter) as the stationary phase, the type of paper
chromatography is very important, because other types of paper can either keep the results
the same or can drastically alter the values found, for example, normal paper (used to
usually write with) is completely coated to avoid ink from bleeding into the page will writing,
in turn making this type of paperless likely to receive any satisfactory results. Similarly, wax
paper, which is not absorbent, would make it almost impossible to gain any satisfactory
results. [1]Paper chromatography is an analytical method used to separate coloured
chemicals or substances. Out of paper, TLC and amino, primarily paper is the easiest
method of thin-layer chromatography whereas, thin-layer chromatography could be slightly
more challenging as touching the material can easily break it off. Most chromatography
involves two solvents, along with an extract/solvent. This is used for separating complex
mixtures (in amino acids etc.) The mobile phase is the solvent that ascends the stationary
phase. As the solvent moves up the paper, the sample becomes dissolved in it and begins to
travel p the chromatography paper. The molecules in the sample move at different rates
because they differ in how soluble they are, their molecular mass and how they bond with
the paper being used. Larger molecules will move a lot slower than a smaller molecule, so
separation is achieved according to the size of the molecule, the more soluble a component,
the further it will move along the paper. The mobile phase overall is a mixture of non-polar
solvents, and stationery is a polar solvent. The difference between thin-layer
chromatography is that the stationary phase in TLC is a layer of usual silica or aluminium
oxide, and stationary in the paper, in this case, propane. Thin-layer chromatography has the
same technique; however, we use a silica-coated plate instead of filter paper, the silica we
use for chromatography is very polar because silica contains OH groups (this group is
attached to anything related to the hydroxyl group, which means that it attaches to some
molecules containing oxygen and hydrogen, bonded with one another), because of this
protein compounds and hydrogen bonds are formed. Compounds that are a lot more
attracted to by the silica stick more firmly in the stationary phase and travel up as far up the
paper in the mobile phase. These techniques are useful because if a company needed to
identify the unknown sample, we could compare them to published results and against our
results to identify the unknown samples. The separation of the components can be
identified by calculating the Rf value. The retention factor values are calculated by
measuring the points where each component of the compound has separated from other
components and stopped moving along the paper or TLC plate. Once molecules have been
separated the different components can be identified by finding the Rf value. Rf is calculated
using the formula:
, Rf= distance moved by the compound/distance moved by the solvent
EQUIPMENT FOR SIMPLE PAPER AND TLC PAPER
● Spinach (plant extract)
● Propanone
● 3-cylinder glass tubes
● Rack
● Filter paper (simple paper) or silica plate (thin-layer)
● Capillary tube
● Pestle and mortar
● Pipette
● Pencil
● Ruler
● Tweezers
● Gloves
● Fume cupboard
EQUIPMENT FOR AMINO ACIDS
● Propanone
● Phenylalanine acid
● Proline acid
● Serine acid
● Filter paper
● Pencil
● Ruler
● Ninhydrin spray
● Tweezers
● Gloves
● Cylinder glass tubes
● Capillary tubes
● Pipette
● Rack
● Oven/ hairdryer
● Beaker
● Scissors
● Fume cupboard
