paper chromatography biology experiment

Chromatography is used to separate mixtures of substances into their components. All forms of chromatography work on the same principle.

They all have a (a solid, or a liquid supported on a solid) and a (a liquid or a gas). The mobile phase flows through the stationary phase and carries the components of the mixture with it. Different components travel at different rates. We'll look at the reasons for this further down the page.

In paper chromatography, the stationary phase is a very uniform absorbent paper. The mobile phase is a suitable liquid solvent or mixture of solvents.

You probably used paper chromatography as one of the first things you ever did in chemistry to separate out mixtures of coloured dyes - for example, the dyes which make up a particular ink. That's an easy example to take, so let's start from there.

Suppose you have three blue pens and you want to find out which one was used to write a message. Samples of each ink are spotted on to a pencil line drawn on a sheet of chromatography paper. Some of the ink from the message is dissolved in the minimum possible amount of a suitable solvent, and that is also spotted onto the same line. In the diagram, the pens are labelled 1, 2 and 3, and the message ink as M.

The reason for covering the container is to make sure that the atmosphere in the beaker is saturated with solvent vapour. Saturating the atmosphere in the beaker with vapour stops the solvent from evaporating as it rises up the paper.

As the solvent slowly travels up the paper, the different components of the ink mixtures travel at different rates and the mixtures are separated into different coloured spots.

The diagram shows what the plate might look like after the solvent has moved almost to the top.

It is fairly easy to see from the final chromatogram that the pen that wrote the message contained the same dyes as pen 2. You can also see that pen 1 contains a mixture of two different blue dyes - one of which be the same as the single dye in pen 3.

values

Some compounds in a mixture travel almost as far as the solvent does; some stay much closer to the base line. The distance travelled relative to the solvent is a constant for a particular compound as long as you keep everything else constant - the type of paper and the exact composition of the solvent, for example.

The distance travelled relative to the solvent is called the R value. For each compound it can be worked out using the formula:

For example, if one component of a mixture travelled 9.6 cm from the base line while the solvent had travelled 12.0 cm, then the R value for that component is:

In the example we looked at with the various pens, it wasn't necessary to measure R values because you are making a direct comparison just by looking at the chromatogram.

You are making the assumption that if you have two spots in the final chromatogram which are the same colour and have travelled the same distance up the paper, they are most likely the same compound. It isn't necessarily true of course - you could have two similarly coloured compounds with very similar R values. We'll look at how you can get around that problem further down the page.

In some cases, it may be possible to make the spots visible by reacting them with something which produces a coloured product. A good example of this is in chromatograms produced from amino acid mixtures.

Suppose you had a mixture of amino acids and wanted to find out which particular amino acids the mixture contained. For simplicity we'll assume that you know the mixture can only possibly contain five of the common amino acids.

A small drop of a solution of the mixture is placed on the base line of the paper, and similar small spots of the known amino acids are placed alongside it. The paper is then stood in a suitable solvent and left to develop as before. In the diagram, the mixture is M, and the known amino acids are labelled 1 to 5.

The position of the solvent front is marked in pencil and the chromatogram is allowed to dry and is then sprayed with a solution of . Ninhydrin reacts with amino acids to give coloured compounds, mainly brown or purple.

The left-hand diagram shows the paper after the solvent front has almost reached the top. The spots are still invisible. The second diagram shows what it might look like after spraying with ninhydrin.

There is no need to measure the R values because you can easily compare the spots in the mixture with those of the known amino acids - both from their positions and their colours.

In this example, the mixture contains the amino acids labelled as 1, 4 and 5.

And what if the mixture contained amino acids other than the ones we have used for comparison? There would be spots in the mixture which didn't match those from the known amino acids. You would have to re-run the experiment using other amino acids for comparison.

Two way paper chromatography gets around the problem of separating out substances which have very similar R values.

I'm going to go back to talking about coloured compounds because it is much easier to see what is happening. You can perfectly well do this with colourless compounds - but you have to use quite a lot of imagination in the explanation of what is going on!

This time a chromatogram is made starting from a single spot of mixture placed towards one end of the base line. It is stood in a solvent as before and left until the solvent front gets close to the top of the paper.

In the diagram, the position of the solvent front is marked in pencil before the paper dries out. This is labelled as SF1 - the solvent front for the first solvent. We shall be using two different solvents.

If you look closely, you may be able to see that the large central spot in the chromatogram is partly blue and partly green. Two dyes in the mixture have almost the same R values. They could equally well, of course, both have been the same colour - in which case you couldn't tell whether there was one or more dye present in that spot.

What you do now is to wait for the paper to dry out completely, and then rotate it through 90°, and develop the chromatogram again in a different solvent.

It is very unlikely that the two confusing spots will have the same R values in the second solvent as well as the first, and so the spots will move by a different amount.

The next diagram shows what might happen to the various spots on the original chromatogram. The position of the second solvent front is also marked.

You wouldn't, of course, see these spots in both their original and final positions - they have moved! The final chromatogram would look like this:

Two way chromatography has completely separated out the mixture into four distinct spots.

If you want to identify the spots in the mixture, you obviously can't do it with comparison substances on the same chromatogram as we looked at earlier with the pens or amino acids examples. You would end up with a meaningless mess of spots.

You can, though, work out the R values for each of the spots in both solvents, and then compare these with values that you have measured for known compounds under exactly the same conditions.

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Paper is made of cellulose fibres, and cellulose is a polymer of the simple sugar, glucose.

The key point about cellulose is that the polymer chains have -OH groups sticking out all around them. To that extent, it presents the same sort of surface as silica gel or alumina in thin layer chromatography.

It would be tempting to try to explain paper chromatography in terms of the way that different compounds are adsorbed to different extents on to the paper surface. In other words, it would be nice to be able to use the same explanation for both thin layer and paper chromatography. Unfortunately, it is more complicated than that!

The complication arises because the cellulose fibres attract water vapour from the atmosphere as well as any water that was present when the paper was made. You can therefore think of paper as being cellulose fibres with a very thin layer of water molecules bound to the surface.

It is the interaction with this water which is the most important effect during paper chromatography.

Suppose you use a non-polar solvent such as hexane to develop your chromatogram.

Non-polar molecules in the mixture that you are trying to separate will have little attraction for the water molecules attached to the cellulose, and so will spend most of their time dissolved in the moving solvent. Molecules like this will therefore travel a long way up the paper carried by the solvent. They will have relatively high R values.

On the other hand, polar molecules have a high attraction for the water molecules and much less for the non-polar solvent. They will therefore tend to dissolve in the thin layer of water around the cellulose fibres much more than in the moving solvent.

Because they spend more time dissolved in the stationary phase and less time in the mobile phase, they aren't going to travel very fast up the paper.

The tendency for a compound to divide its time between two immiscible solvents (solvents such as hexane and water which won't mix) is known as . Paper chromatography using a non-polar solvent is therefore a type of .

A moment's thought will tell you that partition can't be the explanation if you are using water as the solvent for your mixture. If you have water as the mobile phase and the water bound on to the cellulose as the stationary phase, there can't be any meaningful difference between the amount of time a substance spends in solution in either of them. All substances should be equally soluble (or equally insoluble) in both.

And yet the first chromatograms that you made were probably of inks using water as your solvent.

If water works as the mobile phase as well being the stationary phase, there has to be some quite different mechanism at work - and that must be equally true for other polar solvents like the alcohols, for example. Partition only happens between solvents which don't mix with each other. Polar solvents like the small alcohols do mix with water.

In researching this topic, I haven't found any easy explanation for what happens in these cases. Most sources ignore the problem altogether and just quote the partition explanation without making any allowance for the type of solvent you are using. Other sources quote mechanisms which have so many strands to them that they are far too complicated for this introductory level. I'm therefore not taking this any further - you shouldn't need to worry about this at UK A level, or its various equivalents.

Where would you like to go now?

To the chromatography menu . . .

To the analysis menu . . .

To Main Menu . . .

© Jim Clark 2007 (modified July 2016)

Chromatography ( OCR A Level Biology )

Revision note.

Biology Lead

Chromatography

• Chromatography is a technique that can be used to separate a mixture into its individual components
• Chromatography relies on differences in the solubility of the different chemicals (called ‘solutes’ ) within a mixture
• The mobile phase
• The stationary phase
• The components in the mixture separate as the mobile phase travels over the stationary phase
• Differences in the solubility of each component in the mobile phase affects how far each component can travel
• Those components with higher solubility will travel further than the others
• This is because they spend more time in the mobile phase and are thus carried further up the paper than the less soluble components

Paper chromatography

• Paper chromatography is one specific form of chromatography
• The mobile phase is the solvent in which the sample molecules can move, which in paper chromatography is a liquid e.g. water or ethanol
• The stationary phase in paper chromatography is the chromatography paper

Paper chromatography method

• A spot of the mixture (that you want to separate) is placed on chromatography paper and left to dry
• The chromatography paper is then suspended in a solvent
• Larger molecules move slower than smaller ones
• This causes the original mixture to separate out into different spots or bands on the chromatography paper
• This produces what is known as a chromatogram

An example of a chromatogram that has been produced by using paper chromatography to separate a spot of ink

Using chromatography to separate a mixture of monosaccharides

• Paper chromatography can be used to separate a mixture of monosaccharides
• Mixtures containing coloured molecules, such as ink or chlorophyll, do not have to be stained as they are already coloured
• Mixtures of colourless molecules, such as a mixture of monosaccharides, have to be stained first
• A spot of the stained monosaccharide sample mixture is placed on a line at the bottom of the chromatography paper
• Spots of known standard solutions of different monosaccharides are then placed on the line beside the sample spot
• As the solvent travels up through the chromatography paper, the different monosaccharides within the mixture separate out at different distances from the line
• If a spot from the monosaccharide sample mixture is at the same distance from the line as a spot from one of the known standard solutions, then the mixture must contain this monosaccharide

How chromatography can be used to separate a mixture of monosaccharides and identify the individual components

Using chromatography to separate a mixture of amino acids

• Paper chromatography can be used to separate a mixture of amino acids
• A spot of the unknown amino acid sample mixture is placed on a line at the bottom of the chromatography paper
• Spots of known standard solutions of different amino acids are then placed on the line beside the unknown sample spot
• If a spot from the amino acid sample mixture is at the same distance from the line as a spot from one the known standard solutions, then the mixture must contain this amino acid
• In order to view the spots from the different amino acids, it may be necessary to first dry the chromatography paper and then spray it with ninhydrin solution (this chemical reacts with amino acids , producing an easily visible blue-violet colour )

How chromatography can be used to separate a mixture of amino acids and identify the individual components

Calculating the Rf value

• After a chromatogram has been obtained the molecules present in the sample mixture can be identified by calculating their retardation factor (Rf)
• This line is known as the solvent front
• The distance between the origin line and the solvent front is the distance moved by the solvent
• The origin line is the line at the bottom of the paper on which the samples were placed at the beginning of the experiment
• A smaller R f value indicates the molecule is less soluble and larger in size
• The Rf value of each solute (each spot on the chromatogram) is calculated and then compared to the Rf values of known molecules/substances
• The equation is:
• It has no units

Using Rf values to identify chloroplast pigments

• Chromatography can be used to separate and identify chloroplast pigments that have been extracted from a leaf as each pigment will have a unique Rf value
• Carotenoids have the highest Rf values (usually close to 1)
• Chlorophyll B  has a much lower Rf value
• Chlorophyll A has an Rf value somewhere between those of carotenoids and chlorophyll B
• Small Rf values indicate the pigment is less soluble and larger in size

Paper chromatography can be used to separate the photosynthetic pigments found within chloroplasts. Rf values can then be calculated for each pigment and compared to known Rf values for the different pigments.

It is always worth trying to understand why  a certain practical technique is useful. An example of when chromatography would be used is if you have an unknown liquid and you have determined it contains protein using a Biuret test. Chromatography will then show you which amino acids are present so you can better understand the potential use or function of the sample. This could be useful in crime scene investigations or in detecting additives or spoilage in foods.

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Lára graduated from Oxford University in Biological Sciences and has now been a science tutor working in the UK for several years. Lára has a particular interest in the area of infectious disease and epidemiology, and enjoys creating original educational materials that develop confidence and facilitate learning.

Separation of Plant Pigments by Paper Chromatography

The separation of plant pigments by paper chromatography is an analysis of pigment molecules of the given plant. Chromatography refers to colour writing . This method separates molecules based on size, density and absorption capacity.

Chromatography depends upon absorption and capillarity . The absorbent paper holds the substance by absorption. Capillarity pulls the substance up the absorbent medium at different rates.

Separated pigments show up as coloured streaks . In paper chromatography, the coloured bands separate on the absorbent paper. Chlorophylls, anthocyanins, carotenoids, and betalains are the four plant pigments.

This post discusses the steps of separating plant pigments through paper chromatography. Also, you will get to know the observation table and the calculation of the Rf value.

Content: Separation of Plant Pigments by Paper Chromatography

Paper chromatography, plant pigments, steps of plant pigment separation, observation, calculation.

It is the simplest chromatography method given by Christian Friedrich Schonbein in 1865. Paper chromatography uses filter paper with uniform porosity and high resolution.

The mixtures in compounds have different solubilities . For this reason, they get separated distinctly between the stationary and running phase.

• The mobile phase is a combination of non-polar organic solvents. The solvent runs up the stationary phase via capillary movement.
• The stationary phase is polar inorganic solvent, i.e. water. Here, the absorbent paper supports the stationary phase, i.e. water.

Plant pigments are coloured organic substances derived from plants. Pigments absorb visible radiation between 380 nm (violet) and 760 nm (red).

They give colour to stems, leaves, flowers, and fruits. Also, they regulate processes like photosynthesis, growth, and development.

Plants produce various forms of pigments. Based on origin, function and water solubility, plant pigments are grouped into:

• Chlorophylls (green)
• Carotenoids (yellow, orange-red)
• Anthocyanins (red to blue, depending on pH)
• Betalains (red or yellow)

Chlorophyll : It is a green photosynthetic pigment. Chlorophyll a and b are present within the chloroplasts of plants. Because of the phytol side chain, they are water-repelling . Their structure resembles haemoglobin. But, they contain magnesium as a central metal instead of iron.

Carotenoids : These are yellow to yellow-orange coloured pigments. Also, they are very long water-repelling pigments. Carotenoids are present within the plastids or chromoplasts of plants.

Anthocyanins : These appear as red coloured pigments in vacuoles of plant cells. Anthocyanins are water-soluble pigments. They give pink-red colour to the petals, fruits and leaves.

Betalains : These are tyrosine derived water-soluble pigments in plants. Betacyanins (red-violet) and betaxanthins (yellow-orange) are the two pigments coming in this category. They are present in vacuoles of plant cells.

You can separate all the above pigments using paper chromatography.

Video: Separation of Plant Pigments

Preparation of Concentrated Leaf Extract

• Wash spinach leaves in distilled water.
• Then take out the spinach leaves and allow the moisture to dry out.
• After that, take a scissor and cut the leaves into the mortar.
• Take a little volume of acetone into the mortar. Note : Acetone is used instead of water to mash the leaves because it is less polar than the water. This allows a high resolution of the molecules in the sample between the absorbent paper.
• Then, grind spinach leaves using a pestle until liquid paste forms. Note : The liquid in the crushed leaf paste is the pigment extract.
• After that, take out the mixture into the watch glass or Petri dish.

Load the Leaf Extract onto Absorbent Paper

• Take Whatman filter paper and draw a line above 2 cm from the bottom margin. You can use a pencil and scale to draw a fainted line. Note : A pencil is used because pencil marks are insoluble in the solvent.
• Then, cut the filter paper to make a conical edge from the line drawn towards the margin end. You can use a scissor to cut the Whatman filter paper. Note : The conical end at the bottom of the filter paper results in better separation.
• Put a drop of leaf extract on the centre of a line drawn on the absorbent paper.
• Then, at the same time dry the absorbent paper.
• Repeat the above two steps many times so that the spot becomes concentrated enough.

Setup the Chromatography Chamber

• Take a clean measuring cylinder and add rising solvent (ether acetone) up to 4 ml.
• Bend the strip of paper from the top. Then, using a pushpin attach the paper to the bottom of the cork.
• Adjust the length of the paper. The absorbent paper should not touch the surface of the measuring cylinder.
• After that, allow the solvent to move up the absorbent paper.
• When the solvent front has stopped moving, remove the paper.
• Allow it to dry for a while until the colours completely elute from the paper.
• At last, mark the front edge travelled by each pigment.

Over the dried paper strip, you will see four different bands. Different colour streaks form because of different affinities with the mobile phase (solvent).

• The carotene pigment appears at the top as a yellow-orange band.
• A yellowish band appears below the carotene, which indicates xanthophyll pigment.
• Then a dark green band represents the chlorophyll-a pigment.
• The chlorophyll-b pigment appears at the bottom as a light green band.

Observation Table

1. Light green spot indicates chlorophyll-b pigment.

• Rf value= Distance chlorophyll-b travelled / Distance solvent travelled = 2/10 = 0.2

2. Dark green spot represents chlorophyll-a pigment.

• Rf value= Distance chlorophyll-a travelled / Distance solvent travelled = 3.7/10 = 0.37

3. The yellow band represents xanthophyll pigment.

• Rf value= Distance xanthophyll travelled / Distance solvent travelled = 5.6/10 = 0.56

4. The yellow-orange band indicates carotene pigment.

• Rf value= Distance carotene travelled / Distance solvent travelled = 9/10 = 0.9

Factors affecting the Rf values of a particular analyte are:

• Stationary phase
• The concentration of the stationary phase
• Mobile phase
• The concentration of the mobile phase
• Temperature

The Rf value of compounds in the mixture differs by any changes in the concentration of stationary and mobile phases.

Temperature affects the solvent capillary movement and the analyte’s solubility in the solvent. Rf value is independent of the sample concentration. Its value is always positive .

Related Topics:

• Difference Between Budding and Grafting
• Phototropism in Plants
• Potometer Experiment

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• Biology Article
• Separation Of Plant Pigments Through Paper Chromatography

How to Separate Plant Pigments Using Paper Chromatography

To distinguish and study the various pigments present in plants through the process of paper chromatography.

Plants carry out the process of photosynthesis, during which light energy from the sun is converted into chemical energy (food). The capturing of light energy is carried out by molecules known as pigments, which are present within the plant cells.

Explore more:   Plant cells .

What are Pigments?

Pigments are chemical compounds, which are able to reflect only a particular range of wavelengths of visible light. Leaves of plants primarily contain different types of pigments within their tissues. The four different types of pigments are listed below in a tabular column along with their colours.

In order to view and distinguish the primary four plant pigments, a simple technique known as chromatography can be used.

Read more:  Pigments

What is Chromatography?

It is a technique that is used to distinguish between different molecules. This differentiation is based on these attributes-shape, size, charge, mass, adsorption and solubility.

Types of chromatography:

• Column chromatography
• Paper chromatography
• Partition chromatography
• Thin-layer chromatography

Mechanism of Paper Chromatography

In this technique, the interaction between three components is involved – solid phase, separation of a mixture and a solvent.

• At first, the mixture is spotted onto the paper and is dried.
• The solvent is made to flow through the capillary attraction.
• While the solvent moves through the paper, the various components of the mixture differentiate into varied coloured spots.
• Later the paper is allowed to dry and the position of various compounds is viewed.
• The substance, which is the most soluble moves further on the paper as compared to the other substances that are less soluble.

Material Required

• Chromatography chamber
• Spinach leaves
• Mortar and pestle
• Ether acetone solvent
• Capillary tube
• Filter paper strips
• Watch glass
• In this experiment, spinach leaves are used to separate different pigments.
• Pick a few fresh and green leaves of spinach and wash it.
• Cut out small pieces of spinach using scissors. Add them to the mortar.
• Accurately measure 5ml acetone using a measuring cylinder and add it into the mortar.
• With the help of mortar and pestle, grind the spinach leaves into a smooth paste.
• Shift the prepared paste of spinach into the watch glass with the help of a spatula.
• Place a filter paper strip with a tapering notch towards one ending of the strip.
• Horizontally trace a line with a scale and a pencil that is 2 to 3 cm apart from the notch’s tip.
• Using a capillary tube, add 1 drop of the extract of the pigment in the midsection of the line.
• Let the drop dry. Repeat the same process of adding a drop and allowing it to dry for 4-5 times.
• In the chromatographic chamber, pour the ether acetone solvent.
• Make sure to folded and stapled an end side of the paper.
• Suspend the strip in the chamber.
• The loading spot remains about 1 cm above the level of the solvent.
• Let the chamber remain uninterrupted for a while.
• We can notice that the solvent passes along the paper scattering various pigments of the blend to different distances.
• Once the solvent reaches 3/4 th  of the strip, carefully take the strip off.
• Allow the strip to dry.

Observation

The dried paper strip displays four different bands. Discrete pigments can be distinguished with the help of colours.

• The Carotene pigment is observed at the topmost as an orange-yellow band of pigments distinctively.
• Just below this band, a yellowish band appears which indicates the pigment xanthophyll.
• The third band appearing dark green indicates chlorophyll-a pigment.
• The yellowish-green band present at the bottom is the chlorophyll b pigment.

Precautions

• The leaves that are selected should be green and fresh spinach leaves
• From the tip of the notch, the loading spot needs to be 2 to 3 cm apart
• While suspending the filter paper strips in the chamber, one need to ensure that the loading spot needs to be set up above 1 cm from the level of the solvent.

Viva Questions

Q.1. What Rf value or Retention factor?

A.1. The Retention factor or Rf value applies to chromatography to make the technique scientific. It is defined as the distance travelled by the compound divided by the distance travelled by the solvent.

Rf value = Distance travelled by the compound / Distance travelled by the solvent.

Q.2. What is Phycobilin?

A.2. Phycobilins are light-capturing bilins found in chloroplast organelles, cyanobacteria and in a few algae.

Q.3. What is the significance of pigment in photosynthesis?

A.3. It helps in the absorption of energy from light. The free electrons in the pigments present in their chemical structure transfer their energy to other molecules during photosynthesis when they turn into high energy electrons, thereby liberating energy they captured from light. This released energy is then used up by other molecules for the formation of sugars and related nutrients with the use of water and carbon dioxide.

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Shop Experiment Plant Pigment Chromatography Experiments​

Plant pigment chromatography.

Experiment #4A from Advanced Biology with Vernier

Introduction

Paper chromatography is a technique used to separate substances in a mixture based on the movement of the different substances up a piece of paper by capillary action. Pigments extracted from plant cells contain a variety of molecules, such as chlorophylls, beta carotene, and xanthophyll, that can be separated using paper chromatography. A small sample of plant pigment placed on chromatography paper travels up the paper due to capillary action. Beta carotene is carried the furthest because it is highly soluble in the solvent and because it forms no hydrogen bonds with the chromatography paper fibers. Xanthophyll contains oxygen and does not travel quite as far with the solvent because it is less soluble than beta carotene and forms some hydrogen bonds with the paper. Chlorophylls are bound more tightly to the paper than the other two, so they travel the shortest distance.

The ratio of the distance moved by a pigment to the distance moved by the solvent is a constant, R f . Each type of molecule has its own R f value.

In this experiment, you will

• Separate plant pigments.
• Calculate the R f values of the pigments.

Correlations

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This experiment is #4A of Advanced Biology with Vernier . The experiment in the book includes student instructions as well as instructor information for set up, helpful hints, and sample graphs and data.

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Paper Chromatography

How does paper chromatography work, stationary and mobile phases, paper chromatography experiment, applications.

Paper chromatography is a simple and cost-effective separation technique that separates and identifies different components in a mixture. [1-4]

In paper chromatography, a specialized paper acts as the stationary phase, while a liquid solvent is the mobile phase. The mixture to be analyzed is applied to the paper. As the solvent moves up through capillary action, it carries along the individual components of the mixture at different rates based on their solubility and affinity for the stationary phase.

The principle behind paper chromatography lies in the differential partitioning of compounds between the stationary and mobile phases. The stationary phase typically consists of cellulose fibers embedded in filter paper or thin-layer chromatography plates. These fibers provide an adsorbent surface for compounds to interact with.

Understanding the mechanism behind paper chromatography requires knowledge of several key processes. [1-4]

The first process is capillary action, which refers to the ability of liquids to flow through narrow spaces against gravity. In paper chromatography, capillary action allows the solvent to move up the paper strip due to its attraction to the fibers in the paper. As the solvent moves up, it carries the solutes in the analyzed mixture. This migration of solutes is driven by two main mechanisms: adsorption and partitioning.

Adsorption occurs when solute molecules adhere to the fibers or other surfaces within the paper. It can be influenced by polarity and molecular size, with more polar or larger molecules having stronger interactions with the stationary phase.

Conversely, partitioning involves solute molecules distributing themselves between two immiscible phases – in this case, between the stationary phase (paper) and mobile phase (solvent). The extent of partitioning depends on factors such as solute polarity and affinity for either phase.

As solutes migrate up through capillary action, they may experience different degrees of adsorption and partitioning along their journey. This results in their separation based on their characteristics. By analyzing how far each component migrates on a chromatogram – a visual representation of separated components – scientists can determine properties such as retention factor (R f ) values and identify unknown substances based on known reference compounds.

Stationary and mobile phases play crucial roles in separating components of a mixture. [1-4]

The stationary phase refers to the absorbent material fixed on the chromatography paper. It can be made of cellulose or other materials with high absorbency. The stationary phase acts as a substrate for the sample mixture to interact with during separation.

On the other hand, the mobile phase is the solvent or liquid that moves through the stationary phase, carrying the sample components. The mobile phase must have good solubility with the components of interest. It should be able to flow easily through the paper.

As the mobile phase moves through the stationary phase, it interacts differently with each mixture component based on their solubility and affinity for both phases. This differential interaction leads to separation as different components travel at different rates along the paper.

Choosing an appropriate combination of stationary and mobile phases is important for effective separation in paper chromatography. Factors such as polarity, viscosity, and compatibility between phases must be considered to achieve optimal results.

Performing a paper chromatography experiment involves several essential steps to ensure accurate results. The process begins with preparing samples for paper chromatography, then spotting the sample on the paper strip, and finally, developing the chromatogram. [1-4]

Preparing the samples is crucial in obtaining reliable data. It involves selecting appropriate substances to analyze and ensuring they are suitable for chromatography. Samples can be liquid or solid and must be dissolved or crushed into a solution before application.

Next, spotting the sample on the paper strip is done carefully to ensure accurate separation. A small spot of the prepared sample is placed near one end of a designated area on the filter paper strip. It is essential to use a capillary tube or micropipette for precise and consistent application.

Once all samples are spotted on the filter paper strip, it is time for the development of the chromatogram. This step involves placing one end of the strip into a solvent traveling up through capillary action. The choice of solvent depends on factors such as solubility and desired separation distance.

As the solvent moves up through the filter paper strip, it carries different components in each sample. These components separate based on their affinity for stationary (filter paper) and mobile (solvent) phases. The separation occurs due to differences in molecular size, polarity, or other physical properties.

Throughout this process, it is important to maintain controlled conditions such as temperature and humidity to ensure reproducibility. Further analysis can be conducted once an optimal separation has been achieved, which can take several minutes or hours depending on various factors, including solvent choice and sample composition.

The diverse applications of paper chromatography across various fields are listed below. [1-4]

• It plays a crucial role in forensic analysis by separating and identifying different components in complex mixtures, such as blood or ink samples.
• Aids in the analysis of crime scene evidence, allowing forensic scientists to determine the presence of specific substances and identify potential suspects based on chromatographic patterns
• Enables the separation of different dyes used in food coloring, helping to ensure compliance with regulatory standards and quality control measures
• Determines the authenticity and safety of food products by identifying and quantifying specific components present in complex food matrices
• Separate and identify active ingredients, impurities, and by-products in pharmaceutical formulations.
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The below mentioned article includes a list of three experiments on paper chromatography.

1. Experiment to separate amino acids in a mixture by paper strip chromatography:

It is based on the fact that paper chromato­graphy separates compounds on the basis of their different rates of migration of filter paper (cellulose). The rates of migration depend upon the solvent which is flowing up or down the paper and on the relative adsorption which holds the molecules more or less tightly to the paper.

If the solvent flows towards upper side on the paper it is called ascending chromatogram, and if it flows towards lower side then it is known as descending chromatogram.

Requirements:

Chromatography paper, test tubes (2), capillary tubes (3), coupling jar (1), distilled water, amino acids (glycine and aspartic acid), phenol and ninhydrin.

1. Take three stips of chromatography paper of equal size (12 cm in length and 1.5 cm in width).

2. Draw a fine line with a lead pencil, parallel to and 1.5 cm from one edge of the paper. This line will indicate the bottom of your chromatogram.

3. On this line draw a circle on each strip, about 1.5 cm from one edge. These circles will indicate the position of your samples.

4. Put a drop of glycine in the circle of one strip, aspartic acid on the second and that of a mixture of both amino acids on the 3rd strip with the help of separate capillary tubes. (Note : Avoid excess handling of the chromatography paper, since your hands may contaminate it with amino acids. Touch it only at the edges).

5. Take 5 ml of 80% phenol in three large test tubes.

6. Fix each strip in the cork as shown in Fig. 40.

7. Insert one strip in each test tube and see carefully that lower edge of the strip touches the phenol. (Note carefully that the amino acid spot should not he touched by the phenol and the paper should not touch the wall of the test tubes).

8. Keep these test tubes in the stand and wait for 20 to 30 minutes or more until the solvent has risen within 0.5 cm from the top of the paper.

9. Remove the strips from the tube and let it dry in an oven at about 100°C for 3-5 minutes.

10. Dry strips should be sprayed by 0.1 % ninhydrin- acetone reagent and set it aside to dry.

11. Keep it again in the oven for 2 to 3 minutes. Do not overheat the paper.

12. Remove the paper and immediately outline with pencil the spots that you see. Amino acids will appear as purple spots on the filter paper.

Rf value can be measured by the following formula:

5. About 2 cm of the jar is filled with the solvent (petroleum ether: acetone = 100:12).

6. Insert the strip attached with the hook in the jar and see carefully that lower edge of the strip touches the solvent. Note carefully that chlorophyll spot should not be touched by the solvent and paper should not touch the wall of the jar.

7. Keep the whole apparatus as such for 20 to 30 minutes and see that solvent has risen nearly up to the top of the paper.

Different pigments separate at different levels on filter paper strip.

In this experiment different pigments from below upward separate in a sequence of chlorophyll b, chlorophyll a, xanthophyll’s and carotene.

Separation of different pigments on strip is based on the fact that paper chromatography separates compounds on the basis of their different rates of migration on filter paper (cellulose). The rate of migration depends upon the solvent which is flowing up and also on the relative adsorption which holds the molecules more or less tightly to the paper.

Related Articles:

• Types of Chromatography (With Diagram) | Proteins | Biology
• Separation of Amino Acids by Paper Chromatography (With Diagram)

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Paper Chromatography Report

Introduction The purpose of this experiment is to observe how chromatography can be used to separate mixtures of chemical substances. Chromatography serves mainly as a tool for the examination and separation of mixtures of chemical substances. Chromatography is using a flow of solvent or gas to cause the components of a mixture to migrate differently from a narrow starting point in a specific medium, in the case of this experiment, filter paper. It is used for the purification and isolation of various substances. A chromatographically pure substance is the result of the separation. Because purification of substances is required to determine their properties, chromatography is an indispensable tool in the sciences concerned with chemical substances and their reactions.

Chromatography is also used to compare and describe chemical substances. The chromatographic sequence of sorbed substances is related to their atomic and molecular structures. A change in a chemical substance produced by a chemical or biological reaction often alters the solubility and migration rate. With this knowledge, alterations or changes can be detected in the substance.

In all chromatographic separations, there is an important relationship between the solvent, the chromatography paper, and the mixture. For a particular mixture, the solvent and the paper must be chosen so the solubility is reversible and be selective for the components of the mixture. The main requirement, though, of the solvent is to dissolve the mixture needing to be separated. The porous paper used  must also absorb the components of the mixtures selectively and reversibly. For the separation of a mixture, the substances making up the mixture must be evenly dispersed in a solution, a vapor, or a gas. Once all of the above criteria have been met, chromatography can be a simple tool for separating and comparing chemical mixtures.

Hypothesis Paper can be used to separate mixed chemicals.

Materials The materials used for this lab are paper, pencil, eraser, filter paper, test tube, rubber stopper, paper clip, metric ruler, black felt-tip pen, and a computer.

Methods The first step of the method is to bend a paper clip so that it is straight with a hook at one end. Push the straight end of the paper clip into the bottom of the rubber stopper. Next, you hang a thin strip of filter paper on the hooked end of the paper clip. Insert the paper strip into the test tube. The paper should not touch the sides of the test tube and should almost touch the bottom of the test tube. Now you will remove the paper strip from the test tube. Draw a solid 5-mm-wide band about 25 mm from the bottom of the paper, using the black felt-tip pen. Use a pencil to draw a line across the paper strip 10 cm above the black band.

Pour about 2 mL of water into the test tube. The water will act as a solvent. Put the filter paper back into the test tube with the bottom of the paper in the water and the black band above the water. Observe what happens as the liquid travels up the paper. Record the changes you see. When the solvent has reached the pencil line, remove the paper from the test tube. Measure how far the solvent traveled before the strip dries. Finally, let the strip dry on the desk. With the metric ruler, measure the distance from the starting point to the top edge of each color. Record this data in a data table. Calculate a ratio for each color by dividing the distance the color traveled by the distance the solvent traveled.

Results The results of the experiment are shown in a chart and a graph.

1. How many colors separated from the black ink? Five colors separated from the black ink: yellow, pink, red, purple, and blue.

2. What served as the solvent for the ink? Water served as the solvent for the ink. As the solvent traveled up the paper, which color of ink appeared first? The color orange first appeared as the solvent traveled up the paper.

3. List the colors in order, from top to bottom, which separated from the black ink. The colors separated in this order, from top to bottom: blue, purple, red, pink, and then yellow.

4. In millimeters, how far did the solvent travel? The solvent traveled 111 mm.

5. From your results, what can you conclude is true about black ink? Black ink is a mixture of several different colors.

6 . Why did the inks separate? The inks separated because the black ink was a mixture of different pigments with different molecular characteristics. These differences allow for different rates of absorption by the filter paper.

7. Why did some inks move a greater distance? The ink least readily absorbed by the paper would then travel the farthest from the starting mark. You can conclude from this information that the different pigments were absorbed at different rates.

Error Analysis Possible errors could include inaccurate measurements of the distances traveled by the inks and mistakes when calculating the ratio traveled by the water and colors. If a longer test tube was used, a longer strip of filter paper could have been used. This may have changed the ratios. Another color may have been present, but not detected because of the filter paper length.

Conclusion The proposed hypothesis was correct. The paper chromatography did show that black ink could be separated into various colors. The black ink gets its color from a mixture of various colored inks blended together. The first color of ink to appear on the filter paper was yellow followed by pink, red, purple then blue. The colors separated the way they did because of the differences in their molecular characteristics, specifically, their solubility in water and their rate of absorption by the paper. The most soluble and readily absorbed ink color was the yellow. The least soluble and least absorbable ink color was the blue.

Microbe Notes

Paper Chromatography- Definition, Types, Principle, Steps, Uses

Table of Contents

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What is Paper Chromatography?

Paper chromatography (PC) is a type of planar chromatography whereby chromatography procedures are run on a specialized paper.

PC is considered to be the simplest and most widely used of the chromatographic techniques because of its applicability to isolation, identification, and quantitative determination of organic and inorganic compounds.

It was first introduced by German scientist Christian Friedrich Schonbein (1865).

Types of Paper chromatography

Paper adsorption chromatography.

Paper impregnated with silica or alumina acts as adsorbent (stationary phase) and solvent as mobile phase.

Paper Partition Chromatography

Moisture / Water present in the pores of cellulose fibers present in filter paper acts as stationary phase & another mobile phase is used as solvent In general paper chromatography mostly refers to paper partition chromatography. 

Principle of Paper chromatography

The principle of separation is mainly partition rather than adsorption. Substances are distributed between a stationary phase and a mobile phase. Cellulose layers in filter paper contain moisture which acts as a stationary phase. Organic solvents/buffers are used as mobile phase. The developing solution travels up the stationary phase carrying the sample with it. Components of the sample will separate readily according to how strongly they adsorb onto the stationary phase versus how readily they dissolve in the mobile phase.

Instrumentation of Paper chromatography

• Stationary phase & papers used
• Mobile phase
• Developing Chamber
• Detecting or Visualizing agents

1. STATIONARY PHASE AND PAPERS

• Whatman filter papers of different grades like No.1, No.2, No.3, No.4, No.20, No.40, No.42 etc
• In general the paper contains 98-99% of α-cellulose, 0.3 – 1% β -cellulose.

Other modified papers

• Acid or base washed filter paper
• Glass fiber type paper.
• Hydrophilic Papers – Papers modified with methanol, formamide, glycol, glycerol etc.
• Hydrophobic papers – acetylation of OH groups leads to hydrophobic nature, hence can be used for reverse phase chromatography.
• Impregnation of silica, alumna, or ion exchange resins can also be made.

2. PAPER CHROMATOGRAPHY MOBILE PHASE

• Pure solvents, buffer solutions or mixture of solvents can be used.

Hydrophilic mobile phase

• Isopropanol: ammonia:water 9:1:2
• Methanol : water 4:1
• N-butanol : glacial acetic acid : water 4:1:5

Hydrophobic mobile phases

• dimethyl ether: cyclohexane kerosene : 70% isopropanol
• The commonly employed solvents are the polar solvents, but the choice depends on the nature of the substance to be separated.
• If pure solvents do not give satisfactory separation, a mixture of solvents of suitable polarity may be applied.

3. CHROMATOGRAPHIC CHAMBER

• The chromatographic chambers are made up of many materials like glass, plastic or stainless steel . Glass tanks are preferred most.
• They are available invarious dimensional size depending upon paper length and development type.
• The chamber atmosphere should be saturated with solvent vapor.

Steps in Paper Chromatography

In paper chromatography, the sample mixture is applied to a piece of filter paper, the edge of the paper is immersed in a solvent, and the solvent moves up the paper by capillary action. The basic steps include:

Selection of Solid Support

Fine quality cellulose paper with defined porosity, high resolution, negligible diffusion of the sample, and favoring good rate of movement of solvent.

Selection of Mobile Phase

Different combinations of organic and inorganic solvents may be used depending on the analyte.

Example. Butanol: Acetic acid: Water (12:3:5) is a suitable solvent for separating amino acids.

Saturation of Tank

The inner wall of the tank is wrapped with filter paper before the solvent is placed in the tank to achieve better resolution.

Sample Preparation and Loading

If the solid sample is used, it is dissolved in a suitable solvent. Sample (2-20ul) is added on the baseline as a spot using a micropipette and air dried to prevent the diffusion.

Development of the Chromatogram

Different types of development techniques can be used:

ASCENDING DEVELOPMENT

• Like conventional type, the solvent flows against gravity.
• The spots are kept at the bottom portion of paper and kept in a chamber with mobile phase solvent at the bottom.

DESCENDING TYPE

• This is carried out in a special chamber where the solvent holder is at the top.
• The spot is kept at the top and the solvent flows down the paper.
• In this method solvent moves from top to bottom so it is called descending chromatography.

ASCENDING – DESCENDING DEVELOPMENT

• A hybrid of above two techniques is called ascending-descending chromatography.
• Only length of separation increased, first ascending takes place followed by descending.

CIRCULAR / RADIAL DEVELOPMENT

• Spot is kept at the centre of a circular paper.
• The solvent flows through a wick at the centre & spreads in all directions uniformly.

Drying of Chromatogram

After the development, the solvent front is marked and left to dry in a dry cabinet or oven.

Colorless analytes were detected by staining with reagents such as iodine vapor, ninhydrin, etc.

Radiolabeled and fluorescently labeled analytes were detected by measuring radioactivity and fluorescence respectively.

Some compounds in a mixture travel almost as far as the solvent does; some stay much closer to the baseline . The distance traveled relative to the solvent is a constant for a particular compound as long as other parameters such as the type of paper and the exact composition of the solvent are constant. The distance traveled relative to the solvent is called the Rf value.

Thus, in order to obtain a measure of the extent of movement of a component in a paper chromatography experiment, “Rf value” is calculated for each separated component in the developed chromatogram. An Rf value is a number that is defined as the distance traveled by the component from the application point.

Applications of Paper Chromatography

• To check the control of purity of pharmaceuticals,
• For detection of adulterants,
• Detect the contaminants in foods and drinks,
• In the study of ripening and fermentation,
• For the detection of drugs and dopes in animals & humans
• In analysis of cosmetics
• Analysis of the reaction mixtures in biochemical labs.

Advantages of Paper Chromatography

• Paper Chromatography requires very less quantitative material.
• Paper Chromatography is cheaper compared to other chromatography methods.
• Both unknown inorganic as well as organic compounds can be identified by paper chromatography method.
• Paper chromatography does not occupy much space compared to other analytical methods or equipments.
• Excellent resolving power

Limitations of Paper Chromatography

• Large quantity of sample cannot be applied on paper chromatography.
• In quantitative analysis paper chromatography is not effective.
• Complex mixture cannot be separated by paper chromatography.
• Less Accurate compared to HPLC or HPTLC
• http://frndzzz.com/Advantages-and-Disadvantages-of-Paper-Chromatography
• https://www.slideshare.net/shaisejacob/paper-chromatography-pptnew?next_slideshow=1
• https://www.slideshare.net/shaisejacob/paper-chromatography-ppt-new
• https://www.biochemden.com/paper-chromatography/
• http://web.engr.oregonstate.edu/~rochefow/K-12%20Outreach%20Activities/Microfluidics%20&%20Pregnancy%20Test%20Kit%20Lab/paper%20chromatography_Chemguide.pdf
• https://pubs.acs.org/doi/abs/10.1021/ac60051a002

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Sagar Aryal

6 thoughts on “Paper Chromatography- Definition, Types, Principle, Steps, Uses”

Examples of substances that can be effectively separated by paper chromatography is necessary.

I enjoy this write up, but the definition of Rf Values just mention distance traveled by the solute from the point from the point of application of the sample, how about the total distance traveled by the solvent? Some examples of how Rf value can be calculated is necessary.

Pls how is charging a disadvantage of filter paper in chromatography

Hi! I enjoyed reading it. Hmm just wanna know somepoints.

Why it is best to use the farthest distance traveled by a sugar if and when the solvent went over the paper and what is the purpose of heating the chromatographic paper after running the procedure?

This links are so much useful and it’s very helpful also….

Thanks for sharing important details like this. I enjoyed reading your site, and love to know the latest updates.

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Paper Chromatography

• Solubility:  If the components of the mixture are soluble in the solvent being used, the mixture will be carried up the paper strip as the solvent travels.  If the material is soluble, the mixture will dissolve as the solvent front moves through it.  If the material is a mixture of substances, some of these substances will likely be more or less soluble than others.  The more soluble substances will move faster and to a greater distance than those that are less soluble.
• Molecular Weight:  Those substances of lighter molecular weight will move higher up the paper than those substances having a higher molecular weight.
• The chromatography paper is made of cellulose, a polar substance, and the compounds within the mixture travel farther if they are non-polar. More polar substances bond with the cellulose paper more quickly, and therefore do not travel as far.

• Chromatography paper
• Some sort of container (beaker or test tube) 
• A mixture that can be separated

7 comments:

Hi, why is paper chromatography used in schools and not over methods(TLC,Gas,Column)? Thank you for your help.

As it is simpler and very visual! It's a good precursor to TLC, Gas, Column at a higher level :)

I'm doing paper chromatography for a science fair project, thanks for the help!! (Going on my bibliography)

This is really, really nice!

What leaf pigment was used in this experiment?

What specific type of leaf pigment was used in this experiment?

I most often buy a bag of fresh spinach at the grocery store. It is easy to grind up and always gives great results. I have also pulled leaves from bushes around my school and that worked, too!

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paper chromatography

Our editors will review what you’ve submitted and determine whether to revise the article.

• Open Library Publishing Platform - DRAFT – Organic and Biochemistry Supplement to Enhanced Introductory College Chemistry - Thin Layer (TLC) And Paper Chromatography (PC)
• Chemistry LibreTexts Library - Paper Chromatography - Separation and Identification of Five Metal Cations
• Oregon State University - College of Engineering - Paper chromatography
• Academia - Paper Chromatography
• Journal of Emerging Technologies and Innovative Research - Paper Chromatography: A Review

paper chromatography , in analytical chemistry , technique for separating dissolved chemical substances by taking advantage of their different rates of migration across sheets of paper. It is an inexpensive but powerful analytical tool that requires very small quantities of material.

The method consists of applying the test solution or sample as a spot near one corner of a sheet of filter paper. The paper is initially impregnated with some suitable solvent to create a stationary liquid phase . An edge of the paper close to the test spot is then immersed in another solvent in which the components of the mixture are soluble in varying degrees. The solvent penetrates the paper by capillary action and, in passing over the sample spot, carries along with it the various components of the sample. The components move with the flowing solvent at velocities that are dependent on their solubilities in the stationary and flowing solvents. Separation of the components is brought about if there are differences in their relative solubilities in the two solvents. Before the flowing solvent reaches the farther edge of the paper, both solvents are evaporated , and the location of the separated components is identified, usually by application of reagents that form coloured compounds with the separated substances. The separated components appear as individual spots on the path of the solvent. If the solvent flowing in one direction is not able to separate all the components satisfactorily, the paper may be turned 90° and the process repeated using another solvent.

Paper chromatography has become standard practice for the separation of complex mixtures of amino acids , peptides , carbohydrates , steroids , purines , and a long list of simple organic compounds . Inorganic ions can also readily be separated on paper. Compare thin-layer chromatography .

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Behaviour and peculiarities of oil hydrocarbon removal from rain garden structures.

1. Introduction

• Determine the effectiveness of rain garden designs in removing petroleum hydrocarbons, such as diesel fuel and used engine oil, from rainwater;
• Study the temporal and spatial characteristics of the distribution and accumulation of petroleum hydrocarbons in experimental rain gardens;
• Study the impact of petroleum hydrocarbons on plants of the species Physocarpus opulifolia Diabolo ;
• Provide recommendations for the implementation and maintenance of rain gardens in real environmental conditions, in particular in areas with high levels of oil pollution, such as petrol stations, car parks, and car washes.

2. Materials and Methods

2.1. experimental filter columns (cylinders), 2.2. characteristics of experimental samples, 2.3. calculation of model solution volumes, 2.4. reverse-phase high-performance liquid chromatography, 2.5. experimental procedure with filter columns, 2.6. study of plant resistance, 3. results and discussion, 3.1. analysis of the leachate from the experimental columns, 3.2. analysis of soil media, 3.3. changes in hydraulic conductivity, 3.4. plant resistance, 4. conclusions, 5. recommendations.

• To increase the service life of rain gardens designed to filter stormwater from oil products, it is recommended to supplement the bioretention soil medium with highly effective adsorbents to optimise the removal of pollutants, such as activated carbon, biochar, and ash. This is in line with the results of [ 77 ], which indicate that soil supplemented with activated carbon can extend the life of a rain garden by approximately 10–20 years before the environment is completely saturated with pollutants.
• When designing rain gardens, it is necessary to take into account the type of vegetation and the size of the systems, which have important impacts on hydraulic conductivity and the degree of efficiency of stormwater treatment. Plant roots play an important role in the formation of macropores in the media, which help to maintain infiltration rates, aerate the soil, and minimise compaction. The desorption of tightly bound petroleum hydrocarbons from soil matrices is achieved through the interaction between oil and biologically active compounds released by plants and microorganisms. In addition, plant root microbes can break down aggregates and freely bind hydrocarbons from nanopores. The most effective plant species are those with a developed fibrous root system and thick roots. However, the presence of vegetation significantly complicates the regular maintenance and cleaning of the system surface. An appropriate solution may be to divide the system into zones with vegetation and zones without vegetation (with different maintenance intervals). It should also be noted that the vegetation will need to be replaced after about ten years due to the loss of the decorative effect associated with age-related changes [ 92 ].
• Petroleum hydrocarbons accumulate in the surface layers of the soil medium of rain garden structures, and the concentrations of pollutants in the media decrease with depth. Thus, the design of a rain garden aimed at reducing the concentration of petroleum hydrocarbons can be shallow, and the process of reducing the concentration should be focused on the upper surface layer near the inlet, where the maximum accumulation of pollutants occurs. It is recommended to periodically replace the surface layer of the rain garden structure by removing and replacing the fill materials, which will maintain the effectiveness of the system over time. Although there are guidelines for maintaining the effectiveness of rain gardens [ 93 ], they do not provide specific timelines for replacing soil media. Therefore, it is recommended that the topsoil be replaced when the structure is overflowing or saturated with water for more than 48 h after a rain event. This means that the concentration of contaminants in the system layer has reached values that disrupt hydrological processes in the structure. This recommendation is consistent with the results of a study conducted in North Carolina by researchers [ 94 ]. They repaired two rain garden systems by excavating the top substrate to a thickness of 7.5 cm. This resulted in an increase in infiltration rates of up to 10 times and a 90% increase in the water storage surface area. The volume of overflow in the two systems, which was 37% and 35%, respectively, in the pre-repair condition, decreased to 12% and 11%, respectively, after the repair.

Author Contributions

Institutional review board statement, informed consent statement, data availability statement, conflicts of interest.

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• SINTEF Open: Review of Stormwater Management Practices. Available online: https://sintef.brage.unit.no/sintef-xmlui/handle/11250/2447971 (accessed on 26 March 2024).
• Zhang, J.; Li, R.; Zhang, X.; Ding, C.; Hua, P. Traffic contribution to polycyclic aromatic hydrocarbons in road dust: A source apportionment analysis under different antecedent dry-weather periods. Sci. Total Environ. 2019 , 658 , 996–1005. [ Google Scholar ] [ CrossRef ]
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Share and Cite

Kravchenko, M.; Trach, Y.; Trach, R.; Tkachenko, T.; Mileikovskyi, V. Behaviour and Peculiarities of Oil Hydrocarbon Removal from Rain Garden Structures. Water 2024 , 16 , 1802. https://doi.org/10.3390/w16131802

Kravchenko M, Trach Y, Trach R, Tkachenko T, Mileikovskyi V. Behaviour and Peculiarities of Oil Hydrocarbon Removal from Rain Garden Structures. Water . 2024; 16(13):1802. https://doi.org/10.3390/w16131802

Kravchenko, Maryna, Yuliia Trach, Roman Trach, Tetiana Tkachenko, and Viktor Mileikovskyi. 2024. "Behaviour and Peculiarities of Oil Hydrocarbon Removal from Rain Garden Structures" Water 16, no. 13: 1802. https://doi.org/10.3390/w16131802

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