separation of sugars by paper chromatography experiment pdf

Analytical Chemistry

Paper Chromatography

What is paper chromatography.

Chromatography technique that uses paper sheets or strips as the adsorbent being the stationary phase through which a solution is made to pass is called paper chromatography. It is an inexpensive method of separating dissolved chemical substances by their different migration rates across the sheets of paper. It is a powerful analytical tool that uses very small quantities of material. Paper chromatography was discovered by Synge and Martin in the year 1943.

Paper chromatography principle, paper chromatography diagram, paper chromatography procedure, paper chromatography applications.

Types of Paper Chromatography
Frequently Asked Questions – FAQs

The principle involved can be partition chromatography or adsorption chromatography. Partition chromatography because the substances are partitioned or distributed between liquid phases. The two phases are water held in pores of the filter paper and the other phase is a mobile phase which passes through the paper. When the mobile phase moves, the separation of the mixture takes place. The compounds in the mixture separate themselves based on the differences in their affinity towards stationary and mobile phase solvents under the capillary action of pores in the paper. Adsorption chromatography between solid and liquid phases, wherein the solid surface of the paper is the stationary phase and the liquid phase is the mobile phase.

Below we have explained the procedure to conduct Paper Chromatography Experiment for easy understanding of students.

Selecting a suitable type of development: It is decided based on the complexity of the solvent, paper, mixture, etc. Usually ascending type or radial paper chromatography is used as they are easy to perform. Also, it is easy to handle, the chromatogram obtained is faster and the process is less time-consuming.
Selecting a suitable filter paper : Selection of filter paper is done based on the size of the pores and the sample quality.
Prepare the sample: Sample preparation includes the dissolution of the sample in a suitable solvent (inert with the sample under analysis) used in making the mobile phase.
Spot the sample on the paper: Samples should be spotted at a proper position on the paper by using a capillary tube.
Chromatogram development: Chromatogram development is spotted by immersing the paper in the mobile phase. Due to the capillary action of paper, the mobile phase moves over the sample on the paper.
Paper drying and compound detection : Once the chromatogram is developed, the paper is dried using an air drier. Also, detecting solution can be sprayed on the chromatogram developed paper and dried to identify the sample chromatogram spots.

There are various applications of paper chromatography . Some of the uses of Paper Chromatography in different fields are discussed below:

To study the process of fermentation and ripening.
To check the purity of pharmaceuticals.
To inspect cosmetics.
To detect the adulterants.
To detect the contaminants in drinks and foods.
To examine the reaction mixtures in biochemical laboratories.
To determine dopes and drugs in humans and animals.

Types of paper chromatography:

Ascending Paper Chromatography – The techniques goes with its name as the solvent moves in an upward direction.
Descending Paper Chromatography – The movement of the flow of solvent due to gravitational pull and capillary action is downwards, hence the name descending paper chromatography.
Ascending – Descending Paper Chromatography – In this version of paper chromatography, movement of solvent occurs in two directions after a particular point. Initially, the solvent travels upwards on the paper which is folded over a rod and after crossing the rod it continues with its travel in the downward direction.
Radial or Circular Paper Chromatography – The sample is deposited at the centre of the circular filter paper. Once the spot is dried, the filter paper is tied horizontally on a Petri dish which contains the solvent.
Two Dimensional Paper Chromatography – Substances which have the same r f values can be resolved with the help of two-dimensional paper chromatography.

Frequently Asked Questions – FAQs

What are the advantages of paper chromatography.

Paper Chromatography Has Many Benefits Simple and rapid Paper chromatography necessitates a minimal amount of quantitative material. Paper chromatography is less expensive than other chromatography methods. The paper chromatography method can identify both unknown inorganic and organic compounds. Paper chromatography takes up little space when compared to other analytical methods or equipment. Outstanding resolving power

Why water is not used in paper chromatography?

It is preferable to use a less polar solvent, such as ethanol, so that the non-polar compounds will travel up the paper while the polar compounds will stick to the paper, separating them.

What are the limitations of Paper Chromatography?

Limitations of Paper Chromatography are as follows- Paper chromatography cannot handle large amounts of sample. Paper chromatography is ineffective in quantitative analysis. Paper chromatography cannot separate complex mixtures. Less Accurate than HPLC or HPTLC

What is the importance of paper chromatography?

Paper chromatography has traditionally been used to analyse food colours in ice creams, sweets, drinks and beverages, jams and jellies. Only edible colours are permitted for use to ensure that no non-permitted colouring agents are added to the foods. This is where quantification and identification come into play.

Is paper chromatography partition or adsorption?

A type of partition chromatography is paper chromatography.

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Published: 23 September 1950

Separation and Identification of Sugars using Paper Chromatography

L. BOGGS 1 ,
L. S. CUENDET 1 ,
I. EHRENTHAL 1 ,
R. KOCH 1 &
F. SMITH 1

Nature volume 166 , pages 520–521 ( 1950 ) Cite this article

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CHROMATOGRAPHY on filter paper has provided an excellent and a much-needed method for the investigation of the structure of polysaccharides 1–5 and proteins 6–10 . While butanol–ethanol–water, butanol–acetic acid–water, and phenol–water are effective mixtures, among others 1 , for the separation of a wide variety of sugars 11,12,5 and their methyl derivatives 13 , the phenol–water mixture is usually better than either of the other two mentioned above for the separation of unmethylated sugars (cf. ref. 14). The method using phenol–water, however, suffers from the disadvantage that the location of the sugars on the paper by treatment with ammoniacal silver nitrate cannot be accomplished without some difficulty because the paper quickly becomes dark or sometimes black. It is claimed that suitable purification of the phenol either reduces the darkening 5 or eliminates it altogether 15 . We find that most of the darkening can be avoided by extracting the paper with ether before spraying with the silver reagent. By adopting this slight modification, it has been found unnecessary to purify the phenol.

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BOGGS, L., CUENDET, L., EHRENTHAL, I. et al. Separation and Identification of Sugars using Paper Chromatography. Nature 166 , 520–521 (1950). https://doi.org/10.1038/166520b0

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Two-step macromolecule separation process with acid pretreatment and high-shear-assisted extraction for microalgae-based biorefinery, 1. introduction, 2. materials and methods, 2.1. materials, 2.2. biochemical composition analysis, 2.3. acid pretreatment of wet biomass, 2.4. high shear-assisted lipid extraction, 2.5. characterization of oil, water, and solid phase products, 3. results and discussion, 3.1. biochemical composition of chlorella sp. abc-001, 3.2. acid pretreatment of concentrated wet biomass, 3.3. effects of mixing efficiency on the lipid recovery process, 3.4. separation of macromolecules in post-extracted mixture: biorefinery concept, 3.4.1. lipid phase: esterifiable lipids recovery, 3.4.2. water phase: fermentable glucose recovery, 3.4.3. solid phase: maintaining protein integrity, 3.4.4. impurities removal effects on extracted lipids, 3.5. comparison with other biorefinery processes, 4. conclusions, author contributions, institutional review board statement, informed consent statement, data availability statement, conflicts of interest.

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Click here to enlarge figure

Composition		Biomass (wt.%)	Crude Lipids (wt.%)	CASS Process
Composition		Biomass (wt.%)	Crude Lipids (wt.%)	Oil Phase (wt.%)	Water Phase (wt.%)	Solid Phase (wt.%)
Macromolecule composition	Lipid	35.7 ± 0.6	35.7 ± 0.6	30.1 ± 0.2	N/A	2.2 ± 0.01
	Esterifiable Lipid (FAME)	29.8 ± 0.2	29.8 ± 0.2	27.0 ± 0.5	N/A	0.5 ± 0.2
	Carbohydrate	40.1 ± 1.3	N/A	N/A	29.1 ± 0.8	2.9 ± 0.1
	Glucose	19.1 ± 0.1			17.2 ± 0.3	0.2 ± 0.1
	Protein	18.7 ± 2.3			N/A	19.0 ± 0.4
	Ash	5.5 ± 0.1	0.1 ± 0.1	<0.1	4.9 ± 0.0	0.5 ± 0.0
Elemental composition	C	51.8 ± 0.1	74.1 ± 0.6	75.4 ± 0.4	N/A	44.5 ± 0.1
	H	8.2 ± 0.1	11.8 ± 0.1	12.2 ± 0.1		6.5 ± 0.1
	O	29.7 ± 0.1	11.8 ± 0.1	11.4 ± 0.1		27.4 ± 0.0
	N	2.6 ± 0.0	0.4 ± 0.1	0.2 ± 0.0		8.2 ± 0.1
	S	0.2 ± 0.1	N/D	N/D		2.5 ± 0.1

Content	Biomass (wt. ppm)	Crude Lipid (wt. ppm)	CASS Process
Content	Biomass (wt. ppm)	Crude Lipid (wt. ppm)	Oil Phase (wt. ppm)	Water Phase (wt. ppm)	Solid Phase (wt. ppm)
P	27,021 ± 49	1130 ± 56	206.5 ± 1.9	24,318 ± 155	3346 ± 155
Ca	3168 ± 69	74.5 ± 5.1	N/D	2738 ± 21	461.2 ± 5.3
Na	623 ± 5	N/D	N/D	792.8 ± 6.7	137.4 ± 3.3
K	19,844 ± 509	65.3 ± 1.8	N/D	17,344 ± 242	312.9 ± 2.0
Cu	889 ± 37	271.8 ± 8.2	24.9 ± 1.3	700.7 ± 0.4	352.1 ± 5.1
Zn	232 ± 53	17.4 ± 1.8	N/D	298.1 ± 0.7	N/D
Mg	3325 ± 44	N/D	N/D	2891 ± 5	574.0 ± 10.2

Process	Strains	Biomass Conc. (g/L)	Operating Conditions	Macromolecule Yield (%)	References
High-pressure homogenizer	Nannochloropsis sp.	110–230	(1) Cell weakening (35 °C, 7–24 h) (2) High-pressure homogenizer (800–1000 bar, 1 pass) (3) Lipid recovery (2 h, biomass (paste):solvent 5:2 w/w)	Lipid: 25 Carbohydrate in water: 41 Protein in biomass: 51	[ ]
Acid hydrolysis	Tribonema sp.	50	(1) Acid hydrolysis (3 wt%, sulfuric acid, 121 °C, 45 min) (2) Lipid recovery (ethanol-hexane (1:3, v/v) on shaker incubation, 2.5 h, 50 °C, biomass (paste):solvent = 1:6 w/v)	Biodiesel: 98.5 Monosaccharides: 81.5	[ ]
Acid hydrolysis	Chlorella sp.	250	(1) Acid hydrolysis (2 wt%, sulfuric acid, 155 °C, 10 min) (2) Lipid recovery (hexane, 2 h, biomass (paste):solvent 1:1 v/v)	Lipid: 22.2 Glucose: 81.2	[ ]
Acid hydrolysis	Scenedesmus sp.	250		Lipid: 92.5 Glucose: 73.1	[ ]
Acid hydrolysis	Chlorella sp. ABC-001	50	(1) Acid hydrolysis (0.1 N, sulfuric acid, 170 °C, 4 min) (2) Phase separation using centrifugation (1000 rpm, 5 min) (3) Lipid recovery (hexane with vortexing, 20 min)	Lipid: 100 Monosaccharides: 89	[ ]
Hydrothermal liquefaction	Chlorella sp. KR1	50	(1) Hydrothermal liquefaction (180 °C, 1 h) (2) Lipid recovery (hexane with sonication, 2 h)	Lipid: 87 Glucose: 70	[ ]
CASS	Chlorella sp. ABC-001	50	(1) Acid hydrolysis (5 wt.% sulfuric acid, 100 °C, 1 h) (2) Lipid recovery (hexane with high-shear mixing, 3000 rpm, 30 min, biomass (wet):solvent = 1:1 v/v)	Lipid: 90.6 Glucose: 90.0 Protein: 100	This study

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Kim, D.; Kang, S.-G.; Chang, Y.K.; Kwak, M. Two-Step Macromolecule Separation Process with Acid Pretreatment and High-Shear-Assisted Extraction for Microalgae-Based Biorefinery. Sustainability 2024 , 16 , 7589. https://doi.org/10.3390/su16177589

Kim D, Kang S-G, Chang YK, Kwak M. Two-Step Macromolecule Separation Process with Acid Pretreatment and High-Shear-Assisted Extraction for Microalgae-Based Biorefinery. Sustainability . 2024; 16(17):7589. https://doi.org/10.3390/su16177589

Kim, Donghyun, Seul-Gi Kang, Yong Keun Chang, and Minsoo Kwak. 2024. "Two-Step Macromolecule Separation Process with Acid Pretreatment and High-Shear-Assisted Extraction for Microalgae-Based Biorefinery" Sustainability 16, no. 17: 7589. https://doi.org/10.3390/su16177589

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To study Separation of sugar by Paper chromatography
To study Separation of sugar by Paper chromatography
Paper chromatography
What Is Paper Chromatography and How Does it Work?
Paper Chromatography- Definition, Types, Principle, Steps, Uses
Separation of Sugars Using Paper Chromatography

VIDEO

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PAPER CHROMATOGRAPHY
seconde 2010 / 13: extraction/séparation / IV-1 chromatographie

COMMENTS

PDF Experiment Paper Chromatographic Separation and Identification of Sugars
In the last experiment you have separated metal ions by paper chromatography. In this experiment you will again to use pa.per chromatography for the separation of sugars. The term sugar applies to mono-, and ply-saccharides, which are all soluble in water and thereby distinguishable from plysaccharides. Many natural sugars are sweet.
PDF Chromatographic Separation of sugars
Chromatography is a technique for separation of closely related groups of compounds. The separation is brought about by differential migration along a porous medium and the migration is caused by the flow of solvent. Within limits chromatography can be divided into two types : partition and adsorption chromatography .Paper chromatography is an ...
PDF Paper Chromatography
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PDF Experiment 4
The chromatography paper should be dried and attached to your report. Procedure 1. Obtain a rectangular piece of chromatography paper. Use gloves to handle it. If gloves are not available, touch it only at the edges. (Oils from your hands can affect the separation.) 2. Make sure that the chromatography paper will fit into the beaker you plan to use
PDF Experiment 11 Paper Chromatography
Make sure the paper does not touch the sides of the beaker. Allow the solvent front to migrate up to 1 cm below the edge of the paper (top) for at least 90 minutes. Afterwards, remove the paper from the cylinder, mark the edge of the wet part of the paper, and allow it to air dry on the lab bench top.
PDF 11 PAPER CHROMATOGRAPHY
Remove the paper cylinders when the solvent reaches 1/2 1 cm from the top of the chromatography paper. Mark the highest point the solvent reached with a pencil and allow the paper to dry upright on a paper towel. When the paper is dry, remove the staples.
PDF EXPERIMENT 1: Thin-Layer Chromatography and Column Chromatography
Prior to performing a bulk separation and isolation using column chromatography, analysis of the plant extract by thin-layer chromatography (TLC) will be performed in order to find the best solvent system to be used in the large-scale separation. TLC with plant extracts produces several spots; easily identified by colour.
PDF PAPER CHROMATOGRAPHY
Obtain a strip of chromatography paper about 2.5 cm wide by 10 cm tall. Along one of the shorter sides, draw a horizontal line in pencil about 1.5 cm from the edge of the strip. This will be your "base line", the starting line where the samples will be spotted. Graphite will not be carried up the chromatography paper.
PDF Lab 2D: Separation of a Mixture by Paper Chromatography
a circular test in the lab. aphy piece of filter paper. Fold and cut to make. approx 20 cm by 12cm piece of unknowns of2. the paper3. spot the remaining 2 marks with your toothpick, by a 1mm spot so o. respective paper can ch.
PDF SEPARATION TECHNIQUES
1. Simple chromatography is carried out on paper. 2. A spot of the mixture is placed near the bottom of a piece of chromatography paper and the paper is then placed upright in a suitable solvent, eg water. 3. As the solvent soaks up the paper, it carries the mixtures with it. 4. Different components of the mixture will move at different rates. 5.
PDF Thin Layer Chromatography (TLC)
the paper and lead to erroneous results if it is touched on the "bottom". Lay the sheet of chromatography paper on a piece of notebook paper, and draw a line in pencil, not pen, 1.5 cm above the bottom. Make small marks along the line using the dimensions given in Figure 1 on the Figs. 1, Expt. 4 page (you may bring the figure pages to lab ...
PDF Separation and Analysis of Some Sugars by Using Thin Layer Chromatography
Leave to dry in. a1r for about 30 minutes. 6. Place the plate back in the same developing solvent and let the solvent move in the same. direc~ion. to the same d1stance of 12.5 cm. This usually takes 45 minutes. The ,plates should then be dried in air for apnroximately 30 m1nutes.
PDF Experiment 17: Chromatography
Chromatography is defined to be a chemical method of component separation where two. issolve. nd divide parts of aliquid. In its oldest and first uses, chromatography was used in the. widely-used until years laterwhen scientists discovered the method while. parating parts of.
Paper chromatography
Paper Chromatography Principle. The principle involved can be partition chromatography or adsorption chromatography. Partition chromatography because the substances are partitioned or distributed between liquid phases. The two phases are water held in pores of the filter paper and the other phase is a mobile phase which passes through the paper.
eGyanKosh: Experiment-5 Paper Chromatographic Seperation and
DSpace JSPUI eGyanKosh preserves and enables easy and open access to all types of digital content including text, images, moving images, mpegs and data sets
Separation of Sugars Using Paper Chromatography
4. Separation of Sugars Using Paper Chromatography..Docx - Free download as PDF File (.pdf), Text File (.txt) or read online for free. The document describes the procedure for separating sugars using paper chromatography, including the requirements, solvents, spray reagents, steps of the procedure, and results including calculating Rf values to identify unknown sugars in a mixture based on a ...
PDF EXPERIMENT 12 EXPERIMENTS BASED ON CHROMATOGRAPHY
check the purity of a compound using paper and thin layer chromatography. I 12.2 SEPARATION OF CONSTITUENTS OF A MIXTURE OF RED AND BLUE INKS BY PAPER CHROMATOGRAPHY This experiment is based upon paper chromatography. Paper chromatography is a very useful technique for separating mixtures of metal ions, anions, amino I acids, sugars, dyes ...
PDF Experiment 6
look like after development with solvent. (3) Use your draw. ng to show how the Rf value is determined.Thin-layer chromatography (TLC) is an extremely valuab. analytical technique in the organic lab. It provides a rapid separation of compounds, and thereby gives an indication of the number.
PDF Chromatography
With the help of a pencil, mark a line at a distance of 3 cm from one of the ends of this paper. (ii) Put a spot of the mixture on the marked line with the help of a fine capillary. (iii) Hang the filter paper in a jar containing a mixture of ethanol, (iv) 6.0 M HNO and distilled water, in the ratio 8:1:1. 3.
Separation and Identification of Sugars using Paper Chromatography
Abstract. CHROMATOGRAPHY on filter paper has provided an excellent and a much-needed method for the investigation of the structure of polysaccharides 1-5 and proteins 6-10. While butanol ...
Sustainability
A simple two-stage extraction and recovery method for macromolecules from microalgae biomass, termed CASS (concentrating the microalgae solution, acid pretreatment, high-shear-assisted lipid extraction, and separation), was developed. This method effectively processed the wet biomass of Chlorella sp. ABC-001 at a moderately low biomass concentration (50 g/L). The optimal conditions were acid ...
PDF Experiment Paper Chromatographic Separation and Identification of Metal
Next experiment concerned with PC separation of the sugars. 43.1 Principle In paper chromatography of cations, the principles of partition, adsorption and ion exchange may be exploited, out of these the most important is partition the involves the distribution of a solute between a mobile liquid phase and a gel (a kind of water

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