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Thin Layer Chromatography Investigations Leighton Dann and Paul Beaumont Science & Plants for Schools www.saps.org.uk
TLC One of a number of types of chromatography: gas/liquid affinity paper
TLC Separation of complex mixtures between two phases: -a stationary phase -and a mobile phase
TLC Phases the stationary phase is a thin layer of absorbent particles, in this case silica gel the mobile phase is the solvent
Fisher Scientific UK. Tel: 01509 555500 Cat No: TLC-410-610X
Thin Layer Chromatography Separation of: photosynthetic pigments in leaves anthocyanins in leaves, flowers, fruit and vegetables
Method Place ca. 3 cm 2 leaf in mortar [If tissue is coarse, silver sand may be added] Grind until all tissue broken up Add ca. 750 l propanone Pour off supernatant into a container and seal
Method Supernatant and residue from grinding leaf in propanone
Method Mark (with a pencil) a plate 1.5 cm up from the base on the edge of both sides Use a ruler across these marks to mark two spots 2 cm apart in the middle of the plate
Method Use Pasteur pipette to place small drops of supernatant Dry off before adding another lot of drops Continue until there is a concentrated line of colour
Method Repeat process until a second line is produced (or one line across the whole plate)
Method Plate into a 600 cm 3 beaker, replace foil Remove plate when solvent nearly at top Immediately mark solvent front
Solvent system Cyclohexane : propanone : petroleum ether (low boiling point) 5 : 3 : 2
PigmentColourR F value caroteneyellow-orange0.91 pheophytin agrey0.75 pheophytin blight grey0.63-0.75 chlorophyll ablue green0.63 chlorophyll bgreen0.58 xanthophyllsyellow0.53 xanthophyllsyellow0.47 xanthophyllsyellow0.32
S EPARATION BY CHROMATOGRAPHY Describe chromatography as an analytical technique that separates components in a mixture. State that the mobile phase may.
Describe the processes involved in the extraction and separation of photosynthetic pigments from a leaf extraction grind leaves in fine sand - to rupture.
Paper Chromatography Chromatography is a method of separating and identifying various components in a mixture, which are present in small trace quantities.
Experiment.13 Amino acid analysis by adsorption thin layer chromatography (adsorption TLC)
PAG6 TLC. Need to book Repro – Example results or Page 19 of delivery guide – Example method – FLIPPED learning sheet for start of nucleotides Applied.
Dr. Samah Kotb Nasr Eldeen. Amino acids: are the building blocks of peptides and proteins. They possess two functional groups, the carboxylic acid.
Alexandria University Faculty Of Science Practical Training Course In Applications Of Plant Physiology Prepared By Dr. Mohamed Mohamed Ibrahim Lecturer.
Cellular Biochemistry and Metabolism (CLS 333 ) Dr. Samah Kotb Nasr Eldeen Identification of free amino acids by Thin layer chromotography (TLC) using.
Chromatography Separating plant pigments. Cellulose Paper (stationary phase)
What is Chromatography? Chromatography is a technique for separating mixtures into their components in order to analyze, identify, purify, and/or quantify.
Today: Exp.6: Column Chromatography: The Separation of Ferrocene and Acetyl Ferrocene Midterm next Friday in your lab! starting at the beginning of your.
Thin-Layer Chromatography (TLC) Uses: To separate the components of a mixture To determine the purity of a compound To see if two compounds are identical.
1 Pigments in plants Separation of chloroplast pigments by paper chromatography Refer to the Practical Manual Unit 5: Cell Biology Practical 16.
Amino acids: are the building blocks of peptides and proteins. They possess two functional groups—the carboxylic acid group gives the acidic character,
Chemistry Project: Acids and alkalis Other details Experiment Results Web Site ‘s version The aim of the project: To know more the details of alkalis and.
By Susan McCullough With Thanks to Lori Olson at SRI International.
CHROMATOGRAPHY Lab # 5. introduction Chromatography is a laboratory technique for the separation of mixtures. It involves passing a mixture dissolved.
Experiment 15 A: Isolation of Pure Aspirin From Aspirin Tablets B: TLC of a Dye Mixture.
Chromatography. Chromatography is the collective term for a set of laboratory techniques used to separate mixtures. It involves passing a mixture dissolved.
Amanda Matsumura Nathan Marcy Alisa Craig Tim Waldrip Dr. Ellen Turner, Instructor.
PAPER CHROMATOGRAPHY AND THIN LAYER CHROMATOGRAPHY.
Pigment Chromatography. Plant leaves contain different color pigments that give the leaf color. Plant pigments come in many different colors but we are.
Thin layer chromatography (TLC) is an important technique for identification and separation of mixtures of organic compounds. It is useful in: Identification.
High Sucrose Soybean Thin-Layer Chromatography (TLC)
Paper Chromatography of a Spinach Leaf Lab *record all of the info underined and in red in your notebook Background Information: Paper chromatography.
0 Chromatography is a method of physically separating mixtures of gases, liquids, or dissolved substances. Chromatography can be used to identify drugs,
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T.A Nouf Alshareef KAU-Faculty of Science- Biochemistry department Analytical biochemistry lab (Bioc 343) 2012
Using chromatography to identify amino acids Key words: SOLVENT – a liquid in which chemicals dissolve to make a solution (e.g. water, ethanol) SOLUTE.
CHM 101/102 Laboratory Manual Chromatography I: Paper General Chemistry 101/102 Laboratory Manual University of North Carolina at Wilmington.
My Experiment Color Chromatography Presented By Kelsey Leigh Campbell Date: 3 October 2012.
Paper Chromatography Identifying the components of a mixture.
Chromatography and Instrumentation. Invented by a Russian Botanist Mikhail Tswett in 1903 He used chromatography to separate the colour pigments in plants.
T HIN - LAYER CHROMATOGRAPHY State that the mobile phase in TLC is a liquid and that the stationary phase is a solid on a solid support. State that the.
Extracting Chlorophyll Using paper chromatography The chemical formula for water = H2OH2O Hydrogen = H H2OH2O H 2 = 2 atoms of Hydrogen Oxygen = O.
Absorption Spectrum for Plant Pigments For this experiment you will be using a spectrophotometer to determine the combined absorption spectrum for the.
Chromatography is a technique for separating mixtures into their components in order to analyze, identify, purify, and/or quantify the mixture or components.
Chromo: color Graph: to write In the mid 1900s used to separate and analyze naturally occurring pigments like those in leaves. Chromatography year7hawkesdale.wikispaces.com.
1/17/ Thin-Layer Chromatography References: Pavia:Experiment 5A; pp. 36 – 39 Technique 2.0; pp. 801 – 812 Note:The above reference material is not.
Photosynthesis ‘Life is a tiny electric current kept moving by the sun’ Svent Gorki.
Analysing Substances LO: Describe how you would detect artificial food colourings (D) Explain how colourings can be identified using chromatography. (C)
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Chromatography Chapter Dr Gihan Gawish. 1. Paper Chromatography Dr Gihan Gawish Paper chromatography is a technique that involves placing a small.
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