2.3.2
Thin-Layer Chromatography
Thin-Layer Chromatography
Thin-Layer Chromatography
Thin-layer chromatography (TLC) is a type of chromatography used to identify and separate chemicals in a mixture.
 8.1.3.1 - Paper chromatography apparatus,h_400,q_80,w_640.jpg)
 8.1.3.1 - Paper chromatography apparatus,h_400,q_80,w_640.jpg)
1) Setup the apparatus
1) Setup the apparatus
- Use an aluminium TLC plate coated in silica or alumina.
- The substances to be separated or identified must be dissolved in organic solvent and dotted along the bottom of the TLC plate.
- The TLC plate is placed in a closed jar with a small amount of organic solvent inside.
 8.1.3.1 - Paper chromatography apparatus,h_400,q_80,w_640.jpg)
 8.1.3.1 - Paper chromatography apparatus,h_400,q_80,w_640.jpg)
2) Run the experiment
2) Run the experiment
- The jar is left and the organic solvent in the jar rises up the TLC plate and over the dots.
- The TLC plate is removed when the solvent front (top of the rising solvent on the plate) nears the top edge of the plate.
- We can now use this TLC plate to either identify or separate the components in the mixture.


3) Evaluate the results
3) Evaluate the results
- The mobile phase is the organic solvent (typically apolar).
- The stationary phase is the TLC plate (typically polar).
- The more polar substances will interact more with the stationary phase and so will be found lower down the plate.
- The less polar substances will interact more with the mobile phase and so will rise with the solvent more quickly and will be found higher up the plate.
Separation and Identification
Separation and Identification
TLC can be used to separate and identify substances in mixtures.
 _Diagram 14.8.3.3b - and subsequent identification of amino acids by chromatography. ,h_400,q_80,w_640.png)
 _Diagram 14.8.3.3b - and subsequent identification of amino acids by chromatography. ,h_400,q_80,w_640.png)
Separation
Separation
- When separation is required, substances from the mixture can be isolated by cutting up regions with just one spot from the TLC plate.
 8.1.3.2 - chromatogram,h_400,q_80,w_640.jpg)
 8.1.3.2 - chromatogram,h_400,q_80,w_640.jpg)
Rf values
Rf values
- We use Rf values to identify substances. These values are calculated as:
- Rf = distance travelled by substance รท distance travelled by solvent
- We can calculate the Rf value for reference substances and the tested substances.
 8.1.3.2 - chromatogram,h_400,q_80,w_640.jpg)
 8.1.3.2 - chromatogram,h_400,q_80,w_640.jpg)
Identification
Identification
- Rf values depend on the solvent that they are measured in.
- The identification process is comparing the Rf value of the reference and tested substance.


Calculation of Rf
Calculation of Rf
- We have a mixture and we suspect that it contains a particular component e.g. propanol.
- Since Rf values are relative to the conditions, we must only compare values measured under the same conditions.
- The best way to do this is to add a spot of the suspected component, e.g. propanol, to the same TLC plate as the mixture and run them together, then we can see if the Rf values align.
- If the Rf values do align, there's significant evidence that the suspected component is in the mixture.
Colourless Chemicals
Colourless Chemicals
Some chemicals used in TLC can't be identified because they aren't visible on the TLC plate. The following are methods for locating these colourless chemicals:


Fluorescent dyes
Fluorescent dyes
- Fluorescent dyes are visible when UV light is shone on them.
- This allows us to see where the spot of the colourless chemical is and so we can calculate its Rf value.


Iodine
Iodine
- Iodine turns the colourless spots purple.
- This is another method that allows us to see where the spot of the colourless chemical is so that we can calculate its Rf value.
1Principles of Science I
1.1Structure & Bonding
1.1.1Atomic Model
1.1.2Electron Shells, Sub-Shells & Orbitals
1.1.3Ionic Bonding
1.1.4Representing Ionic Bonds
1.1.5Covalent Bonding
1.1.6Representing Covalent Bonds
1.1.7Metallic Bonding
1.1.8Intermolecular Forces
1.1.9Intermolecular Forces 2
1.1.10End of Topic Test - Bonding
1.1.11Relative Masses
1.1.12The Mole
1.1.13Molar Calculations
1.1.14Molar Calculations 2
1.1.15Empirical & Molecular Formulae
1.1.16Balanced Equations
1.1.17Percentage Yield
1.1.18End of Topic Test - Amount of Substance
1.2Properties of Substances
1.2.1The Periodic Table
1.2.2Ionisation Energy
1.2.3Factors Affecting Ionisation Energies
1.2.4Trends of Ionisation
1.2.5Trends in the Periodic Table
1.2.6Polarity
1.2.7Metals & Non-Metals
1.2.8Alkali Metals
1.2.9Alkaline Earth Metals
1.2.10Reactivity of Alkaline Earth Metals
1.2.11Redox
1.2.12Transition Metals
1.2.13Redox Reactions of Transition Metals
1.3Cell Structure & Function
1.4Cell Specialisation
1.5Tissue Structure & Function
1.5.1Human Gas Exchange
1.5.2Blood Vessels
1.5.3Atherosclerosis
1.5.4Skeletal Muscle
1.5.5Slow & Fast Twitch Fibres
1.5.6Neurones
1.5.7Speed of Transmission
1.5.8Action Potentials
1.5.9End of Topic Test - Neurones & Action Potentials
1.5.10Synapses
1.5.11Types of Synapse
1.5.12Medical Application
1.5.13End of Topic Test - Synapses
1.5.14Chemical Brain Imbalances
1.5.15Effect of Drugs on the Brain
1.6Working with Waves
1.7Waves in Communication
2Practical Scientific Procedures and Techniques
3Science Investigation Skills
3.1Scientific Processes
3.2Data Handling & Analysis
3.3Enzymes in Action
3.4Diffusion
3.5Plants & Their Environment
3.6Energy Content in Fuels
4Principles of Science II
4.1Extracting Elements
4.2Relating Properties to use of Substances
4.3Organic Chemistry
4.4Energy Changes in Industry
4.5The Circulatory System
4.5.1The Circulatory System
4.5.2Blood Vessels
4.5.3Blood Transfusion & the ABO Rhesus System
4.5.4The Heart
4.5.5The Cardiac Cycle
4.5.6Cardiac Output
4.5.7Coordination of Heart Action
4.5.8Heart Dissection
4.5.9Controlling Heart Rate
4.5.10Electrocardiograms
4.5.11Cardiovascular Disease
4.5.12Investigating Heart Rates
4.6Ventilation & Gas Exchange
4.7Urinary System
4.9Thermal Physics
4.9.1Power & Efficiency
4.9.2Work & Energy
4.9.3Conservation of Energy
4.9.4Pressure
4.9.5First Law of Thermodynamics
4.9.6Second Law of Thermodynamics
4.9.7Heat Engines, Heat Pumps & Refrigerators
4.9.8Non-Flow Processes
4.9.9p-V Diagrams
4.9.10Ideal Gases
4.9.11Ideal Gases 2
4.9.12Thermal Energy Transfer
4.9.13Thermal Energy Transfer Experiments
4.10Materials
5Contemporary Issues in Science
5.1Contemporary Issues in Science
5.2Analysing Scientific Information
Jump to other topics
1Principles of Science I
1.1Structure & Bonding
1.1.1Atomic Model
1.1.2Electron Shells, Sub-Shells & Orbitals
1.1.3Ionic Bonding
1.1.4Representing Ionic Bonds
1.1.5Covalent Bonding
1.1.6Representing Covalent Bonds
1.1.7Metallic Bonding
1.1.8Intermolecular Forces
1.1.9Intermolecular Forces 2
1.1.10End of Topic Test - Bonding
1.1.11Relative Masses
1.1.12The Mole
1.1.13Molar Calculations
1.1.14Molar Calculations 2
1.1.15Empirical & Molecular Formulae
1.1.16Balanced Equations
1.1.17Percentage Yield
1.1.18End of Topic Test - Amount of Substance
1.2Properties of Substances
1.2.1The Periodic Table
1.2.2Ionisation Energy
1.2.3Factors Affecting Ionisation Energies
1.2.4Trends of Ionisation
1.2.5Trends in the Periodic Table
1.2.6Polarity
1.2.7Metals & Non-Metals
1.2.8Alkali Metals
1.2.9Alkaline Earth Metals
1.2.10Reactivity of Alkaline Earth Metals
1.2.11Redox
1.2.12Transition Metals
1.2.13Redox Reactions of Transition Metals
1.3Cell Structure & Function
1.4Cell Specialisation
1.5Tissue Structure & Function
1.5.1Human Gas Exchange
1.5.2Blood Vessels
1.5.3Atherosclerosis
1.5.4Skeletal Muscle
1.5.5Slow & Fast Twitch Fibres
1.5.6Neurones
1.5.7Speed of Transmission
1.5.8Action Potentials
1.5.9End of Topic Test - Neurones & Action Potentials
1.5.10Synapses
1.5.11Types of Synapse
1.5.12Medical Application
1.5.13End of Topic Test - Synapses
1.5.14Chemical Brain Imbalances
1.5.15Effect of Drugs on the Brain
1.6Working with Waves
1.7Waves in Communication
2Practical Scientific Procedures and Techniques
3Science Investigation Skills
3.1Scientific Processes
3.2Data Handling & Analysis
3.3Enzymes in Action
3.4Diffusion
3.5Plants & Their Environment
3.6Energy Content in Fuels
4Principles of Science II
4.1Extracting Elements
4.2Relating Properties to use of Substances
4.3Organic Chemistry
4.4Energy Changes in Industry
4.5The Circulatory System
4.5.1The Circulatory System
4.5.2Blood Vessels
4.5.3Blood Transfusion & the ABO Rhesus System
4.5.4The Heart
4.5.5The Cardiac Cycle
4.5.6Cardiac Output
4.5.7Coordination of Heart Action
4.5.8Heart Dissection
4.5.9Controlling Heart Rate
4.5.10Electrocardiograms
4.5.11Cardiovascular Disease
4.5.12Investigating Heart Rates
4.6Ventilation & Gas Exchange
4.7Urinary System
4.9Thermal Physics
4.9.1Power & Efficiency
4.9.2Work & Energy
4.9.3Conservation of Energy
4.9.4Pressure
4.9.5First Law of Thermodynamics
4.9.6Second Law of Thermodynamics
4.9.7Heat Engines, Heat Pumps & Refrigerators
4.9.8Non-Flow Processes
4.9.9p-V Diagrams
4.9.10Ideal Gases
4.9.11Ideal Gases 2
4.9.12Thermal Energy Transfer
4.9.13Thermal Energy Transfer Experiments
4.10Materials
5Contemporary Issues in Science
5.1Contemporary Issues in Science
5.2Analysing Scientific Information
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