This document provides an overview of chromatographic techniques. It begins with definitions and a brief history, then covers principles, applications, classification, specific techniques (e.g. gas chromatography, liquid chromatography), terms, and properties of stationary phases. The document presents chromatographic methods and their use in separating mixtures like drugs, proteins, and other compounds. It concludes that supercritical fluid chromatography falls between HPLC and GC in performance for applications in pharmaceutical and bioanalytical analysis.
This document provides information about gas chromatography. It discusses the key components of a gas chromatography system including the mobile phase, stationary phase, columns, temperature control, sample injection systems, and various detectors. The mobile phase is usually an inert gas like helium, hydrogen, or nitrogen. Stationary phases can be solid adsorbents or liquid coatings. Columns include packed columns and capillary columns. Temperature control programs are used to separate compounds of varying boiling points. Common detectors mentioned are the thermal conductivity detector, flame ionization detector, and electron capture detector.
X-Ray Crystallography is a technique used to determine the atomic and molecular structure of a crystal, in which the crystalline atoms cause a beam of incident X-rays to diffract into many specific directions.
This form of chromatography is based on a thin film formed on the surface of a solid support by a liquid stationary phase. Solute equilibrates between the mobile phase and the stationary liquid. Components within a mixture are separated in a column based on each component's affinity for the mobile phase. If the components are of different polarities and a mobile phase of a distinct polarity is passed through the column, one component will migrate through the column faster than the other.
Affinity chromatography is a method used to purify biomolecules like proteins and nucleic acids based on specific interactions between the biomolecule and a ligand immobilized on a solid support. When a mixture is passed through the column, the target biomolecule will bind to the ligand while other molecules pass through. The bound biomolecule can then be separated by changing conditions like pH or introducing a competing molecule to displace it. Affinity chromatography offers highly specific purification of target molecules in a single step.
chromatography, principle, adsorbent of TLC, mobile phase of TLC, techniques in TLC, preparation of TLC plate, standards for TLC, advantages, disadvantages of TLC, Application of TLC.
This document provides an overview of gas chromatography. It begins with an introduction to chromatography and lists some common chromatographic techniques. It then describes the basic components and working of gas chromatography, including the carrier gas, columns, temperature control, detectors, and how the chromatographic process separates components based on partitioning between a mobile and stationary phase. The principle of gas chromatography is described as partition, with examples of different types of columns and factors that influence chromatographic separation. The key components of a gas chromatography system and their functions are also summarized.
The document discusses various chromatography techniques including thin layer chromatography (TLC), paper chromatography, and column chromatography. It provides details on the principles, instrumentation, advantages and applications of each technique. TLC involves spotting a sample on a plate coated with silica, developing it in a solvent, and visualizing results. Paper chromatography uses filter paper as the stationary phase. Column chromatography uses a glass column packed with an adsorbent like silica as the stationary phase.
This document provides information about gas chromatography. It discusses the key components of a gas chromatography system including the mobile phase, stationary phase, columns, temperature control, sample injection systems, and various detectors. The mobile phase is usually an inert gas like helium, hydrogen, or nitrogen. Stationary phases can be solid adsorbents or liquid coatings. Columns include packed columns and capillary columns. Temperature control programs are used to separate compounds of varying boiling points. Common detectors mentioned are the thermal conductivity detector, flame ionization detector, and electron capture detector.
X-Ray Crystallography is a technique used to determine the atomic and molecular structure of a crystal, in which the crystalline atoms cause a beam of incident X-rays to diffract into many specific directions.
This form of chromatography is based on a thin film formed on the surface of a solid support by a liquid stationary phase. Solute equilibrates between the mobile phase and the stationary liquid. Components within a mixture are separated in a column based on each component's affinity for the mobile phase. If the components are of different polarities and a mobile phase of a distinct polarity is passed through the column, one component will migrate through the column faster than the other.
Affinity chromatography is a method used to purify biomolecules like proteins and nucleic acids based on specific interactions between the biomolecule and a ligand immobilized on a solid support. When a mixture is passed through the column, the target biomolecule will bind to the ligand while other molecules pass through. The bound biomolecule can then be separated by changing conditions like pH or introducing a competing molecule to displace it. Affinity chromatography offers highly specific purification of target molecules in a single step.
chromatography, principle, adsorbent of TLC, mobile phase of TLC, techniques in TLC, preparation of TLC plate, standards for TLC, advantages, disadvantages of TLC, Application of TLC.
This document provides an overview of gas chromatography. It begins with an introduction to chromatography and lists some common chromatographic techniques. It then describes the basic components and working of gas chromatography, including the carrier gas, columns, temperature control, detectors, and how the chromatographic process separates components based on partitioning between a mobile and stationary phase. The principle of gas chromatography is described as partition, with examples of different types of columns and factors that influence chromatographic separation. The key components of a gas chromatography system and their functions are also summarized.
The document discusses various chromatography techniques including thin layer chromatography (TLC), paper chromatography, and column chromatography. It provides details on the principles, instrumentation, advantages and applications of each technique. TLC involves spotting a sample on a plate coated with silica, developing it in a solvent, and visualizing results. Paper chromatography uses filter paper as the stationary phase. Column chromatography uses a glass column packed with an adsorbent like silica as the stationary phase.
UPLC provides faster, more sensitive chromatographic separations compared to HPLC. It works by using smaller particle sizes (<2.5um) in the column packing which allows for higher pressure and flow rates based on the van Deemter equation. This provides benefits like reduced run times, decreased sample volume needs, and improved resolution. However, it also requires more robust instrumentation to handle the increased pressures and columns have reduced lifespan. UPLC has applications in fields like pharmaceutical analysis, metabolomics, and impurity profiling due to its enhanced resolution and sensitivity capabilities.
Paper chromatography is a technique used to separate mixtures based on differences in how components partition between a stationary phase (filter paper) and mobile phase (solvent). It involves applying a sample to a strip of paper and developing it by allowing the solvent to travel up the paper. This causes the different components to separate into bands at different distances based on how strongly they are retained by the paper. Paper chromatography is widely used for qualitative analysis and identification of organic and inorganic compounds in mixtures.
Paper chromatography is a method to separate components of a mixture using a stationary and mobile phase. It was introduced in 1865 and involves using specialized paper as the stationary phase and a solvent as the mobile phase. There are two main types - paper partition chromatography where the paper acts as the adsorbent, and paper adsorption chromatography where moisture in the paper acts as the stationary phase. Separation is mainly based on partition between the mobile and stationary phases. Different modes include ascending, descending, radial, and two-dimensional development depending on the solvent flow direction. The Rf value indicates the distance traveled by each component relative to the solvent front.
Chromatography is a technique used to separate and identify the components of a mixture. It works by allowing the molecules present in the mixture to distribute themselves between a stationary and a mobile medium. Molecules that spend most of their time in the mobile phase are carried along faster. There are different types of chromatography classified according to the mobile phase used and the packing of the stationary phase. Chromatography techniques include thin layer chromatography, paper chromatography, column chromatography, gas chromatography and liquid chromatography. These techniques find application in various fields to analyze sample mixtures.
G. Sateesh Chandra presented a seminar on gel chromatography under the guidance of Dr. P. Ramalingam. Gel chromatography, also known as size exclusion chromatography, separates dissolved molecules based on their size by pumping them through columns containing a microporous gel packing material. It is used to isolate and analyze biomacromolecules. The seminar covered the principles, instrumentation involving stationary phase, mobile phase, columns, pumps and detectors, methodology including sample preparation and elution, and applications such as group separation, molecular weight determination, and protein analysis.
This document provides an overview of chromatography techniques. It defines chromatography as a physical separation method that distributes components between two phases, one stationary and one mobile. It then classifies chromatography based on the stationary and mobile phases used as well as the instruments involved. Several chromatography techniques are described in detail, including thin layer chromatography. The principles, components, preparation, and considerations for thin layer chromatography are explained.
Adsorption chromatography is a technique for separating components in a mixture based on differential adsorption of the components onto a stationary solid phase. It works by passing a mobile liquid or gas phase over an adsorbent stationary phase in a column, which causes components to separate as they are differentially retained on the surface of the adsorbent. Common types include thin layer chromatography, paper chromatography, and column chromatography. Adsorption chromatography has various applications such as separating amino acids, isolating antibiotics, and identifying carbohydrates.
Introduction to gas Chromatography
,Principle of gas chromatography
Instrumentation of gas Chromatography
Type of detectors of gas chromatography
Advantages of gas chromatography
Disadvantages of gas chromatography
Applications of gas chromatography
High performance liquid chromatography is a powerful tool in analysis, it yields high performance and high speed compared to traditional columns chromatography because of the forcibly pumped mobile phase.
HPLC is a chromatographic technique that can separate a mixture of compounds.
The principle involved in HPLC can be either adsorption or partition.
This document provides an overview of partition chromatography. It defines partition chromatography as a method of separation where components in a mixture distribute between two immiscible liquid phases due to differences in their partition coefficients. The document discusses the history of chromatography and describes partition chromatography techniques like paper chromatography and gas-liquid chromatography. It explains the basic principles, procedures, factors affecting separation, and applications of partition chromatography.
Thin layer chromatography (TLC) is a technique used to separate mixtures of compounds and identify their components. It involves spotting a sample onto a thin layer of adsorbent material and using a mobile phase solvent to migrate the components at different rates based on their interactions with the stationary and mobile phases. TLC is useful for identifying unknown compounds, analyzing purity, and separating mixtures. It has advantages over column chromatography like being faster, using less solvent, and allowing detection of both colored and non-colored compounds.
Thin-layer chromatography (TLC) is a chromatography technique used to separate non-volatile mixtures. Thin-layer chromatography is performed on a sheet of glass, plastic, or aluminium foil, which is coated with a thin layer of adsorbent material, usually silica gel, aluminium oxide (alumina), or cellulose.
Capillary electrophoresis is a technique that uses narrow bore capillaries to separate charged molecules via electrophoretic mobility. When a voltage is applied, molecules migrate through the capillary at different rates depending on their charge and size. This allows analytes like proteins, nucleic acids, and small molecules to be separated. Key advantages are high efficiency, short analysis times, and low sample volume requirements. Common modes include capillary zone electrophoresis, capillary gel electrophoresis, and micellar electrokinetic capillary chromatography. Applications include analysis in pharmaceuticals and detection of microbial contamination.
This document discusses high performance thin layer chromatography (HPTLC). It begins with an introduction and overview of HPTLC principles, instrumentation, differences from TLC, and application steps. The document then provides more details on the HPTLC instrumentation components, factors affecting separation, common stationary and mobile phases used, and application techniques. It concludes by discussing detection methods and some key applications of HPTLC in pharmaceuticals, food analysis, clinical studies, and forensics.
Gas chromatography is a technique used to separate and analyze mixtures that can be vaporized without decomposition. It works by partitioning components to be separated between a stationary phase and a mobile gas phase. The key components of a gas chromatography instrument are the carrier gas, injection port, column, temperature control system, and detector. Factors like temperature, flow rate, column length, and amount of sample injected can influence separation of the components. Gas chromatography has applications in qualitative and quantitative analysis and is used in quality control of pharmaceuticals.
This document provides an overview of gas chromatography. It discusses the basic principles and components of gas chromatography including the stationary and mobile phases, how samples are injected and separated in the column based on their partitioning properties. Key components like the carrier gas, temperature control, detectors, and columns are described. The document outlines some parameters used to evaluate chromatography performance and lists common applications of gas chromatography in fields like pharmaceutical analysis, food testing, and environmental analysis.
This document provides an overview of gas chromatography. It discusses the basic components and principles of GC, including the mobile and stationary phases, instrumentation, and applications. The key points are:
- GC separates components in a sample using an inert gas mobile phase and either a solid or liquid stationary phase in the column.
- Common instrumentation includes the carrier gas, flow regulators, sample injector, temperature-controlled column oven, detectors, and recorders.
- Separation is based on how strongly components partition between the mobile and stationary phases.
- GC has wide applications in fields like pharmaceutical analysis, environmental analysis, forensics, and industrial quality control.
- Advantages are strong separation power,
Gel permeation chromatography is a size-exclusion chromatography technique that separates analytes based on their hydrodynamic size. Molecules are separated by passing through a column containing porous gel or polymer beads. Small molecules are able to enter the pores and thus remain inside the beads longer, while larger molecules pass through more quickly. Detectors such as refractive index and UV-Vis are used to analyze the separated analyte bands exiting the column. GPC can determine molecular weight distribution and is used to analyze polymers, proteins, and other large biomolecules.
The document discusses different types of chromatography. It begins with an introduction to chromatography, including its history and principles. It then describes various classifications of chromatography based on mechanism and phases. Specific techniques are defined, including adsorption chromatography, partition chromatography, gas-liquid chromatography, solid-liquid chromatography, and liquid-liquid chromatography. Key terms are explained. Applications and steps of chromatographic separation are outlined. Important properties of liquid stationary phases are also summarized.
UPLC provides faster, more sensitive chromatographic separations compared to HPLC. It works by using smaller particle sizes (<2.5um) in the column packing which allows for higher pressure and flow rates based on the van Deemter equation. This provides benefits like reduced run times, decreased sample volume needs, and improved resolution. However, it also requires more robust instrumentation to handle the increased pressures and columns have reduced lifespan. UPLC has applications in fields like pharmaceutical analysis, metabolomics, and impurity profiling due to its enhanced resolution and sensitivity capabilities.
Paper chromatography is a technique used to separate mixtures based on differences in how components partition between a stationary phase (filter paper) and mobile phase (solvent). It involves applying a sample to a strip of paper and developing it by allowing the solvent to travel up the paper. This causes the different components to separate into bands at different distances based on how strongly they are retained by the paper. Paper chromatography is widely used for qualitative analysis and identification of organic and inorganic compounds in mixtures.
Paper chromatography is a method to separate components of a mixture using a stationary and mobile phase. It was introduced in 1865 and involves using specialized paper as the stationary phase and a solvent as the mobile phase. There are two main types - paper partition chromatography where the paper acts as the adsorbent, and paper adsorption chromatography where moisture in the paper acts as the stationary phase. Separation is mainly based on partition between the mobile and stationary phases. Different modes include ascending, descending, radial, and two-dimensional development depending on the solvent flow direction. The Rf value indicates the distance traveled by each component relative to the solvent front.
Chromatography is a technique used to separate and identify the components of a mixture. It works by allowing the molecules present in the mixture to distribute themselves between a stationary and a mobile medium. Molecules that spend most of their time in the mobile phase are carried along faster. There are different types of chromatography classified according to the mobile phase used and the packing of the stationary phase. Chromatography techniques include thin layer chromatography, paper chromatography, column chromatography, gas chromatography and liquid chromatography. These techniques find application in various fields to analyze sample mixtures.
G. Sateesh Chandra presented a seminar on gel chromatography under the guidance of Dr. P. Ramalingam. Gel chromatography, also known as size exclusion chromatography, separates dissolved molecules based on their size by pumping them through columns containing a microporous gel packing material. It is used to isolate and analyze biomacromolecules. The seminar covered the principles, instrumentation involving stationary phase, mobile phase, columns, pumps and detectors, methodology including sample preparation and elution, and applications such as group separation, molecular weight determination, and protein analysis.
This document provides an overview of chromatography techniques. It defines chromatography as a physical separation method that distributes components between two phases, one stationary and one mobile. It then classifies chromatography based on the stationary and mobile phases used as well as the instruments involved. Several chromatography techniques are described in detail, including thin layer chromatography. The principles, components, preparation, and considerations for thin layer chromatography are explained.
Adsorption chromatography is a technique for separating components in a mixture based on differential adsorption of the components onto a stationary solid phase. It works by passing a mobile liquid or gas phase over an adsorbent stationary phase in a column, which causes components to separate as they are differentially retained on the surface of the adsorbent. Common types include thin layer chromatography, paper chromatography, and column chromatography. Adsorption chromatography has various applications such as separating amino acids, isolating antibiotics, and identifying carbohydrates.
Introduction to gas Chromatography
,Principle of gas chromatography
Instrumentation of gas Chromatography
Type of detectors of gas chromatography
Advantages of gas chromatography
Disadvantages of gas chromatography
Applications of gas chromatography
High performance liquid chromatography is a powerful tool in analysis, it yields high performance and high speed compared to traditional columns chromatography because of the forcibly pumped mobile phase.
HPLC is a chromatographic technique that can separate a mixture of compounds.
The principle involved in HPLC can be either adsorption or partition.
This document provides an overview of partition chromatography. It defines partition chromatography as a method of separation where components in a mixture distribute between two immiscible liquid phases due to differences in their partition coefficients. The document discusses the history of chromatography and describes partition chromatography techniques like paper chromatography and gas-liquid chromatography. It explains the basic principles, procedures, factors affecting separation, and applications of partition chromatography.
Thin layer chromatography (TLC) is a technique used to separate mixtures of compounds and identify their components. It involves spotting a sample onto a thin layer of adsorbent material and using a mobile phase solvent to migrate the components at different rates based on their interactions with the stationary and mobile phases. TLC is useful for identifying unknown compounds, analyzing purity, and separating mixtures. It has advantages over column chromatography like being faster, using less solvent, and allowing detection of both colored and non-colored compounds.
Thin-layer chromatography (TLC) is a chromatography technique used to separate non-volatile mixtures. Thin-layer chromatography is performed on a sheet of glass, plastic, or aluminium foil, which is coated with a thin layer of adsorbent material, usually silica gel, aluminium oxide (alumina), or cellulose.
Capillary electrophoresis is a technique that uses narrow bore capillaries to separate charged molecules via electrophoretic mobility. When a voltage is applied, molecules migrate through the capillary at different rates depending on their charge and size. This allows analytes like proteins, nucleic acids, and small molecules to be separated. Key advantages are high efficiency, short analysis times, and low sample volume requirements. Common modes include capillary zone electrophoresis, capillary gel electrophoresis, and micellar electrokinetic capillary chromatography. Applications include analysis in pharmaceuticals and detection of microbial contamination.
This document discusses high performance thin layer chromatography (HPTLC). It begins with an introduction and overview of HPTLC principles, instrumentation, differences from TLC, and application steps. The document then provides more details on the HPTLC instrumentation components, factors affecting separation, common stationary and mobile phases used, and application techniques. It concludes by discussing detection methods and some key applications of HPTLC in pharmaceuticals, food analysis, clinical studies, and forensics.
Gas chromatography is a technique used to separate and analyze mixtures that can be vaporized without decomposition. It works by partitioning components to be separated between a stationary phase and a mobile gas phase. The key components of a gas chromatography instrument are the carrier gas, injection port, column, temperature control system, and detector. Factors like temperature, flow rate, column length, and amount of sample injected can influence separation of the components. Gas chromatography has applications in qualitative and quantitative analysis and is used in quality control of pharmaceuticals.
This document provides an overview of gas chromatography. It discusses the basic principles and components of gas chromatography including the stationary and mobile phases, how samples are injected and separated in the column based on their partitioning properties. Key components like the carrier gas, temperature control, detectors, and columns are described. The document outlines some parameters used to evaluate chromatography performance and lists common applications of gas chromatography in fields like pharmaceutical analysis, food testing, and environmental analysis.
This document provides an overview of gas chromatography. It discusses the basic components and principles of GC, including the mobile and stationary phases, instrumentation, and applications. The key points are:
- GC separates components in a sample using an inert gas mobile phase and either a solid or liquid stationary phase in the column.
- Common instrumentation includes the carrier gas, flow regulators, sample injector, temperature-controlled column oven, detectors, and recorders.
- Separation is based on how strongly components partition between the mobile and stationary phases.
- GC has wide applications in fields like pharmaceutical analysis, environmental analysis, forensics, and industrial quality control.
- Advantages are strong separation power,
Gel permeation chromatography is a size-exclusion chromatography technique that separates analytes based on their hydrodynamic size. Molecules are separated by passing through a column containing porous gel or polymer beads. Small molecules are able to enter the pores and thus remain inside the beads longer, while larger molecules pass through more quickly. Detectors such as refractive index and UV-Vis are used to analyze the separated analyte bands exiting the column. GPC can determine molecular weight distribution and is used to analyze polymers, proteins, and other large biomolecules.
The document discusses different types of chromatography. It begins with an introduction to chromatography, including its history and principles. It then describes various classifications of chromatography based on mechanism and phases. Specific techniques are defined, including adsorption chromatography, partition chromatography, gas-liquid chromatography, solid-liquid chromatography, and liquid-liquid chromatography. Key terms are explained. Applications and steps of chromatographic separation are outlined. Important properties of liquid stationary phases are also summarized.
This document provides an overview of different chromatographic methods. It discusses the basic principles of chromatography and defines key terms. It then classifies chromatography based on mechanism of separation (adsorption vs partition) and phases (solid, liquid, gas). Several specific chromatographic techniques are described in more detail, including gas-liquid chromatography, solid-liquid chromatography, and liquid-liquid chromatography. The document also discusses planar chromatography techniques like paper chromatography and thin layer chromatography as well as column chromatography. Important properties of liquid stationary phases are also outlined.
This document provides an overview of chromatography. It defines chromatography as a set of laboratory techniques used to separate mixtures based on how components partition between a mobile and stationary phase. The document then classifies chromatography based on mechanism, phases used, and shape of the chromatographic bed. It proceeds to describe various chromatography techniques in more detail, including adsorption chromatography, partition chromatography, gas-liquid chromatography, solid-liquid chromatography, and column chromatography.
Chromatography is a method of separating mixtures into individual components using a stationary and mobile phase. There are several types depending on the physical state of the phases and interaction between the phases and components. Liquid chromatography uses a liquid mobile phase passing through a solid or liquid stationary phase to separate components. Gas chromatography uses a gas mobile phase to separate volatile components. Size exclusion and ion exchange chromatography separate based on molecular size or charge.
Chromatography is a technique used to separate mixtures by distributing components between two phases, stationary and mobile. The mixture is dissolved in a mobile phase that carries it through a column containing a stationary phase. Components travel at different rates based on how they partition between the phases, allowing separation. Common chromatography methods include gas chromatography, liquid chromatography, and thin layer chromatography. Chromatography has applications in identifying unknown substances like drugs, proteins, and plant pigments. It was first developed in 1903 and continues to be an important analytical technique.
Chromatography is a technique used to separate mixtures based on how their components interact with both a mobile and stationary phase. It was first developed in 1900 by Russian scientist Mikhail Tsvet to separate plant pigments. There are several types of chromatography that differ based on the phases used, including paper chromatography, thin layer chromatography, gas chromatography, ion exchange chromatography, gel filtration chromatography, and affinity chromatography. High performance liquid chromatography is a modern technique that uses small particle sizes and high pressure to improve separation efficiency.
This document provides information about chromatography. It defines chromatography as a method of separation where components are distributed between a stationary and mobile phase. The stationary phase can be solid or liquid, and the mobile phase can be liquid, gas, or supercritical fluid. Various types of chromatography are described based on the interaction between components and phases, including thin layer chromatography, column chromatography, gas chromatography, and liquid chromatography. Key applications and principles of different chromatographic techniques are also summarized.
Chromatography is a separation technique that was discovered in 1906 by Russian scientist Mikhail Tswett. It involves distributing components of a mixture between a stationary and mobile phase. There are different types of chromatography based on the phases used, such as gas chromatography which uses a gas mobile phase, and liquid chromatography which uses a liquid mobile phase. Chromatography can also be classified based on the mechanism of separation, such as adsorption chromatography where components are absorbed onto a solid stationary phase.
Chromatography is a technique used to separate mixtures. It works by dissolving the mixture in a mobile phase that carries it through a structure containing a stationary phase. The different constituents of the mixture travel at different speeds depending on how they interact with the mobile and stationary phases, resulting in separation. There are various types of chromatography classified based on the physical state of the mobile and stationary phases, including adsorption chromatography where separation occurs via differences in adsorption to the stationary phase, partition chromatography based on differences in solubility between phases, and thin layer chromatography using a thin stationary phase coated sheet.
Chromatography is a laboratory technique used to separate components of a mixture based on how they interact with mobile and stationary phases. It was first developed in 1901 by Russian botanist Mikhail Tswett to separate plant pigments. The components move through the stationary phase at different rates, allowing separation. Chromatography has important analytical and preparative uses and involves terms like chromatograph, eluent, eluate, stationary phase, and mobile phase.
Introduction to chromatography and its applications 2Kalsoom Mohammed
Chromatography is a technique used to separate mixtures based on differences in how components interact with stationary and mobile phases. The document defines chromatography and describes its history, principles, commonly used terms, types including adsorption (gas chromatography, thin layer chromatography, column chromatography, ion exchange chromatography, HPLC) and partition (paper chromatography, gas chromatography), working, detectors, visualization, applications and references. Chromatography is widely used in fields like pharmaceuticals, food, forensics and more to analyze and purify chemical mixtures.
Chromatography is a technique used to separate mixtures based on how their components interact with a mobile and stationary phase. The mixture dissolves in a mobile phase that carries it through a structure containing a stationary phase. Components travel at different speeds depending on how they partition between the mobile and stationary phases, resulting in separation. Chromatography has various applications like analyzing drugs, proteins, and complex mixtures. Common types include paper, thin layer, gas-liquid, and high performance liquid chromatography.
This document provides an introduction to chromatography, including its history and essential features. It discusses the basic components and process of chromatography, including the stationary and mobile phases. It also describes different types of chromatography techniques based on the stationary phase, such as partition chromatography, adsorption chromatography, ion exchange chromatography, molecular exclusion chromatography, and affinity chromatography. Finally, it discusses applications of chromatography in qualitative analysis, quantitative analysis, and preparative purposes.
Chromatography is a technique used to separate mixtures by distributing components between two phases - a stationary phase and a mobile phase. The document discusses the history, principles, types (including adsorption, partition, thin layer, gas, and high performance liquid), applications, and terminology of chromatography. Key types are paper chromatography, gas chromatography, and HPLC. Chromatography is widely used in industries like pharmaceuticals and food to analyze compounds.
• Chromatography is a method of separation in which the components to be separated are distributed between two phases, one of these is called a stationary phase and the other is a mobile phase which moves on stationary phase in a definite direction
The document discusses different types of chromatography techniques used to separate mixtures. It describes chromatography as a collective term for laboratory methods that separate mixtures by dissolving them in a mobile phase that carries the mixture through a stationary phase. The key types discussed are paper chromatography, thin layer chromatography, gas chromatography, column chromatography, and high performance liquid chromatography. HPLC uses very small column packing and high pressure to improve separation speed over other chromatography methods.
Chromatography is a method used to separate mixtures by distributing components between a stationary and mobile phase. There are several types including thin layer chromatography (TLC), which separates compounds on plates coated with adsorbents, and column chromatography, where the stationary phase is packed in a tube. High performance liquid chromatography (HPLC) uses pumps to force liquid mobile phases through columns at high pressures for improved separation.
Chromatography is a method used to separate mixtures by distributing components between a stationary and mobile phase. There are several types including thin layer chromatography (TLC), which separates compounds on plates coated with adsorbent. Column chromatography packs an absorbent in a column and elutes components with a solvent. High performance liquid chromatography (HPLC) uses high pressure to force components through a column at high speed. Chromatography techniques are used in forensic analysis and research to identify substances.
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2. CONTENTS
Introduction to chromatography
History
Principles
Importance
Chromatographic terms
Classification of chromatography
Adsorption chromatography
Partition chromatography
Gas-liquid phase chromatrography
Solid-liquid phase chromatrography
Liquid-gas phase chromatrography
Liquid-liquid phase chromatrography
Important properties of liquid phase
Conclusion
2
3. Chromatography
Chromatography (from Greek chroma "color
and graphein "to write") is the collective term for
a set of laboratory techniques for the separation
of mixtures.
The mixture is dissolved in a fluid called the
mobile phase, which carries it through a
structure holding another material called the
stationary phase.
The various constituents of the mixture travel at
different speeds, causing them to separate. The
separation is based on differential partitioning
between the mobile and stationary phases.
3
4. History
Chromatography, literally "color writing",
was first employed by Russian scientist
Mikhail Tsvet in 1900.
He continued to work with chromatography
in the first decade of the 20th century,
primarily for the separation of plant
pigments such as chlorophyll, carotenes,
and xanthophylls.
Since these components have different
colors (green, orange, and
yellow,respectively) they gave the
technique its name.
4
5. Principles
Chromatography usually consists of
mobile phase and stationary phase. The
mobile phase refers to the mixture of
substances to be separated dissolved in
a liquid or a gas.
The stationary phase is a porous solid
matrix through which the sample
contained in the mobile phase
percolates.
The interaction between the mobile
phase and the stationary phase results in
the separation of the compound from the
5
6. Applications of
chromatography
The chromatographic technique is used
for the separation of amino
acids,proteins & carbohydrates.
It is also used for the analysis of
drugs,hormones,vitamins.
Helpful for the qualitative & quantitative
analysis of complex mixtures.
The technique is also useful for the
determination of molecular weight of
proteins.
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7. The chromatographic method
of separation, in general,
involves following steps
Adsorption or retention of substances on the
stationary phase
Separation of the adsorption of substances by
the mobile phase
Recovery of the separated substances by a
continuous flow of the mobile phase; the
method being called elution
Qualitative and Qantitative analysis of the
eluted substances
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8. Chromatographic terms
The analyte is the substance to be separated during
chromatography.
A chromatogram is the visual output of the
chromatograph.
The eluate is the mobile phase leaving the column.
The eluent is the solvent that carries the analyte
The detector refers to the instrument used for
qualitative and quantitative detection of analytes after
separation.
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9. Classification of chromatography
1. Based on mechanism of separation
I. adsorption chromatography
II. Partition chromatography
2. Based on phases
I. Solid phase chromatography
i. Solid-liquid chromatography
ii. Solid-gas chromatography
II. Liquid phase chromatography
i. Liquid-liquid chromatography
ii. Liquid –gas chromatography
3. Based on shape of chromatographic bed
I. Planner chromatography
i. Paper chromatography
ii. Thin layer chromatography
II. Column chromatography
i. Packed column chromatography
ii. Open tubular column chromatography
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10. Flow chart diagram of chromatography
chromatography
adsorption
Competition between
Solid and
Gas
(G.S.C.)
Liquid
Column
chromatography
Thin layer
chromatography
partition
Competition between
Liquid and
Gas
G.L.C.
Liquid
H.P.L.C.
Column
chromatography
Paper
chromatography
TLC
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11. Adsorption chromatograohy
Adsorption chromatography is one of the oldest types of
chromatography.
It utilizes a mobile liquid or
gaseous phase that is
adsorbed onto the surface of
a stationary solid phase
The equilibriation between the mobile and stationary
phase accounts for the separation of different solutes.
Adsorption chromatography is process of separation of
components in a mixture introduced into chromatography
system based on the relative difference in adsorption of
components to stationary phase present in chromatography
column
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sumit prajapati
12. Partition chromatography
This form of chromatography is based on a thin film formed
on the surface of a solid support by a liquid stationary phase
Solute equilibrates
between the mobile phase
and the stationary liquid.
Chromatography in which separation is based mainly on
differences between the solubility of the sample components in
the stationary phase or on differences between the solubility of
the components in the mobile and stationary phases
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sumit prajapati
13. Gas-Solid chromatography(G.S.C.)
Gas chromatography employs an inert gas as the mobile
phase
Separation depends on the relative partial pressures of
the sample components above the stationary phase.
Gas-solid chromatography is relatively rare, but it is used
to separate atmospheric gases
Common solids are charcoal, a synthetic zeolite called
"molecular sieve", or a combination of the two.
The mobile phase is a gas, often nitrogen, but
sometimes helium, hydrogen or occasionally another gas.
It is called the "carrier gas".
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14. Solid-Liquid chromatography
Liquid chromatography (LC) is a separation technique
in which the mobile phase is a liquid.
Liquid chromatography can be carried out either in a
column or a plane
In liquid-solid chromatography the porous adsorbent
is polar and separation is based on the properties of
classes of compounds—e.g., amines (alkaline) from
alcohols (neutral) and esters (neutral) from acids
The preferred mobile phase is a nonpolar or slightly
polar...
Popular adsorbents are Silica and Alumina.
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15. Liquid-Gas Chromatography
Dimethyl Polysiloxane (350oC)
Hydrocarbons, Polynuclear aromatics
Poly(phenyl methyl) siloxane (250oC)
Steroids, Pesticides, Glycols
Stationary phase used in (LGC)
The mobile phase is an unreactive gas, such as nitrogen
(the carrier gas)
The stationary phase comprises of a small amount of
liquid held on a finely-divided inert solid support.
Gas-liquid chromatography is very sensitive and can be
used to detect small quantities of substances
it is often used in forensic tests
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16. Liquid-Liquid Chromatography
Liquid-liquid chromatography is a chromatography
separation technique in which the mobile phase is a liquid
(usually a solvent or a simple binary solvent mixture) and
the stationary phase is also a liquid (which must be
immiscible and insoluble in the liquid mobile phase).
The first liquid-liquid system was reported by A. J. P.
Martin who used water supported on silica gel as the
stationary phase and n-heptane as the mobile phase
The system is inherently unstable, as the stationary
phase will always have some solubility in mobile phase
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17. Planner chromatography
Planar chromatography is a separation technique in
which the stationary phase is present on a plane.
The plane can be a paper, serving as such or
impregnated by a substance as the stationary bed (paper
chromatography) or a layer of solid particles spread on a
support such as a glass plate (Thin layer
chromatography).
Different compounds in the sample mixture travel
different distances according to how strongly they interact
with the stationary phase as compared to the mobile
phase.
The specific Retention factor (Rf) of each chemical can
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18. Column Chromatography
Column chromatography is a separation technique in
which the stationary bed is within a tube.
The particles of the solid stationary phase or the
support coated with a liquid stationary phase may fill the
whole inside volume of the tube (packed column) or be
concentrated on or along the inside tube wall leaving an
open, unrestricted path for the mobile phase in the middle
part of the tube (open tubular column).
Differences in rates of movement through the medium
are calculated to different retention times of the sample
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19. Important properties of liquid
stationary phase
Liquid phase should have low volatility and high stability
at elevated temperatures
Liquid phase should not permeate too deeply into the
fine pores of the support structure as slow diffusion in and
out of pores affects column efficiency
Small particles of support give higher efficiency as
HETP is proportional to particle diameter but particle size
reduction increases back pressure
Support should be deactivated before use as
undesirable surface impurities can cause decomposition of
the sample or stationary liquid
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20. Conclusion
In overall ranking Chromatography
techniques , it can be judge SFC falls
somewhere between HPLC or GC.
In field of pharmaceutical chemistry and
bioanalytical application gained its
applications
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