The document discusses different types of chromatography techniques. It explains that ion exchange chromatography separates ions based on their electrostatic attraction to charged groups on a resin stationary phase. Size exclusion chromatography separates molecules based on their size, with larger molecules passing through bead pores faster. Affinity chromatography uses a ligand immobilized on a stationary phase to separate molecules based on specific binding interactions. The document also classifies chromatography based on mobile phase, stationary phase polarity, and the shape of the chromatographic bed. It describes uses of chromatography for qualitative analysis to identify components and quantitative analysis to determine concentrations.
• 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
This document discusses the eight main types of chromatography: paper chromatography, thin layer chromatography, gel chromatography, column chromatography, ion exchange chromatography, gel filtration chromatography, gas liquid chromatography, and affinity chromatography. Each type is briefly described in one or two sentences explaining the basic components and separation process used.
Chromatography and its classifications along with principles and applications...Sukanta Debnath
The document provides an overview of various chromatography techniques including thin layer chromatography (TLC), gas chromatography (GC), high performance thin layer chromatography (HPTLC), column chromatography, ion exchange chromatography, gel filtration chromatography, high performance liquid chromatography (HPLC), and affinity chromatography. It discusses the basic principles, applications, and key steps for each technique. The document is intended as a presentation on modern pharmaceutical analytical chromatography techniques.
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with a stationary and mobile phase. It was developed in 1906 and is widely used in science and industry. The main types are paper, thin layer, gas, and liquid chromatography. Chromatography works by differential partitioning or adsorption of components between the mobile and stationary phases as they move through a column or plate. This allows separation based on properties like size, charge, or binding affinity. It has many applications like analyzing pharmaceuticals, foods, forensics samples, and more.
This document provides information on chromatography techniques. It discusses the basic components and processes involved in chromatography. The key techniques described are column chromatography, planar chromatography, gas chromatography, liquid chromatography, and affinity chromatography. It also summarizes different modes of chromatography such as normal phase chromatography, reverse phase chromatography, ion exchange chromatography, size exclusion chromatography, and affinity chromatography.
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.
The document discusses various analytical chromatography techniques. It describes chromatography as separating components through distribution between two immiscible phases, with one stationary and one mobile. The document outlines different types of chromatography including column chromatography, thin layer chromatography, gas chromatography, and ion exchange chromatography. It discusses the principles, techniques, and efficiency of these analytical methods.
This document discusses various downstream processing techniques used to purify proteins produced by biopharmaceutical manufacturing. It describes key steps like removal of insolubles, product isolation, purification, and polishing. Specific techniques covered include chromatography methods like ion exchange, size exclusion, hydrophobic interaction, and affinity chromatography. It also provides examples of using hydrophobic interaction chromatography and affinity chromatography to purify green fluorescent protein.
• 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
This document discusses the eight main types of chromatography: paper chromatography, thin layer chromatography, gel chromatography, column chromatography, ion exchange chromatography, gel filtration chromatography, gas liquid chromatography, and affinity chromatography. Each type is briefly described in one or two sentences explaining the basic components and separation process used.
Chromatography and its classifications along with principles and applications...Sukanta Debnath
The document provides an overview of various chromatography techniques including thin layer chromatography (TLC), gas chromatography (GC), high performance thin layer chromatography (HPTLC), column chromatography, ion exchange chromatography, gel filtration chromatography, high performance liquid chromatography (HPLC), and affinity chromatography. It discusses the basic principles, applications, and key steps for each technique. The document is intended as a presentation on modern pharmaceutical analytical chromatography techniques.
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with a stationary and mobile phase. It was developed in 1906 and is widely used in science and industry. The main types are paper, thin layer, gas, and liquid chromatography. Chromatography works by differential partitioning or adsorption of components between the mobile and stationary phases as they move through a column or plate. This allows separation based on properties like size, charge, or binding affinity. It has many applications like analyzing pharmaceuticals, foods, forensics samples, and more.
This document provides information on chromatography techniques. It discusses the basic components and processes involved in chromatography. The key techniques described are column chromatography, planar chromatography, gas chromatography, liquid chromatography, and affinity chromatography. It also summarizes different modes of chromatography such as normal phase chromatography, reverse phase chromatography, ion exchange chromatography, size exclusion chromatography, and affinity chromatography.
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.
The document discusses various analytical chromatography techniques. It describes chromatography as separating components through distribution between two immiscible phases, with one stationary and one mobile. The document outlines different types of chromatography including column chromatography, thin layer chromatography, gas chromatography, and ion exchange chromatography. It discusses the principles, techniques, and efficiency of these analytical methods.
This document discusses various downstream processing techniques used to purify proteins produced by biopharmaceutical manufacturing. It describes key steps like removal of insolubles, product isolation, purification, and polishing. Specific techniques covered include chromatography methods like ion exchange, size exclusion, hydrophobic interaction, and affinity chromatography. It also provides examples of using hydrophobic interaction chromatography and affinity chromatography to purify green fluorescent protein.
Chromatography is a technique used to separate mixtures into individual components. It works by distributing components between a stationary and mobile phase based on differences in how strongly each component interacts with the phases. The main types are adsorption, partition, ion exchange, size exclusion, and affinity chromatography. Chromatography is widely used in industries like pharmaceuticals, food, chemicals, and molecular biology to analyze, identify, purify, and quantify mixtures and components.
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.
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 the components of a mixture. There are several types of chromatography based on the mechanism of separation, including ion-exchange, affinity, size-exclusion, adsorption, and partition chromatography. The document provides definitions and principles of each type as well as examples of their applications in separating biochemical compounds like proteins, peptides, amino acids, and alkaloids. Chromatography is widely used in fields like biochemistry, forensic science, and environmental analysis.
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 laboratory technique used to separate mixtures into individual components. It works by distributing the components between two phases, usually a stationary phase and a mobile phase. There are several types of chromatography defined by the stationary and mobile phases used, including gas chromatography which uses an inert gas as the mobile phase, and high performance liquid chromatography which uses high pressure to force a liquid mobile phase through a column. Chromatography has many applications in fields like chemistry and biochemistry for analyzing and purifying compounds.
Advanced techniques and laborotory equipments for biologistsNawfal Aldujaily
Gas chromatography (GC) separates volatile compounds using an inert gas as the mobile phase. The sample is injected into a heated port to volatilize it. The gas mobile phase carries the volatilized sample through a heated column coated with a stationary phase that interacts with analytes. Components are separated based on differences in volatility and affinity for the stationary phase, then detected and recorded. GC is useful for separating volatile, non-polar compounds.
Chromatography is based on the principle where molecules in mixture applied onto the surface or into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase.
The factors effective on this separation process include molecular characteristics related to adsorption (liquid-solid), partition (liquid-solid), and affinity or differences among their molecular weights
Because of these differences, some components of the mixture stay longer in the stationary phase, and they move slowly in the chromatography system, while others pass rapidly into mobile phase, and leave the system faster.
Chromatography is a physical separation method that separates components of a mixture based on how they interact with different phases. There are various terms used in chromatography including the stationary phase, mobile phase, analyte, and retention time. Chromatography techniques can be classified based on their mechanism of separation, including ion-exchange chromatography, affinity chromatography, size-exclusion chromatography, adsorption chromatography, and partition chromatography. Each technique utilizes different interactions like charge, binding affinity, size, or partitioning to separate substances. Chromatography has many applications like protein purification, water analysis, and quality control.
Chromatography is a physical separation method that separates components of a mixture based on how they interact with different phases. There are various terms used in chromatography including the stationary phase, mobile phase, analyte, and retention time. Chromatography techniques can be classified based on their mechanism of separation, including ion-exchange chromatography, affinity chromatography, size-exclusion chromatography, adsorption chromatography, and partition chromatography. Each technique utilizes different interactions like charge, binding affinity, size, or partitioning to separate substances. Chromatography has many applications like protein purification, water analysis, and quality control.
Chromatography is a physical separation method that separates components of a mixture based on how they interact with different phases. There are various terms used in chromatography including the stationary phase, mobile phase, analyte, and retention time. Chromatography techniques can be classified based on their mechanism of separation, including ion-exchange chromatography, affinity chromatography, size-exclusion chromatography, adsorption chromatography, and partition chromatography. Each technique utilizes different interactions like charge, binding affinity, size, or partitioning to separate substances. Chromatography has many applications like protein purification, water analysis, and quality control.
This document provides an overview of high-performance liquid chromatography (HPLC). It begins by defining HPLC and explaining that it uses small particle columns and high pressure to achieve faster separations compared to traditional liquid chromatography. The document then discusses the basic components and principles of HPLC, including the stationary and mobile phases, various modes of separation, and common instrumentation such as pumps, injectors, columns, and detectors. It provides details on the configuration and function of each component.
HPLC - High Performance Liquid ChromatographyDivya Basuti
The document discusses High Performance Liquid Chromatography (HPLC). It explains that HPLC is a type of liquid chromatography that uses pumps to force the mobile phase through a column packed with porous particles or beads under high pressure. This allows for effective separation of mixtures as the components elute from the column at different rates depending on their interactions with the stationary phase. The document provides details on the typical components of an HPLC system including the solvent delivery system, pumps, injector, columns, detectors, and data processing unit.
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, paper chromatography, gas chromatography, and liquid chromatography. The separation is based on differences in how components partition between the phases and interact with the stationary phase material. Chromatography is widely used to identify unknown substances and test purity.
Chromatography is a technique used to separate mixtures by distributing components between two phases - a stationary phase and a mobile phase. Mikhail Tswett discovered chromatography in 1906 when separating plant pigments. There are various types of chromatography classified by mobile phase (liquid, gas), stationary phase material (thin layer, paper, column), or separation mechanism (adsorption, partition, ion exchange, size exclusion, affinity). Chromatography has many applications in science and industry, including purification of antibiotics, vaccines, enzymes, and other biomolecules.
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.
This document provides an overview of chromatography. It discusses the history and discovery of chromatography by Tswett in 1906. It then defines chromatography and describes the basic components of a chromatogram. The document classifies chromatography by mobile and stationary phase as well as by separation mechanism. It discusses various chromatography techniques including thin layer chromatography, column chromatography, gas chromatography, and high performance liquid chromatography. It also covers separation factors such as solute retention, capacity factor, and efficiency.
1. Asifa Bibi presented on various chromatography techniques. She discussed paper chromatography, column chromatography, gel filtration chromatography, ion exchange chromatography, and affinity chromatography.
2. The key principles of chromatography are the separation of mixtures based on differences in how components partition between a stationary and mobile phase.
3. Chromatography techniques have many applications in fields like biotechnology, pharmaceuticals, and forensic analysis.
Chromatography is a technique used to separate mixtures by differential distribution between a stationary and mobile phase. It has various applications in science and medicine. The document discusses the history of chromatography, types including thin layer chromatography, gas chromatography, high performance liquid chromatography, and modes such as adsorption, partition, ion exchange, and size exclusion chromatography. Chromatography is commonly used for analytical and preparative purposes in various fields.
Chromatography is a technique used to separate mixtures into individual components. It works by distributing components between a stationary and mobile phase based on differences in how strongly each component interacts with the phases. The main types are adsorption, partition, ion exchange, size exclusion, and affinity chromatography. Chromatography is widely used in industries like pharmaceuticals, food, chemicals, and molecular biology to analyze, identify, purify, and quantify mixtures and components.
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.
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 the components of a mixture. There are several types of chromatography based on the mechanism of separation, including ion-exchange, affinity, size-exclusion, adsorption, and partition chromatography. The document provides definitions and principles of each type as well as examples of their applications in separating biochemical compounds like proteins, peptides, amino acids, and alkaloids. Chromatography is widely used in fields like biochemistry, forensic science, and environmental analysis.
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 laboratory technique used to separate mixtures into individual components. It works by distributing the components between two phases, usually a stationary phase and a mobile phase. There are several types of chromatography defined by the stationary and mobile phases used, including gas chromatography which uses an inert gas as the mobile phase, and high performance liquid chromatography which uses high pressure to force a liquid mobile phase through a column. Chromatography has many applications in fields like chemistry and biochemistry for analyzing and purifying compounds.
Advanced techniques and laborotory equipments for biologistsNawfal Aldujaily
Gas chromatography (GC) separates volatile compounds using an inert gas as the mobile phase. The sample is injected into a heated port to volatilize it. The gas mobile phase carries the volatilized sample through a heated column coated with a stationary phase that interacts with analytes. Components are separated based on differences in volatility and affinity for the stationary phase, then detected and recorded. GC is useful for separating volatile, non-polar compounds.
Chromatography is based on the principle where molecules in mixture applied onto the surface or into the solid, and fluid stationary phase (stable phase) is separating from each other while moving with the aid of a mobile phase.
The factors effective on this separation process include molecular characteristics related to adsorption (liquid-solid), partition (liquid-solid), and affinity or differences among their molecular weights
Because of these differences, some components of the mixture stay longer in the stationary phase, and they move slowly in the chromatography system, while others pass rapidly into mobile phase, and leave the system faster.
Chromatography is a physical separation method that separates components of a mixture based on how they interact with different phases. There are various terms used in chromatography including the stationary phase, mobile phase, analyte, and retention time. Chromatography techniques can be classified based on their mechanism of separation, including ion-exchange chromatography, affinity chromatography, size-exclusion chromatography, adsorption chromatography, and partition chromatography. Each technique utilizes different interactions like charge, binding affinity, size, or partitioning to separate substances. Chromatography has many applications like protein purification, water analysis, and quality control.
Chromatography is a physical separation method that separates components of a mixture based on how they interact with different phases. There are various terms used in chromatography including the stationary phase, mobile phase, analyte, and retention time. Chromatography techniques can be classified based on their mechanism of separation, including ion-exchange chromatography, affinity chromatography, size-exclusion chromatography, adsorption chromatography, and partition chromatography. Each technique utilizes different interactions like charge, binding affinity, size, or partitioning to separate substances. Chromatography has many applications like protein purification, water analysis, and quality control.
Chromatography is a physical separation method that separates components of a mixture based on how they interact with different phases. There are various terms used in chromatography including the stationary phase, mobile phase, analyte, and retention time. Chromatography techniques can be classified based on their mechanism of separation, including ion-exchange chromatography, affinity chromatography, size-exclusion chromatography, adsorption chromatography, and partition chromatography. Each technique utilizes different interactions like charge, binding affinity, size, or partitioning to separate substances. Chromatography has many applications like protein purification, water analysis, and quality control.
This document provides an overview of high-performance liquid chromatography (HPLC). It begins by defining HPLC and explaining that it uses small particle columns and high pressure to achieve faster separations compared to traditional liquid chromatography. The document then discusses the basic components and principles of HPLC, including the stationary and mobile phases, various modes of separation, and common instrumentation such as pumps, injectors, columns, and detectors. It provides details on the configuration and function of each component.
HPLC - High Performance Liquid ChromatographyDivya Basuti
The document discusses High Performance Liquid Chromatography (HPLC). It explains that HPLC is a type of liquid chromatography that uses pumps to force the mobile phase through a column packed with porous particles or beads under high pressure. This allows for effective separation of mixtures as the components elute from the column at different rates depending on their interactions with the stationary phase. The document provides details on the typical components of an HPLC system including the solvent delivery system, pumps, injector, columns, detectors, and data processing unit.
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, paper chromatography, gas chromatography, and liquid chromatography. The separation is based on differences in how components partition between the phases and interact with the stationary phase material. Chromatography is widely used to identify unknown substances and test purity.
Chromatography is a technique used to separate mixtures by distributing components between two phases - a stationary phase and a mobile phase. Mikhail Tswett discovered chromatography in 1906 when separating plant pigments. There are various types of chromatography classified by mobile phase (liquid, gas), stationary phase material (thin layer, paper, column), or separation mechanism (adsorption, partition, ion exchange, size exclusion, affinity). Chromatography has many applications in science and industry, including purification of antibiotics, vaccines, enzymes, and other biomolecules.
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.
This document provides an overview of chromatography. It discusses the history and discovery of chromatography by Tswett in 1906. It then defines chromatography and describes the basic components of a chromatogram. The document classifies chromatography by mobile and stationary phase as well as by separation mechanism. It discusses various chromatography techniques including thin layer chromatography, column chromatography, gas chromatography, and high performance liquid chromatography. It also covers separation factors such as solute retention, capacity factor, and efficiency.
1. Asifa Bibi presented on various chromatography techniques. She discussed paper chromatography, column chromatography, gel filtration chromatography, ion exchange chromatography, and affinity chromatography.
2. The key principles of chromatography are the separation of mixtures based on differences in how components partition between a stationary and mobile phase.
3. Chromatography techniques have many applications in fields like biotechnology, pharmaceuticals, and forensic analysis.
Chromatography is a technique used to separate mixtures by differential distribution between a stationary and mobile phase. It has various applications in science and medicine. The document discusses the history of chromatography, types including thin layer chromatography, gas chromatography, high performance liquid chromatography, and modes such as adsorption, partition, ion exchange, and size exclusion chromatography. Chromatography is commonly used for analytical and preparative purposes in various fields.
8 Surprising Reasons To Meditate 40 Minutes A Day That Can Change Your Life.pptxHolistified Wellness
We’re talking about Vedic Meditation, a form of meditation that has been around for at least 5,000 years. Back then, the people who lived in the Indus Valley, now known as India and Pakistan, practised meditation as a fundamental part of daily life. This knowledge that has given us yoga and Ayurveda, was known as Veda, hence the name Vedic. And though there are some written records, the practice has been passed down verbally from generation to generation.
Integrating Ayurveda into Parkinson’s Management: A Holistic ApproachAyurveda ForAll
Explore the benefits of combining Ayurveda with conventional Parkinson's treatments. Learn how a holistic approach can manage symptoms, enhance well-being, and balance body energies. Discover the steps to safely integrate Ayurvedic practices into your Parkinson’s care plan, including expert guidance on diet, herbal remedies, and lifestyle modifications.
TEST BANK For An Introduction to Brain and Behavior, 7th Edition by Bryan Kol...rightmanforbloodline
TEST BANK For An Introduction to Brain and Behavior, 7th Edition by Bryan Kolb, Ian Q. Whishaw, Verified Chapters 1 - 16, Complete Newest Versio
TEST BANK For An Introduction to Brain and Behavior, 7th Edition by Bryan Kolb, Ian Q. Whishaw, Verified Chapters 1 - 16, Complete Newest Version
TEST BANK For An Introduction to Brain and Behavior, 7th Edition by Bryan Kolb, Ian Q. Whishaw, Verified Chapters 1 - 16, Complete Newest Version
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TEST BANK For Basic and Clinical Pharmacology, 14th Edition by Bertram G. Katzung, Verified Chapters 1 - 66, Complete Newest Version.
TEST BANK For Basic and Clinical Pharmacology, 14th Edition by Bertram G. Katzung, Verified Chapters 1 - 66, Complete Newest Version.
TEST BANK For Basic and Clinical Pharmacology, 14th Edition by Bertram G. Katzung, Verified Chapters 1 - 66, Complete Newest Version.
TEST BANK For Basic and Clinical Pharmacology, 14th Edition by Bertram G. Katzung, Verified Chapters 1 - 66, Complete Newest Version.
Basavarajeeyam is a Sreshta Sangraha grantha (Compiled book ), written by Neelkanta kotturu Basavaraja Virachita. It contains 25 Prakaranas, First 24 Chapters related to Rogas& 25th to Rasadravyas.
Muktapishti is a traditional Ayurvedic preparation made from Shoditha Mukta (Purified Pearl), is believed to help regulate thyroid function and reduce symptoms of hyperthyroidism due to its cooling and balancing properties. Clinical evidence on its efficacy remains limited, necessitating further research to validate its therapeutic benefits.
REGULATION FOR COMBINATION PRODUCTS AND MEDICAL DEVICES.pptx
chromatography 2 (2).pptx
1. PHARMACOGNSY-IIA (Advance) [Theory]
Course No. 513-T(Morn)
Course Incharge: Ms. Mehwish Khan
Chapter: Separation and isolation of plant
constituents
Topic: Separation mechanism in different chromatography
2. Previous lecture
Following points have been discussed in the previous lecture,
Chromatography is a separation technique in which a sample is
equilibrated between two immiscible phase (mobile and stationary
phase).
Separation mechanism in different types of chromatography depends
mainly on the nature of the stationary phase
Adsorption chromatography, mixture of gas or liquid gets separated
when it passes over the adsorbent bed that adsorbs different
compounds at different rates
Partition chromatography is process of separation whereby the
components of the mixture get distributed into two liquid phases due
to differences in partition coefficients
3. OBJECTIVE
• In this lecture, we will study about
classification of chromatography i.e. ion
exchange, size exclusion and affinity
chromatography and also classification
according to mobile phase, polarity & shape
of chromatographic bed.
4. “Ion exchange chromatography may be defined
as the reversible exchange of ions in the solution with
ions electrostatically bound to stationary phase.”
The stationary phase is an ion exchange resin to which a cationic or anionic
groups are covalently bonded.
Ions of opposite charges (counter ions) in the mobile phase will be attracted
to the resin and compete with the components of the mixture for the
charged group on the resin.
Consider column having E - Y+ cation
exchanger in which E - is negative
charged exchanger and Y+ is the mobile
counter ion. X+ be the cation in the
sample having charge greater than Y+.
The X+ ion can exchange sites with the
counter ion Y+
5. Ion – Exchange chromatography
Bounded interest of ion (X+ ) can now
be eluted by either of the two ways;
1. By adding a component M+ having
magnitude of charge more than
that of X+ so that M+ will replace
X+ and X+ will be eluting out.
2. By changing pH of the solvent
(mobile phase) to neutralize the
component X+ have no charge and is
then unbounded from the matrix and
can be eluted out.
6. Molecular Exclusion or Gel filtration chromatography
• Size-exculsion chromatography (SEC), also called gel filtration or gel-
permeation chromatography (GPC), uses porous particles to separate
molecules of different sizes.
Stationary phase used for gel flitration / size exclusion chromatography
macromolecular complexes include dextran (Sephadex), polyacrylamide
and dextranpolyacrylamide (Sephacryl).
• Each is available with a variety of different ranges of pore size in the
beads, permitting separation of macromolecules of different size
• A mixture of molecules dissolved in liquid (the mobile phase) is applied to
a chromatography column which contains a solid support in the form of
microscopic spheres, or “beads” (the stationary phase).
• Size of pores in beads determines the exclusion limit (what goes through
the beads and what goes around the beads)
7. Working of gel-permeation chromatography Larger molecules pass around
or are “excluded” from the beads .
• Large sample molecules cannot or can only partially penetrate the pores,
whereas smaller molecules can access most or all pores.
• Thus, large molecules elute first, smaller molecules elute later, while
molecules that can access all the pores elute last from the column.
• Particles of different sizes will elute (filter) through a stationary phase at
different rates.
8. Affinity Chromatography
• Affinity chromatography is a method of separating biochemical
mixtures based on a highly specific interaction such as that between
antigen and antibody, enzyme and substrate, or receptor and ligand
• It is a method of separating a mixture of proteins or nucleic acids
(molecules) by specific interactions of those molecules with a
component known as a ligand, which is immobilized on a support.
• If a solution or mixture of proteins is passed over (through) the
column, one of the proteins binds to the ligand on the basis of
specificity and high affinity (they fit together like a lock and key).
• The other proteins in the solution wash through the column because
they were not able to bind to the ligand.
• The ligand/molecule complex dissociates by changing the pH.
9. Affinity Chromatography
APPLICATIONS
• Production of Vaccines -
antibody purification from
blood serum
• Used in Genetic Engineering -
nucleic acid purification
• Basic Metabolic Research -
protein or enzyme purification
from cell free extracts
10. ACCORDING TO MOBILE PHASE AND MECHANISM
Liquid Chromatography (LC)
• The mobile phase is liquid. In case of separation by adsorption the stationary
phase is solid so it is called: Liquid-Solid Chromatography (LSC).
• If separation occurs through partition, the stationary phase is liquid so it is
called: Liquid –Liquid Chromatography (LLC).
Gas Chromatography (GC)
• Where the mobile phase is inert gas nitrogen or helium. Again if in case of
separation by adsorption, the stationary phase is solid it is called: Gas–Solid
Chromatography (GSC).
• If separation occurs through partition, the stationary phase is liquid it is
called: Gas-Liquid Chromatography (GLC).
ACCORDING TO POLARITY OF PHASES
Liquid Chromatography (LC)
• The stationary phase is more polar and the mobile phase is less polar so it is called normal
phase chromatography
• The stationary phase is less polar and the mobile phase is more polar so it is called
reverese phase chromatography
11. According to the technique
(methods of holding the stationary phase)
Planar or Plane Chromatography:
• In this type of chromatography the stationary phase is used in the
form of layer. Plane chromatography is further classified into:
a- Thin Layer Chromatography (TLC):
• A technique used to separate non-volatile mixtures. It is performed
on a sheet of glass, plastic, or aluminum foil, which is coated with a
thin layer of adsorbent material, usually silica gel, aluminium oxide
(alumina), or cellulose.
b- Paper Chromatography (PC):
• A specific type of papers is used as stationary phase in the form of
sheets.
Columnar or Column Chromatography (CC):
• The stationary phase is held in to a tube made of glass or metal.
12. Analytical Chromatography
• Chromatography can be used to obtain pure materials from mixtures
anddetermine the existence and also the concentration of analyte(s)
materials in a sample.
Qualitative Chromatography
• Confirm the absence or presence of certain constituent in the sample
• Thin-layer chromatography (TLC) is a widely used method for qualitative
analysis to determine the number of components in a mixture, to
determine the identity of substances in sample
Quantitative Chromatography
• Quantitative chromatography is used to determine the concentration of
analytes in a sample.
• The components is identified by its retention time and concentration
calculated from intensity of detector signal.
• HPLC/ GC can be used for these applications.
13. Summary
Ion exchange chromatography may be defined as the reversible exchange
of ions in the solution with ions electrostatically bound to stationary
phase. Bounded ions of sample can be eluted according to their binding
strength to resin by two ways.
• Size-exculsion chromatography (SEC), also called gel filtration or gel-
permeation chromatography (GPC), uses porous particles to separate
molecules of different sizes
• Affinity chromatography is a method of separating biochemical mixtures
based on a highly specific interaction
• Chromatography cab be classified according to mobile phase and shape of
chromatographic bed on which separation of molecules occurred.
• Qualitative Chromatography Confirm the absence or presence of certain
constituent in the sample and Quantitative chromatography is used to
determine the concentration of analytes in a sample
14. Further reading and references
• Chromatographic Methods. A. Braithwaite and F.J. Smith.
Published by Kluwer Academic. Publisher
• The Essence of chromatography. Colin F. Poole. by Elsevier