Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with stationary and mobile phases. There are several types of chromatography that differ based on the phases used, including paper, column, thin layer, gas, high performance liquid, and affinity chromatography. Chromatography has many applications in fields like forensics, environmental testing, and drug analysis.
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.
Chromatography is a laboratory technique used to separate mixtures by distributing components to separate between two phases, one stationary and one mobile. It works based on how substances partition between the phases and move through the system at different rates. Common techniques include column chromatography, thin layer chromatography, gas chromatography, and high performance liquid chromatography. Chromatography is used in analytical chemistry to identify unknown substances and quantify components in a mixture.
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.
Gas liquid chromatography (GLC) is a technique where gaseous samples are separated into components through partition between a gaseous mobile phase (such as helium or nitrogen) and a liquid stationary phase held in a column. The mobile phase transports sample vapors through the column without chemical interaction. Components are separated due to differences in how they partition between the mobile and stationary phases. A detector then generates a signal proportional to solute concentration to produce a chromatogram. Factors like particle size, carrier gas flow rate, column properties, and temperature affect the separation. GLC allows both qualitative and quantitative analysis of many organic compounds.
Chromatography is a technique used to separate and identify the components of a mixture. It works by allowing molecules to distribute themselves between a stationary and mobile phase, so that molecules that interact more with the mobile phase move faster. Chromatographic techniques can be classified based on the interaction with the stationary phase or physical state of the mobile phase. Key techniques include adsorption, partition, ion exchange, exclusion, gas, liquid, and thin layer chromatography. Proper sample preparation and development conditions are important for achieving optimal separation and resolution of components in the mixture.
Chromatography is a scientific technique used to separate mixtures based on how compounds interact with two phases - a stationary phase and a mobile phase. There are several types of chromatography including thin layer chromatography, gas chromatography, high performance liquid chromatography, electrophoresis, and paper chromatography. Each type uses different stationary and mobile phases and has various applications such as determining compound compositions, analyzing organic reactions, and identifying unknown substances.
The document discusses the process and history of paper chromatography. It begins with an introduction to chromatography and its use in separating mixtures. It then covers the history from early experiments in the 1860s to modern developments. The main types and techniques of paper chromatography are explained, including the use of a stationary phase, mobile phase, and capillary action to separate components by affinity and travel distance. Key steps like sample application and developing the paper strip are outlined.
The document describes planar chromatography techniques, specifically thin layer chromatography (TLC). It explains that TLC separates mixtures by using a thin stationary phase like silica gel coated on a plate and a mobile phase liquid solvent. The steps of TLC are described as sample application, development where separation occurs, visualization under UV light, and interpretation by calculating Rf values. Applications for separating lipids, carbohydrates and other compounds are outlined.
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.
Chromatography is a laboratory technique used to separate mixtures by distributing components to separate between two phases, one stationary and one mobile. It works based on how substances partition between the phases and move through the system at different rates. Common techniques include column chromatography, thin layer chromatography, gas chromatography, and high performance liquid chromatography. Chromatography is used in analytical chemistry to identify unknown substances and quantify components in a mixture.
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.
Gas liquid chromatography (GLC) is a technique where gaseous samples are separated into components through partition between a gaseous mobile phase (such as helium or nitrogen) and a liquid stationary phase held in a column. The mobile phase transports sample vapors through the column without chemical interaction. Components are separated due to differences in how they partition between the mobile and stationary phases. A detector then generates a signal proportional to solute concentration to produce a chromatogram. Factors like particle size, carrier gas flow rate, column properties, and temperature affect the separation. GLC allows both qualitative and quantitative analysis of many organic compounds.
Chromatography is a technique used to separate and identify the components of a mixture. It works by allowing molecules to distribute themselves between a stationary and mobile phase, so that molecules that interact more with the mobile phase move faster. Chromatographic techniques can be classified based on the interaction with the stationary phase or physical state of the mobile phase. Key techniques include adsorption, partition, ion exchange, exclusion, gas, liquid, and thin layer chromatography. Proper sample preparation and development conditions are important for achieving optimal separation and resolution of components in the mixture.
Chromatography is a scientific technique used to separate mixtures based on how compounds interact with two phases - a stationary phase and a mobile phase. There are several types of chromatography including thin layer chromatography, gas chromatography, high performance liquid chromatography, electrophoresis, and paper chromatography. Each type uses different stationary and mobile phases and has various applications such as determining compound compositions, analyzing organic reactions, and identifying unknown substances.
The document discusses the process and history of paper chromatography. It begins with an introduction to chromatography and its use in separating mixtures. It then covers the history from early experiments in the 1860s to modern developments. The main types and techniques of paper chromatography are explained, including the use of a stationary phase, mobile phase, and capillary action to separate components by affinity and travel distance. Key steps like sample application and developing the paper strip are outlined.
The document describes planar chromatography techniques, specifically thin layer chromatography (TLC). It explains that TLC separates mixtures by using a thin stationary phase like silica gel coated on a plate and a mobile phase liquid solvent. The steps of TLC are described as sample application, development where separation occurs, visualization under UV light, and interpretation by calculating Rf values. Applications for separating lipids, carbohydrates and other compounds are outlined.
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 technique used to separate mixtures by distributing components between a stationary and mobile phase. It works on the principle that different compounds interact differently with the phases and therefore move through the system at different rates. There are various types of chromatography classified by mobile phase (gas or liquid) or interaction forces (adsorption, partition, ion exchange). Key components are the mobile phase, stationary phase, and supporting medium. Chromatography is widely used in fields like analytical chemistry, biochemistry, environmental analysis and forensic science.
This document discusses different types of chromatography techniques including paper chromatography, gas chromatography, thin layer chromatography, and high performance thin layer chromatography. It provides details on the principles, processes, and applications of each technique. Paper chromatography separates colored chemicals or substances based on partition between a mobile phase that moves over stationary filter paper. Gas chromatography uses an inert gas as the mobile phase to separate compounds that can be vaporized. Thin layer chromatography separates compounds on a thin layer of absorbent material coated on a plate based on differing affinities to the stationary phase. High performance thin layer chromatography is an enhanced form of thin layer chromatography that can automate steps and provide more accurate quantitative measurements. Each technique has various applications in fields like pharmaceutical analysis,
Chromatography is a technique used to separate and identify components of a mixture. It works by distributing molecules between a stationary and mobile phase. Molecules that spend more time in the mobile phase move faster through the column. There are several types of chromatography classified by mobile phase or separation mechanism, including gas chromatography which uses gases, thin layer chromatography which uses adsorbents on plates, and liquid chromatography which uses liquids. Chromatography is used in various applications such as pharmaceutical analysis, environmental monitoring, and forensic analysis.
High Performance Liquid chromatography (HPLC)Unnati Garg
The document provides an overview of high performance liquid chromatography (HPLC). It describes key components of HPLC systems including pumps to deliver solvent at stable flow rates, columns for molecular separation, and detectors for recognizing analytes. The separation principle is based on the distribution of analytes between a mobile liquid phase and stationary column packing material. Different constituents are eluted at different times, achieving separation. HPLC is widely used in pharmaceutical applications such as drug development, production quality control, and stability testing.
Chromatography is a method of separating components of a mixture through their interactions with two phases - a stationary phase and a mobile phase. The components are distributed between the phases based on properties like solubility and affinity. There are several types of chromatography classified by the shape of the stationary phase (e.g. thin layer), the state of the mobile phase (e.g. gas, liquid), or the interaction between solute and stationary phase (e.g. adsorption, partition). Chromatography techniques are used in various applications including pharmaceutical quality control, forensic analysis, and biological research like protein purification.
Chromatography is a technique used to separate mixtures into individual components. Paper chromatography is a type of chromatography that uses paper as the stationary phase. The mixture is applied to the paper and then placed in a developing chamber with the mobile phase solvent. As the solvent travels up the paper, the different components of the mixture separate based on how strongly they interact with the stationary and mobile phases. This creates discrete spots that can be analyzed to identify the components in the original mixture. Paper chromatography is a simple, inexpensive, and effective technique for separating and analyzing mixtures.
Thin layer chromatography (TLC) is a technique used to separate mixtures into their components. It involves a stationary phase, such as a silica gel-coated plate, and a mobile phase, which can be a solvent or solvent mixture. Samples are spotted onto the plate and the mobile phase is allowed to travel up the plate, separating the samples based on how strongly they interact with each phase. TLC provides a simple, low-cost method for analyzing mixtures and is useful in fields like pharmaceuticals, biochemistry, and food and cosmetic analysis.
Chromatography is a technique used to separate the components of a mixture through differential partitioning between a stationary and mobile phase. There are various types of chromatography classified by the physical state of the phases used and the separation mechanism employed. The document discusses the basic principles and history of chromatography. It describes different techniques like paper chromatography, thin layer chromatography, gas chromatography, liquid chromatography and ion exchange chromatography. Applications and significance of these techniques in fields like pharmaceuticals, forensics and food analysis are also highlighted.
The document discusses the principles of chromatography. It describes how chromatography separates components in a mixture based on differences in their interactions with mobile and stationary phases. It discusses how Michael Tswett first demonstrated chromatography in 1903 and the key aspects of how it works. These include how retention time, partition coefficients, selectivity factors and efficiency parameters like plate number and height equivalent to a theoretical plate are used to characterize chromatographic separations.
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 overview of chromatography techniques. It discusses the definition and history of chromatography, and describes several types including paper chromatography, thin layer chromatography, column chromatography, gas chromatography, and high performance liquid chromatography. Specific details are provided on the principles, procedures, applications, advantages and disadvantages of paper chromatography and thin layer chromatography. Key differences between these two techniques are also compared.
What is Chromatography?
Applications of Chromatography
Types of Chromatography
1- Column Chromatography
2- Planar chromatography
Paper Chromatography
Gas Chromatography
Detectors
Reverse phase chromatography is a technique where the binding of solutes to a hydrophobic stationary phase occurs via hydrophobic interactions. It uses a stationary phase with hydrophobic ligands chemically bonded to a solid support. Polar mobile phases are used to elute retained solutes from the column. Key parameters that affect separation include the pH, ionic strength, and polarity of the mobile phase, use of gradients or isocratic elution, column length, and addition of ion-pairing agents. Reverse phase chromatography is commonly used to purify biomolecules like proteins, peptides, and nucleic acids.
Partition Chromatography technique is defined as. the separation of components between two liquid phases viz original solvent and the film of solvent used in the column.
The document discusses column chromatography. It begins with an introduction to chromatography, including its history and definitions. It then covers various types of chromatography and the principles and requirements of column chromatography. The key factors that affect column chromatography are the stationary and mobile phases used. Various stationary phases are described, including silica gel and adsorbents. The document also discusses preparation of the column and factors that influence column efficiency.
Chromatography is a technique used to separate mixtures and involves dissolving the mixture in a mobile phase that carries it through a structure containing a stationary phase. It was first developed in 1900 by Russian scientist Mikhail Tsvet and involves the interaction between a mobile and stationary phase resulting in separation. Common types include paper chromatography, thin layer chromatography, gas chromatography, and affinity chromatography. It has various applications in fields like clinical analysis, protein purification, forensic toxicology, and separation of amino acids, proteins, carbohydrates, and more.
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.
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.
Water activity is a measure of how available water is in a food system for microbial growth. It is defined as the vapor pressure of water in a food divided by the vapor pressure of pure water at the same temperature. Measurement techniques include vapor pressure measurement using a manometer, freezing point depression, psychrometry using hygrometers or thermocouples, isopiestic transfer between materials, and measuring matric or suction potential in gels. Choice of technique depends on required range, accuracy, precision and speed needed.
Chromatography is a technique used to separate mixtures into individual components and analyze, identify, and purify substances. It works by distributing components between a stationary and mobile phase as they travel through a chromatographic system. The visual output, or chromatogram, shows different patterns corresponding to different mixture components based on their retention times and concentrations. Common stationary phases include adsorbents like liquids or solids, while mobile phases are usually solvents or supercritical fluids that carry analytes through the system.
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 technique used to separate mixtures by distributing components between a stationary and mobile phase. It works on the principle that different compounds interact differently with the phases and therefore move through the system at different rates. There are various types of chromatography classified by mobile phase (gas or liquid) or interaction forces (adsorption, partition, ion exchange). Key components are the mobile phase, stationary phase, and supporting medium. Chromatography is widely used in fields like analytical chemistry, biochemistry, environmental analysis and forensic science.
This document discusses different types of chromatography techniques including paper chromatography, gas chromatography, thin layer chromatography, and high performance thin layer chromatography. It provides details on the principles, processes, and applications of each technique. Paper chromatography separates colored chemicals or substances based on partition between a mobile phase that moves over stationary filter paper. Gas chromatography uses an inert gas as the mobile phase to separate compounds that can be vaporized. Thin layer chromatography separates compounds on a thin layer of absorbent material coated on a plate based on differing affinities to the stationary phase. High performance thin layer chromatography is an enhanced form of thin layer chromatography that can automate steps and provide more accurate quantitative measurements. Each technique has various applications in fields like pharmaceutical analysis,
Chromatography is a technique used to separate and identify components of a mixture. It works by distributing molecules between a stationary and mobile phase. Molecules that spend more time in the mobile phase move faster through the column. There are several types of chromatography classified by mobile phase or separation mechanism, including gas chromatography which uses gases, thin layer chromatography which uses adsorbents on plates, and liquid chromatography which uses liquids. Chromatography is used in various applications such as pharmaceutical analysis, environmental monitoring, and forensic analysis.
High Performance Liquid chromatography (HPLC)Unnati Garg
The document provides an overview of high performance liquid chromatography (HPLC). It describes key components of HPLC systems including pumps to deliver solvent at stable flow rates, columns for molecular separation, and detectors for recognizing analytes. The separation principle is based on the distribution of analytes between a mobile liquid phase and stationary column packing material. Different constituents are eluted at different times, achieving separation. HPLC is widely used in pharmaceutical applications such as drug development, production quality control, and stability testing.
Chromatography is a method of separating components of a mixture through their interactions with two phases - a stationary phase and a mobile phase. The components are distributed between the phases based on properties like solubility and affinity. There are several types of chromatography classified by the shape of the stationary phase (e.g. thin layer), the state of the mobile phase (e.g. gas, liquid), or the interaction between solute and stationary phase (e.g. adsorption, partition). Chromatography techniques are used in various applications including pharmaceutical quality control, forensic analysis, and biological research like protein purification.
Chromatography is a technique used to separate mixtures into individual components. Paper chromatography is a type of chromatography that uses paper as the stationary phase. The mixture is applied to the paper and then placed in a developing chamber with the mobile phase solvent. As the solvent travels up the paper, the different components of the mixture separate based on how strongly they interact with the stationary and mobile phases. This creates discrete spots that can be analyzed to identify the components in the original mixture. Paper chromatography is a simple, inexpensive, and effective technique for separating and analyzing mixtures.
Thin layer chromatography (TLC) is a technique used to separate mixtures into their components. It involves a stationary phase, such as a silica gel-coated plate, and a mobile phase, which can be a solvent or solvent mixture. Samples are spotted onto the plate and the mobile phase is allowed to travel up the plate, separating the samples based on how strongly they interact with each phase. TLC provides a simple, low-cost method for analyzing mixtures and is useful in fields like pharmaceuticals, biochemistry, and food and cosmetic analysis.
Chromatography is a technique used to separate the components of a mixture through differential partitioning between a stationary and mobile phase. There are various types of chromatography classified by the physical state of the phases used and the separation mechanism employed. The document discusses the basic principles and history of chromatography. It describes different techniques like paper chromatography, thin layer chromatography, gas chromatography, liquid chromatography and ion exchange chromatography. Applications and significance of these techniques in fields like pharmaceuticals, forensics and food analysis are also highlighted.
The document discusses the principles of chromatography. It describes how chromatography separates components in a mixture based on differences in their interactions with mobile and stationary phases. It discusses how Michael Tswett first demonstrated chromatography in 1903 and the key aspects of how it works. These include how retention time, partition coefficients, selectivity factors and efficiency parameters like plate number and height equivalent to a theoretical plate are used to characterize chromatographic separations.
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 overview of chromatography techniques. It discusses the definition and history of chromatography, and describes several types including paper chromatography, thin layer chromatography, column chromatography, gas chromatography, and high performance liquid chromatography. Specific details are provided on the principles, procedures, applications, advantages and disadvantages of paper chromatography and thin layer chromatography. Key differences between these two techniques are also compared.
What is Chromatography?
Applications of Chromatography
Types of Chromatography
1- Column Chromatography
2- Planar chromatography
Paper Chromatography
Gas Chromatography
Detectors
Reverse phase chromatography is a technique where the binding of solutes to a hydrophobic stationary phase occurs via hydrophobic interactions. It uses a stationary phase with hydrophobic ligands chemically bonded to a solid support. Polar mobile phases are used to elute retained solutes from the column. Key parameters that affect separation include the pH, ionic strength, and polarity of the mobile phase, use of gradients or isocratic elution, column length, and addition of ion-pairing agents. Reverse phase chromatography is commonly used to purify biomolecules like proteins, peptides, and nucleic acids.
Partition Chromatography technique is defined as. the separation of components between two liquid phases viz original solvent and the film of solvent used in the column.
The document discusses column chromatography. It begins with an introduction to chromatography, including its history and definitions. It then covers various types of chromatography and the principles and requirements of column chromatography. The key factors that affect column chromatography are the stationary and mobile phases used. Various stationary phases are described, including silica gel and adsorbents. The document also discusses preparation of the column and factors that influence column efficiency.
Chromatography is a technique used to separate mixtures and involves dissolving the mixture in a mobile phase that carries it through a structure containing a stationary phase. It was first developed in 1900 by Russian scientist Mikhail Tsvet and involves the interaction between a mobile and stationary phase resulting in separation. Common types include paper chromatography, thin layer chromatography, gas chromatography, and affinity chromatography. It has various applications in fields like clinical analysis, protein purification, forensic toxicology, and separation of amino acids, proteins, carbohydrates, and more.
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.
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.
Water activity is a measure of how available water is in a food system for microbial growth. It is defined as the vapor pressure of water in a food divided by the vapor pressure of pure water at the same temperature. Measurement techniques include vapor pressure measurement using a manometer, freezing point depression, psychrometry using hygrometers or thermocouples, isopiestic transfer between materials, and measuring matric or suction potential in gels. Choice of technique depends on required range, accuracy, precision and speed needed.
Chromatography is a technique used to separate mixtures into individual components and analyze, identify, and purify substances. It works by distributing components between a stationary and mobile phase as they travel through a chromatographic system. The visual output, or chromatogram, shows different patterns corresponding to different mixture components based on their retention times and concentrations. Common stationary phases include adsorbents like liquids or solids, while mobile phases are usually solvents or supercritical fluids that carry analytes through the system.
This document provides an introduction to quantitative structure-activity relationships (QSAR). It defines QSAR as quantifying physicochemical properties of drugs to see their effect on biological activity. Graphs are used to plot activity versus properties, and regression analysis determines correlation. Key properties discussed are hydrophobicity, steric effects, and electronic effects. Hansch analysis uses equations to relate activity to multiple properties. Advantages are understanding structure-activity and enabling novel analog design. Limitations include potential for false correlations and need for large, high-quality datasets.
QSAR attempts to quantify physicochemical properties like hydrophobicity, electronics, and steric effects and relate them to biological activity. Log P measures hydrophobicity and is often plotted against biological activity, sometimes giving a straight line and other times a parabolic curve. Other parameters like π, σ, and Es measure substituent hydrophobicity, electronic effects, and steric effects. QSAR equations combine these parameters to model and predict biological activity for a series of compounds.
The document describes the development and refinement of a quantitative structure-activity relationship (QSAR) model to predict the biological activity of pyranenamine compounds. It discusses 5 stages of synthesizing analogs and developing the QSAR equation based on substituents. Anomalies identified were used to refine the model terms. The final optimized QSAR equation considered parameters like hydrophilicity, hydrogen bonding, resonance effects, and steric hindrance to identify a hypothetical compound over 1000 times more active than the lead compound.
Liquid-liquid extraction is a separation process where one or more components of a liquid mixture are preferentially dissolved into another immiscible liquid solvent based on factors like distribution coefficient, selectivity, density, and chemical reactivity. Solvent choice depends on these factors as well as properties like insolubility, recoverability, viscosity, vapor pressure, freezing point, availability, and cost. Extraction is often preferred over distillation when relative volatility is near unity, azeotropes would limit separation, heating must be avoided, or components are very different in nature.
Quantitative structure-activity relationships (QSAR) use mathematical models to predict biological activity based on molecular properties. QSAR models are developed using statistical methods like partial least squares on datasets of compounds with known activities. Three-dimensional (3D) QSAR extends this approach by incorporating 3D structural descriptors and molecular fields derived from programs like CoMFA, VolSurf, and Catalyst to model activity based on interactions at binding sites. These 3D-QSAR models can be used to predict activity and design new compounds with improved properties.
The document discusses specifications for raw materials, finished products, and documentation. It defines specifications and their importance in ensuring quality. Specifications include tests, acceptance criteria, and procedures for materials and products. The document outlines specifications for raw material purchasing, storage, and testing as well as finished product testing, documentation, and storage. It discusses the relationship between specifications and regulations/standards.
This document provides background information on thin-layer chromatography (TLC) and column chromatography. It describes the basic two-part operation involving a mobile phase that carries samples through a stationary phase. The document discusses factors that influence separation, such as choosing adsorbents and eluents. It then describes how to perform TLC and column chromatography experiments, including preparing plates/columns, developing separations, and analyzing results. Procedures are outlined for identifying unknown compounds and disposing of waste.
The document discusses different types of solid forms that active pharmaceutical ingredients can take, including polymorphs, solvates, hydrates, salts, co-crystals, and the amorphous form. It notes that over 80% of pharmaceutical solids exhibit polymorphism. The thermodynamically most stable polymorph is generally preferred for stability reasons, though a metastable polymorph may be developed to provide a balance between processability and stability. Hydrates and solvates are discussed, with hydrates being the most common type of solvate. Salt and co-crystal formation can impact properties like dissolution and stability. The amorphous form lacks long-range order.
The document discusses Quantitative Structure Activity Relationships (QSAR), which attempt to identify and quantify physicochemical properties of drugs that influence biological activity using mathematical equations. It describes key physicochemical properties considered in QSAR like hydrophobicity, electronic effects, and steric effects. Measurement scales for these properties like log P, π, σ, and Es are defined. The Hansch equation is presented as a typical QSAR model relating these factors to biological activity. Craig plots are also introduced to help select substituents for QSAR analysis.
This document provides instructions for separating acids and neutral compounds using solvent extraction. It defines extraction and washing, and discusses choosing appropriate solvents based on polarity. For acid-base extractions, a weaker acid will dissolve in water while a stronger acid will dissolve in an organic solvent. The experimental procedure uses these principles to separate a mixture of p-toluic acid, p-tert-butylphenol, and acetanilide through a series of extractions with sodium bicarbonate solution, sodium hydroxide solution, and organic solvents. Key steps include isolating p-toluic acid through acidification, and isolating p-tert-butylphenol through heating, cooling
This document describes an experiment to separate a mixture of three organic compounds - cinnamic acid, p-toluidine, and anisole - using acid/base extraction with dichloromethane. The compounds are separated using liquid-liquid extraction with hydrochloric acid and sodium hydroxide to isolate the acidic, basic, and neutral compounds into different layers. 1H NMR spectra are taken during the separation process and of the purified compounds to analyze the effectiveness of the separation technique. Percent yields are also calculated for each separated compound.
This document provides an overview of quantitative structure-activity relationship (QSAR) modeling techniques. It discusses:
1) The history and background of QSAR, dating back to the 19th century, and key contributors like Hammett who developed linear free energy relationships.
2) Common QSAR methodologies like multiple linear regression, principal component analysis, partial least squares, artificial neural networks, and genetic algorithm-based approaches.
3) Steps for validating QSAR models, including correlation coefficients, cross-validation, and assessing the applicability domain for making predictions.
A QSAR is a mathematical relationship between a biological activity of a molecular system and its geometric and chemical characteristics.
QSAR attempts to find consistent relationship between biological activity and molecular properties, so that these “rules” can be used to evaluate the activity of new compounds.
QSAR attempts to correlate biological activity to measurable physicochemical properties through mathematical equations. It relates parameters like lipophilicity (log P), electronic effects (Hammett constants), and steric effects to biological response. Various QSAR methods exist, including Hansch analysis which uses these substituent constants in its equations. More advanced techniques like CoMFA analyze fields around aligned molecules to model activity landscapes and identify favorable regions for activity. QSAR provides a framework for drug design and predicting activities of untested compounds.
The document discusses the structure-activity relationships of various antipsychotic drugs. It begins by describing the dopamine receptor and how drugs like phenothiazines are able to bind to it. It then covers specific structural features of different drug classes that impact their potency and side effect profiles, including phenothiazines, butyrophenones, and atypical antipsychotics. Key structural attributes discussed are the presence of electron-withdrawing groups on phenothiazine rings, different side chains on phenothiazines and butyrophenones, and the chemical structures of atypical antipsychotics. The document concludes that while older structure-function concepts were important, current antipsychotics have diverse mechanisms of action involving
The document presents a research project on preparing and evaluating co-crystals of atorvastatin calcium with saccharin and urea. The objectives are to prepare the co-crystals using solvent drop grinding and solvent evaporation methods, characterize them using various techniques, and evaluate their solubility. The document outlines the materials, methods, results and discussion sections of the project including pre-formulation studies of atorvastatin calcium and the co-formers, characterization of the prepared co-crystals using FTIR, DSC and solubility testing, and evaluation of drug-excipient compatibility. The project aims to develop co-crystal formulations to improve the solubility and dissolution rate of the poorly soluble at
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with stationary and mobile phases. There are several types that differ based on the phases used, including paper, column, thin layer, gas, and liquid chromatography. Chromatography was developed in the early 20th century and has since been used across various fields like forensics and environmental testing to separate and analyze sample components.
Chromatography is a laboratory technique used to separate mixtures by exploiting differences in how components partition between a stationary and mobile phase. The document discusses various chromatography techniques including column chromatography, planar chromatography like paper and thin layer chromatography, displacement chromatography, gas chromatography using a gas mobile phase, liquid chromatography using a liquid mobile phase like HPLC, and affinity chromatography which uses specific non-covalent interactions for purification. Key terms related to chromatography components and processes are also defined.
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.
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 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
Chromatography is an analytical technique used to separate mixtures based on differences in physical properties of constituents. It involves a stationary phase and mobile phase. In chromatography, mixtures are separated as they travel through the stationary phase at different rates depending on interactions with the phases. Common types include paper chromatography, thin layer chromatography, column chromatography, and high performance liquid chromatography which uses high pressure to enhance separation.
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 by narayan sarkar and simi baruah new versionNarayanSarkar6
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with two phases - a stationary phase and a mobile phase. The document provides an introduction to chromatography, including its history, principles, types (column, paper, thin layer, affinity), and applications. Column chromatography involves passing a mixture through a column containing a stationary phase to separate components based on differences in how strongly they adhere to the stationary phase.
This document provides an overview of chromatography. It begins with an introduction that defines chromatography and describes how it separates mixtures based on differences in solubility between components in mobile and stationary phases. The document then covers the history of chromatography, important technical terms, main types categorized by interaction with the stationary phase or physical state of the mobile phase. Applications are discussed in areas like drug development, food testing, and forensics. Advantages are noted as versatility in separation. Disadvantages include temperature sensitivity and ensuring solubility. References are listed at the end.
This document provides an introduction to analytical separation techniques and chromatography. It discusses classical and instrumental methods of analysis, with instrumental methods using physical properties and efficient separation techniques. Chromatography is introduced as a physical method that separates analytes distributed between two phases, one stationary and one mobile. Key terms like mobile phase, stationary phase, and supporting medium are defined. Different types of chromatography are classified based on the physical means of separation, type of mobile/stationary phases, and type of interaction between analyte and stationary phase. Important chromatography concepts like elution, resolution, migration rates, distribution constants, and theoretical plates are also introduced.
Chromatography is a laboratory technique used to separate mixtures of compounds into their individual components based on how they interact with different substances. It works by taking advantage of differences in how molecules move through stationary and mobile phases. The key principles are that separation is achieved as compounds differentially interact and partition between phases based on properties like polarity, size, and solubility. Chromatography has many applications and is widely used in fields like industrial analysis, medicine, food testing, and environmental science.
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with stationary and mobile phases. It involves passing a mixture through a stationary phase that separates components based on properties like charge, solubility, or adsorption. Main types include liquid chromatography, paper chromatography, gas chromatography, and thin layer chromatography. Chromatography has advanced separation capabilities and is used in applications like analyzing pollution, forensic analysis, and purifying compounds.
Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with two phases - a stationary phase and a mobile phase. There are several types of chromatography including paper chromatography, thin layer chromatography, column chromatography, and ion exchange chromatography. Each type uses a different stationary phase and separation method to isolate analytes in a mixture.
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.
Chromatography is an analytical technique used to separate mixtures based on differences in physical properties of constituents. It involves a stationary phase and mobile phase. In chromatography, mixtures are separated as they travel through the stationary phase at different rates depending on interactions with the phases. Common types include paper chromatography, thin layer chromatography, column chromatography, and high performance liquid chromatography which uses high pressure to enhance separation.
This document discusses the history and types of chromatography. It begins by explaining that chromatography derives from Greek words meaning "written in color" and was developed in the early 20th century by Russian botanist Michail Semenovich Tswett. The document then describes the basic principles of chromatography and lists the main types: adsorption, partition, ion exchange, exclusion, and affinity chromatography. It provides details on each type and concludes by discussing different stationary and mobile phases used in chromatography.
This document provides an introduction and overview of chromatography techniques. It discusses the basic setup and history of chromatography. It also classifies different types of chromatography based on phases and shape of the chromatographic bed. The document specifically describes adsorption and partition column chromatography, including their theory, terminology, methodology and applications. Adsorption chromatography utilizes a mobile phase that is adsorbed onto a stationary solid phase, while partition chromatography uses a thin liquid film stationary phase.
Chromatography is a technique used to separate components of mixtures. It works by passing a sample mixture through a stationary phase as a mobile phase carries the components through at different rates based on interactions with the stationary phase. This allows the components to be separated into individual fractions. There are several types of chromatography that differ based on the stationary and mobile phases used, including gas chromatography, liquid chromatography, and thin layer chromatography. Chromatography has many applications in fields like pharmaceuticals, chemicals, foods, forensics, and molecular biology.
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.
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Chromatography
1. Chromatography
Chromatography is the term used to describe a separation technique in which a mobile
phase carrying a mixture is caused to move in contact with a selectively absorbent stationary
phase. There are number of different kinds of chromatography, which differ in the mobile
and the stationary phase used.
Chromatography is derived from a Greek word χρῶμα means chroma (colour) and γράφειν
means graphein (to write). Chromatography is the laboratory techniques used 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. Subtle differences in a
compound's partition coefficient result in differential retention on the stationary phase and
thus changing the separation.
Chromatography may be preparative or analytical. The purpose of preparative
chromatography is to separate the components of a mixture for more advanced use (and is
thus a form of purification). Analytical chromatography is done normally with smaller
amounts of material and is for measuring the relative proportions of analytes in a mixture.
The two are not mutually exclusive.
2. History of chromatography.......
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 colours (green, orange, and yellow,
respectively) they gave the technique its name. New types of chromatography developed
during the 1930s and 1940s made the technique useful for many separation processes.
Chromatography technique developed substantially as a result of the work of Archer John
Porter Martin and Richard Laurence Millington Synge during the 1940s and 1950s. They
established the principles and basic techniques of partition chromatography, and their work
encouraged the rapid development of several chromatographic methods: Paper
Chromatography, Gas Chromatography, and what would become known as high
performance liquid chromatography. Since then, the technology has advanced rapidly.
Researchers found that the main principles of Tsvet's chromatography could be applied in
many different ways, resulting in the different varieties of chromatography described below.
Advances are continually improving the technical performance of chromatography, allowing
the separation of increasingly similar molecules.
Techniques by chromatographic bed shape.......
1. paper chromatography
In Paper Chromatography, the mobile phase is a
solvent and the stationary phase is water held in the
fibres of chromatography paper. A solution of the
mixture to be separated is spotted onto a strip of
chromatography paper (or filter paper) with a dropper.
The chromatogram is developed by placing the bottom
of the paper (but not the sample spot) in a tank
containing suitable solvent. The solvent is drawn up the
paper by capillary action. The components of the mixture move up the paper with the solvent
at different rates due to their differing interactions with the stationary and mobile phases.
Formula-
Rf = Distance the solute moves
Distance the solvent front move
3. 2. Column chromatography
In column chromatography, the mobile phase
is again a solvent, and the stationary phase is a
finely divided solid, such as silica gel or
alumina. Chromatography columns vary in size
and polarity. There is an element of trial and
error involved in selecting a suitable solvent
and column for the separation of the
constituents of a particular mixture. A small
volume of the sample whose constituents are to
be separated is placed on top of the column.
The choice of the eluting solvent should ensure
that the sample is soluble. However, if the
sample was too soluble the mobile phase
(solvent) would move the solutes too quickly,
resulting in the non-separation of the different
constituents.
3. Thin layer chromatography
In thin layer chromatography, the
mobile phase is also a solvent, and
the stationary phase is a thin layer
of finely divided solid, such as
silica gel or alumina, supported on
glass or aluminium. Thin layer
chromatography is similar to paper
chromatography in that it involves
spotting the mixture on the plate
and the solvent (mobile phase) rises up the plate in the chromatography
tank. It has an advantage over paper chromatography in that its
4. separations are very efficient because of the much smaller size of the
particles in the stationary phase.
Thin layer chromatography is particularly useful in forensic work, for example
in the separation of dyes from fibres. Gas chromatography and high
performance liquid chromatography are more sophisticated chromatographic
techniques.
4. Gas chromatography
A gas is the mobile phase and the stationary phase can be either a solid or a
non- volatile liquid.
There are five basic GC components:
1) Pneumatic system – gas supply (flow control and measurement).
2) Injection system – a heated injector port, where the sample is vaporised
if necessary.
3) Column – where the separation occurs.
4) Oven –The coiled column is wholly contained in a thermostatically
controlled oven.
5) Detector – integral detector or link to a mass spectrometer.
How does gas chromatography work.........?
1) A carrier gas, examples of which are Helium and Neon flows through the
system. A valve controls the flow rate.
2) A sample of the volatile mixture is injected into the carrier gas. The
sample is vaporised in the heated injector port.
5. 3) The carrier gas carries the vaporised sample into the column. The columns
are stainless steel or glass tubes. They can be up to 25 m in length and are
of narrow bore (2-10 mm). Therefore the column is often wound into a
coil. The packed columns contain porous support material. The sample
mixture undergoes a series of interactions between the stationary and
mobile phases as it is carried through the system by the carrier gas. Due
to the wide choice of materials available for the stationary and mobile
phases, it is possible to separate molecules that differ only slightly in their
physical and chemical properties.
4) The coiled column is contained in the thermostatically controlled oven.
5) Separated components emerge in the order of increasing interaction with
the stationary phase. The least retarded component comes through first.
Separation is obtained when one compound is sufficiently retarded to
prevent overlap with another component of the sample, as it emerges
from the column.
6) Two types of detector can be used:
(1) Thermal Conductivity detectors which respond to changes in the thermal
conductivity of the gas leaving the column and
(2) Flame Ionisation detection (FID), which is more commonly used. In
thermal conductivity, as the carrier gas leaves the column, it cools the
detector. When a solute emerges with the carrier gas, it does not cool the
detector to the same extent. Alternatively, samples can be passed from the
oven directly into a mass spectrometer, where they are analysed.
Retention time is defined as the time taken for a component to go from injection
to detection. This varies depending on
a) The nature of and the interactions between the solute and the stationary
and mobile phases.
b) The flow rate of the carrier gas,
c) The temperature of the column (shorter retention times are obtained at
higher temperatures),
d) The length and diameter of the column,
6. Once GC has separated a mixture, the components can be identified using
known retention times. For unknown compounds the solutes are collected
individually and analysed using another method, e.g. mass spectrometry.
For each compound in a mixture one peak is observed on the chromatogram. In
the particular set of operating conditions relating to the column, the retention
time will increase with the size and polarity of the compound. To find the
concentration of a particular compound, the peak height should be measured.
GC is used to analyse blood samples for the presence of alcohol. It is also used
to analyse samples taken from athletes to check for the presence of drugs. In
each case, it separates the components of the mixture and indicates the
concentrations of the components. Water companies test samples of water for
pollutants using GC to separate the pollutants, and mass spectrometry to
identify them.
GC is used to analyse blood samples for the presence of alcohol. It is also used
to analyse samples taken from athletes to check for the presence of drugs. In
each case, it separates the components of the mixture and indicates the
concentrations of the components. Water companies test samples of water for
pollutants using GC to separate the pollutants, and mass spectrometry to
identify them.
7. 5. High performance liquid chromatography
Basic Components:
1) Solvent Reservoir.
2) The Pump System controls the flow and measures the volume of solvent
(the mobile phase). The flow rates of HPLC columns are slow – often in
3 -1
the range of 0.5 - 5 cm min .
3) The Injector System: The sample to be separated is injected into the
liquid phase at this point.
4) The Column is made of steel and packed usually with porous silica
particles (the stationary phase). Different materials can be used depending
on the nature of the liquid. A long column is not needed because
separation in HPLC is very efficient. Columns are usually 10 –30 cm
long, with an internal diameter of 4 mm.
Different components of the sample are carried forward at different
rates by the moving liquid phase, due to their differing interactions
with the stationary and mobile phases.
5) The Detector: When the components reach the end of the column they
are analysed by a detector. The amounts passing through the column are
small, so solutes are analysed as they leave the column. Therefore HPLC
is usually linked to a spectrometer (e.g. ultra violet or mass
spectrometry).
The length of time it takes for a compound to reach the detector allows
the component to be identified. Like the GC, once the retention time
of a solute has been established for a column using a particular set of
operating conditions, the solute can be identified in a mixture. A
chromatogram is obtained for the sample.
8. Uses
HPLC has many uses such as drug testing, testing for vitamins in food and
growth promoters in meat. In each case components of the mixture are
separated and detected.
Comparison of HPLC over GC
Less volatile and larger samples can be used with HPLC.
9. Chromatogram and Mass Spectrometry Data
6. Affinity chromatography
Affinity chromatography is based on selective non-covalent
interaction between an analyte and specific molecules. It is very
specific, but not very robust. It is often used in biochemistry in the
purification of proteins bound to tags. These fusion proteins are
labelled with compounds such as his-tags, biotin or antigens, which
bind to the stationary phase specifically. After purification, some of
these tags are usually removed and the pure protein is obtained.
Affinity chromatography often utilizes a bio-molecule's affinity for a
metal (Zn, Cu, Fe, etc.). Columns are often manually prepared.
Traditional affinity columns are used as a preparative step to flush out
unwanted bio-molecules. However, HPLC techniques exist that do
utilize affinity chromatography properties. Immobilized Metal
Affinity Chromatography (IMAC) is useful to separate
aforementioned molecules based on the relative affinity for the metal
(i.e. Dionex IMAC). Often these columns can be loaded with different
metals to create a column with a targeted affinity.
10. 7. Super critical fluid chromatography
Supercritical fluid chromatography is a separation technique in which the
mobile phase is a fluid above and relatively close to its critical temperature and
pressure.
Techniques by separation mechanism..........
8. Ion exchange chromatography
Ion exchange chromatography (usually referred to as ion chromatography) uses
an ion exchange mechanism to separate analytes based on their respective
charges. It is usually performed in columns but can also be useful in planar
mode. Ion exchange chromatography uses a charged stationary phase to
separate charged compounds including anions, cations, amino acids, peptides,
and proteins. In conventional methods the stationary phase is an ion exchange
resin that carries charged functional groups that interact with oppositely charged
groups of the compound to retain. Ion exchange chromatography is commonly
used to purify proteins using FPLC.
9. Size exclusion chromatography
Size-exclusion chromatography (SEC) is also known as Gel Permeation
Chromatography (GPC) or Gel Filtration Fhromatography and separates
molecules according to their size (or more accurately according to their
hydrodynamic diameter or hydrodynamic volume). Smaller molecules are able
to enter the pores of the media and, therefore, molecules are trapped and
removed from the flow of the mobile phase. The average residence time in the
pores depends upon the effective size of the analyte molecules. However,
molecules that are larger than the average pore size of the packing are excluded
and thus suffer essentially no retention; such species are the first to be eluted. It
is generally a low-resolution chromatography technique and thus it is often
reserved for the final, "polishing" step of a purification. It is also useful for
determining the tertiary structure and quaternary stucture of purified proteins,
especially since it can be carried out under native solution conditions.
11. Special techniques..........
10. reversed phase chromatography
Reversed-phase chromatography is an elution procedure used in liquid
chromatography in which the mobile phase is significantly more polar than the
stationary phase.
11. two-dimensional chromatography
In some cases, the chemistry within a given column can be insufficient to
separate some analytes. It is possible to direct a series of unresolved peaks onto
a second column with different physico-chemical (Chemical classification)
properties. Since the mechanism of retention on this new solid support is
different from the first dimensional separation, it can be possible to separate
compounds that are indistinguishable by one-dimensional chromatography. The
sample is spotted at one corner of a square plate, developed, air-dried, then
rotated by 90° and usually redeveloped in a second solvent system.
12. pyrolysis gas chromatography
Pyrolysis gas chromatography mass spectrometry is a method of chemical
analysis in which the sample is heated to decomposition to produce smaller
molecules that are separated by gas chromatography and detected using mass
spectrometry.
Pyrolysis is the thermal decomposition of materials in an inert atmosphere or a
vacuum. The sample is put into direct contact with a platinum wire, or placed in
a quartz sample tube, and rapidly heated to 600–1000 °C. Depending on the
application even higher temperatures are used. Three different heating
techniques are used in actual pyrolyzers: Isothermal furnace, inductive heating
(Curie Point filament), and resistive heating using platinum filaments. Large
molecules cleave at their weakest points and produce smaller, more volatile
fragments. These fragments can be separated by gas chromatography. Pyrolysis
GC chromatograms are typically complex because a wide range of different
decomposition products is formed. The data can either be used as fingerprint to
12. prove material identity or the GC/MS data is used to identify individual
fragments to obtain structural information. To increase the volatility of polar
fragments, various methylating reagents can be added to a sample before
pyrolysis.
Besides the usage of dedicated pyrolyzers, pyrolysis GC of solid and liquid
samples can be performed directly inside Programmable Temperature Vaporizer
(PTV) injectors that provide quick heating (up to 30 °C/s) and high maximum
temperatures of 600–650 °C. This is sufficient for some pyrolysis applications.
The main advantage is that no dedicated instrument has to be purchased and
pyrolysis can be performed as part of routine GC analysis. In this case quartz
GC inlet liners have to be used. Quantitative data can be acquired, and good
results of derivatization inside the PTV injector are published as well.
13. fast protein liquid chromatography
Fast protein liquid chromatography (FPLC) is a term applied to several
chromatography techniques which are used to purify proteins. Many of these
techniques are identical to those carried out under high performance liquid
chromatography, however use of FPLC techniques are typically for preparing
large scale batches of a purified product.
14. counter-current chromatography
Counter-current chromatography
(CCC) is a type of liquid-liquid
chromatography, where both the
stationary and mobile phases are
liquids. The operating principle of
CCC equipment requires a column
consisting of an open tube coiled
around a bobbin. The bobbin is
rotated in a double-axis gyratory
motion (a cardioid), which causes a
variable gravity (G) field to act on
(An example of HPCCC system)
the column during each rotation. This motion causes the column to see one
partitioning step per revolution and components of the sample separate in the
13. column due to their partitioning coefficient between the two immiscible liquid
phases used. There are many types of CCC available today. These include
HSCCC (High Speed CCC) and HPCCC (High Performance CCC). HPCCC is
the latest and best performing version of the instrumentation available currently.
15. chiral chromatography
Chiral chromatography involves the separation of stereo-isomers. In the case of
en-antiomers, these have no chemical or physical differences apart from being
three-dimensional mirror images. Conventional chromatography or other
separation processes are incapable of separating them. To enable chiral
separations to take place, either the mobile phase or the stationary phase must
themselves be made chiral, giving differing affinities between the
analytes. Chiral Chromatography HPLC Columns (with a chiral stationary
phase) in both normal and reversed phase are commercially available.