This document discusses applications of visible and UV spectrometry. It describes how colorimetry can be used to quantify both inorganic and organic compounds by converting colorless substances into colored derivatives using chromogenic reagents. Specific examples are provided for quantifying cations, anions, biochemical specimens, and pharmaceuticals colorimetrically. It also discusses how UV spectrometry can be used for quantitative and qualitative analysis of compounds. Methods described for quantitative analysis include using standard absorptivity values, calibration curves, and single/double point standardization. The document outlines approaches for analyzing both single component and multicomponent samples spectrophotometrically.
This document discusses supercritical fluid chromatography (SFC). SFC uses supercritical fluids like carbon dioxide as the mobile phase. Carbon dioxide is most widely used as it is non-toxic, inexpensive, and has a critical temperature and pressure that are easily reached. SFC works on the principles of adsorption and partition chromatography. It can be used to analyze and purify low to moderate weight compounds, including chiral separations. SFC instrumentation includes pumps to deliver the mobile phase, an oven for temperature control, various injectors, columns, a backpressure regulator, and detectors. SFC finds applications in fields like pharmaceuticals and has advantages over HPLC like using less toxic solvents.
High Performance Liquid Chromatography (HPLC) is described. HPLC uses high pressure to force a mobile phase through a column at a fast rate, increasing resolution. It discusses the types of chromatography used in HPLC, including normal phase, reverse phase, ion-exchange, and size-exclusion. The instrumentation of HPLC is also summarized, including components like the pump, mixing unit, degasser, injector, column, and detector.
Reversed phase chromatography is an adsorption technique used to separate nonpolar substances. It works by having a nonpolar stationary phase and a polar mobile phase, opposite of normal phase chromatography. Molecules like proteins, peptides, and nucleic acids can be separated using reversed phase chromatography. The separation depends on the hydrophobic binding of solutes from the mobile phase to the hydrophobic ligands attached to the stationary phase. Common stationary phases use silica beads with attached alkyl hydrocarbon chains of varying lengths. Gradient elution with mixtures of water and organic solvents like acetonitrile or methanol is typically used for separation. Reversed phase chromatography has applications in preparative purification of proteins, peptides, and other biomolecules.
This document discusses the instrumentation of ultraviolet-visible (UV-Vis) spectroscopy. It describes the basic components of a UV-Vis spectrophotometer including the light source, monochromator, sample cell, and detector. Common light sources for UV-Vis spectroscopy are hydrogen discharge lamps, mercury arc lamps, xenon arc lamps, and tungsten lamps. The monochromator uses either a prism or grating to filter light to the desired wavelength. Sample cells are typically made of non-reactive materials and come in matched pairs for double beam instruments. Common detectors include photomultiplier tubes, photo cells, and barrier layer cells. UV-Vis spectroscopy has applications in detecting impurities, elucidating
This document provides information about different types of columns used in high performance liquid chromatography (HPLC). It discusses normal phase and reverse phase chromatography columns. It describes various column packing materials, particle sizes, dimensions, costs and specifications. It provides details on columns from several major manufacturers like Waters, Phenomenex, Agilent, GE Healthcare and others. Preparative chromatography is also briefly mentioned. Resources for further information are listed at the end.
Industrial Applications Of Chromatography Techniquerita martin
Chromatography Technologies plays vital role in various industrial application sectors, this techniques as been used across various chemical and pharmaceutical industries. This technique is used to purify sugar from molasses, separation of enantiomers and purification of pharmaceutical proteins, pharmaceutical industry for the purification of enantiomers from racemic mixtures. Its applications were also used in food chemistry, biochemistry, petro chemistry, pharmaceutical chemistry. Also plays important roles in Purification of proteins, pharmaceuticals, fine chemicals
This document discusses applications of visible and UV spectrometry. It describes how colorimetry can be used to quantify both inorganic and organic compounds by converting colorless substances into colored derivatives using chromogenic reagents. Specific examples are provided for quantifying cations, anions, biochemical specimens, and pharmaceuticals colorimetrically. It also discusses how UV spectrometry can be used for quantitative and qualitative analysis of compounds. Methods described for quantitative analysis include using standard absorptivity values, calibration curves, and single/double point standardization. The document outlines approaches for analyzing both single component and multicomponent samples spectrophotometrically.
This document discusses supercritical fluid chromatography (SFC). SFC uses supercritical fluids like carbon dioxide as the mobile phase. Carbon dioxide is most widely used as it is non-toxic, inexpensive, and has a critical temperature and pressure that are easily reached. SFC works on the principles of adsorption and partition chromatography. It can be used to analyze and purify low to moderate weight compounds, including chiral separations. SFC instrumentation includes pumps to deliver the mobile phase, an oven for temperature control, various injectors, columns, a backpressure regulator, and detectors. SFC finds applications in fields like pharmaceuticals and has advantages over HPLC like using less toxic solvents.
High Performance Liquid Chromatography (HPLC) is described. HPLC uses high pressure to force a mobile phase through a column at a fast rate, increasing resolution. It discusses the types of chromatography used in HPLC, including normal phase, reverse phase, ion-exchange, and size-exclusion. The instrumentation of HPLC is also summarized, including components like the pump, mixing unit, degasser, injector, column, and detector.
Reversed phase chromatography is an adsorption technique used to separate nonpolar substances. It works by having a nonpolar stationary phase and a polar mobile phase, opposite of normal phase chromatography. Molecules like proteins, peptides, and nucleic acids can be separated using reversed phase chromatography. The separation depends on the hydrophobic binding of solutes from the mobile phase to the hydrophobic ligands attached to the stationary phase. Common stationary phases use silica beads with attached alkyl hydrocarbon chains of varying lengths. Gradient elution with mixtures of water and organic solvents like acetonitrile or methanol is typically used for separation. Reversed phase chromatography has applications in preparative purification of proteins, peptides, and other biomolecules.
This document discusses the instrumentation of ultraviolet-visible (UV-Vis) spectroscopy. It describes the basic components of a UV-Vis spectrophotometer including the light source, monochromator, sample cell, and detector. Common light sources for UV-Vis spectroscopy are hydrogen discharge lamps, mercury arc lamps, xenon arc lamps, and tungsten lamps. The monochromator uses either a prism or grating to filter light to the desired wavelength. Sample cells are typically made of non-reactive materials and come in matched pairs for double beam instruments. Common detectors include photomultiplier tubes, photo cells, and barrier layer cells. UV-Vis spectroscopy has applications in detecting impurities, elucidating
This document provides information about different types of columns used in high performance liquid chromatography (HPLC). It discusses normal phase and reverse phase chromatography columns. It describes various column packing materials, particle sizes, dimensions, costs and specifications. It provides details on columns from several major manufacturers like Waters, Phenomenex, Agilent, GE Healthcare and others. Preparative chromatography is also briefly mentioned. Resources for further information are listed at the end.
Industrial Applications Of Chromatography Techniquerita martin
Chromatography Technologies plays vital role in various industrial application sectors, this techniques as been used across various chemical and pharmaceutical industries. This technique is used to purify sugar from molasses, separation of enantiomers and purification of pharmaceutical proteins, pharmaceutical industry for the purification of enantiomers from racemic mixtures. Its applications were also used in food chemistry, biochemistry, petro chemistry, pharmaceutical chemistry. Also plays important roles in Purification of proteins, pharmaceuticals, fine chemicals
Instrumentation of HPLC, principle by kk sahuKAUSHAL SAHU
INTRODUCTION
Instrumentation of HPLC
TYPES OF HPLC
PARAMETERS
APPLICATION
CONCLUSION
REFERENCE
High-performance liquid chromatography ( HPLC) is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds.
HPLC mainly utilizes a column that holds packing material (stationary phase), a pump that moves the mobile phase(s) through the column, and a detector that shows the retention times of the molecules.
It includes basic knowledge about affinity chromatography along with its procedure and application. and brief description of various type of affinity chromatography.
Affinity chromatography is a method used to purify biomolecules like proteins and nucleic acids based on specific interactions between the biomolecule and a ligand immobilized on a solid support. When a mixture is passed through the column, the target biomolecule will bind to the ligand while other molecules pass through. The bound biomolecule can then be separated by changing conditions like pH or introducing a competing molecule to displace it. Affinity chromatography offers highly specific purification of target molecules in a single step.
This document presents a group presentation on chromatography given to Sir M. Aamir. The presentation introduces chromatography as a technique for separating mixtures based on the distribution of components between a mobile and stationary phase. It discusses the invention of chromatography by Michael Tswett in 1901 and important milestones. The basic components, mechanisms, classifications, and common types of chromatography are described. Uses of chromatography in forensics, medicine, research, and the pharmaceutical industry are also mentioned.
This document discusses column chromatography, which separates components of a mixture through continuous distribution between a stationary and mobile phase. It describes the basic principles, types (adsorption, partition, ion exchange, gel), components (stationary phase, mobile phase, sample), and process (elution). Column chromatography is useful for purifying compounds and isolating metabolites. It allows separation of various mixtures but requires more solvent and time than other techniques.
Column chromatography is a separation technique that uses a stationary phase, usually a solid, and a mobile liquid phase to separate mixtures. It was developed in 1900 and involves passing a liquid containing dissolved compounds through a column packed with a solid adsorbent. Components separate based on their different interactions with the stationary and mobile phases, with less strongly adsorbed compounds eluting more quickly. Column chromatography is useful for purifying compounds and isolating constituents from mixtures.
The Nobel Prize in Chemistry 1952 was awarded jointly to Archer John Porter Martin and Richard Laurence Millington Synge for their invention of partition chromatography. They developed the plate theory of chromatography, which models a chromatographic column as being divided into theoretical plates with each plate representing equilibrium between the mobile and stationary phases. The number of theoretical plates is used to represent the efficiency and performance of the column.
Chiral chromatography & ion pair chromatographyHemantBansode2
This document discusses chiral chromatography and ion-pair chromatography. It begins with definitions of stereoisomers including enantiomers, diastereomers, and racemates. It then covers the principles of chiral chromatography, which involves using a chiral stationary phase or chiral derivatization to separate enantiomers. Applications include resolving drug enantiomers. The document also discusses ion-pair chromatography, which uses ion-pairing reagents to separate charged analytes in HPLC. Key factors that affect ion-pair chromatography are also reviewed.
Introduction to chromatography, Definition of Chromatography, Types of column chromatography, Theory of chromatography, Practical considerations in column chromatography , Factors affecting efficiency of a column, Applications.
Detectors are devices used in gas chromatography and liquid chromatography to detect components of mixtures being analyzed. There are two main types of detectors: destructive and non-destructive. Destructive detectors transform the analyte through burning, evaporation, or mixing with reagents before measurement, such as the flame ionization detector (FID) and nitrogen phosphorus detector (NPD). Non-destructive detectors directly measure properties like UV absorption or thermal conductivity without transforming the analyte, exemplified by the thermal conductivity detector. The most commonly used detectors are the FID, NPD, and TCD due to their high sensitivity, reproducibility, and selectivity for certain compounds.
This document provides an overview of ion exchange chromatography. It defines ion exchange chromatography as a process that separates similar charged ions using an ion exchange resin. The document then classifies resins, describes the principles and apparatus of ion exchange chromatography, and lists some key factors that affect resolution. It also outlines several applications of ion exchange chromatography such as separating metal ions, analyzing water samples, and purifying biochemical compounds.
Gel permeation chromatography (GPC) separates molecules based on size as they pass through a column containing porous gel beads. Larger molecules pass through the column faster than smaller ones that penetrate the gel pores. Key components of GPC include the stationary phase gel beads, mobile phase solvent, columns, pump, and detectors. GPC is useful for fractionating, purifying, and determining the molecular weights of polymers, proteins, and other biomolecules.
Column chromatography is a separation technique that uses a column packed with a stationary phase and a liquid mobile phase. Components of a mixture are separated as they travel through the column at different rates depending on how strongly they interact with the stationary phase. Factors that affect column chromatography include the type of stationary phase, particle size, flow rate of the mobile phase, column temperature, and concentration of the mixture. Column chromatography is commonly used to purify compounds in chemistry and biochemistry.
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.
1) IR spectroscopy uses infrared radiation to identify chemical substances by their absorption patterns.
2) The main components of an IR spectrometer are a radiation source, monochromator, sample cells, detectors, and recorder.
3) Common radiation sources are Nernst glowers, globar sources, and incandescent wires, which emit IR radiation that is focused through the sample.
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.
Theory of high performance liquid chromatography pptshweta more
This document provides an overview of the theory of high performance liquid chromatography (HPLC). It discusses key concepts such as the retention factor (k), which is a measure of how long a compound is retained on the column. Selectivity (α) refers to the ability to distinguish between sample components, and is calculated as a ratio of the k values. Resolution (Rs) is the most important measure of separation, and depends on factors like k, α, and the number of theoretical plates (N). N is a measure of column efficiency, and the height equivalent of a theoretical plate (HETP) describes efficiency. The document outlines how these parameters can be optimized to improve separation and resolution.
HPLC is a form of liquid chromatography used to separate compounds dissolved in solution based on how they partition between a stationary and mobile phase. The key components of an HPLC system are a pump, injector, column, and detector. Method development involves selecting parameters like the mobile phase, column type, detection method, and chromatography conditions to optimize separation of the sample components. HPLC offers advantages like high sensitivity, rapid analysis times, and use for both analytical qualitative and quantitative analysis.
HPLC is a type of liquid chromatography that can separate mixtures of chemicals. It works by pumping a pressurized liquid solvent (mobile phase) through a column containing a solid material (stationary phase). Samples are injected and the different compounds interact differently with the phases, causing them to elute from the column at different retention times, allowing separation. HPLC has advantages over other methods like higher separation efficiency, reproducibility, and ability to analyze a wide range of compounds dissolved in liquid. It is used in various fields like medicine, food, environment, and industry.
Instrumentation of HPLC, principle by kk sahuKAUSHAL SAHU
INTRODUCTION
Instrumentation of HPLC
TYPES OF HPLC
PARAMETERS
APPLICATION
CONCLUSION
REFERENCE
High-performance liquid chromatography ( HPLC) is a specific form of column chromatography generally used in biochemistry and analysis to separate, identify, and quantify the active compounds.
HPLC mainly utilizes a column that holds packing material (stationary phase), a pump that moves the mobile phase(s) through the column, and a detector that shows the retention times of the molecules.
It includes basic knowledge about affinity chromatography along with its procedure and application. and brief description of various type of affinity chromatography.
Affinity chromatography is a method used to purify biomolecules like proteins and nucleic acids based on specific interactions between the biomolecule and a ligand immobilized on a solid support. When a mixture is passed through the column, the target biomolecule will bind to the ligand while other molecules pass through. The bound biomolecule can then be separated by changing conditions like pH or introducing a competing molecule to displace it. Affinity chromatography offers highly specific purification of target molecules in a single step.
This document presents a group presentation on chromatography given to Sir M. Aamir. The presentation introduces chromatography as a technique for separating mixtures based on the distribution of components between a mobile and stationary phase. It discusses the invention of chromatography by Michael Tswett in 1901 and important milestones. The basic components, mechanisms, classifications, and common types of chromatography are described. Uses of chromatography in forensics, medicine, research, and the pharmaceutical industry are also mentioned.
This document discusses column chromatography, which separates components of a mixture through continuous distribution between a stationary and mobile phase. It describes the basic principles, types (adsorption, partition, ion exchange, gel), components (stationary phase, mobile phase, sample), and process (elution). Column chromatography is useful for purifying compounds and isolating metabolites. It allows separation of various mixtures but requires more solvent and time than other techniques.
Column chromatography is a separation technique that uses a stationary phase, usually a solid, and a mobile liquid phase to separate mixtures. It was developed in 1900 and involves passing a liquid containing dissolved compounds through a column packed with a solid adsorbent. Components separate based on their different interactions with the stationary and mobile phases, with less strongly adsorbed compounds eluting more quickly. Column chromatography is useful for purifying compounds and isolating constituents from mixtures.
The Nobel Prize in Chemistry 1952 was awarded jointly to Archer John Porter Martin and Richard Laurence Millington Synge for their invention of partition chromatography. They developed the plate theory of chromatography, which models a chromatographic column as being divided into theoretical plates with each plate representing equilibrium between the mobile and stationary phases. The number of theoretical plates is used to represent the efficiency and performance of the column.
Chiral chromatography & ion pair chromatographyHemantBansode2
This document discusses chiral chromatography and ion-pair chromatography. It begins with definitions of stereoisomers including enantiomers, diastereomers, and racemates. It then covers the principles of chiral chromatography, which involves using a chiral stationary phase or chiral derivatization to separate enantiomers. Applications include resolving drug enantiomers. The document also discusses ion-pair chromatography, which uses ion-pairing reagents to separate charged analytes in HPLC. Key factors that affect ion-pair chromatography are also reviewed.
Introduction to chromatography, Definition of Chromatography, Types of column chromatography, Theory of chromatography, Practical considerations in column chromatography , Factors affecting efficiency of a column, Applications.
Detectors are devices used in gas chromatography and liquid chromatography to detect components of mixtures being analyzed. There are two main types of detectors: destructive and non-destructive. Destructive detectors transform the analyte through burning, evaporation, or mixing with reagents before measurement, such as the flame ionization detector (FID) and nitrogen phosphorus detector (NPD). Non-destructive detectors directly measure properties like UV absorption or thermal conductivity without transforming the analyte, exemplified by the thermal conductivity detector. The most commonly used detectors are the FID, NPD, and TCD due to their high sensitivity, reproducibility, and selectivity for certain compounds.
This document provides an overview of ion exchange chromatography. It defines ion exchange chromatography as a process that separates similar charged ions using an ion exchange resin. The document then classifies resins, describes the principles and apparatus of ion exchange chromatography, and lists some key factors that affect resolution. It also outlines several applications of ion exchange chromatography such as separating metal ions, analyzing water samples, and purifying biochemical compounds.
Gel permeation chromatography (GPC) separates molecules based on size as they pass through a column containing porous gel beads. Larger molecules pass through the column faster than smaller ones that penetrate the gel pores. Key components of GPC include the stationary phase gel beads, mobile phase solvent, columns, pump, and detectors. GPC is useful for fractionating, purifying, and determining the molecular weights of polymers, proteins, and other biomolecules.
Column chromatography is a separation technique that uses a column packed with a stationary phase and a liquid mobile phase. Components of a mixture are separated as they travel through the column at different rates depending on how strongly they interact with the stationary phase. Factors that affect column chromatography include the type of stationary phase, particle size, flow rate of the mobile phase, column temperature, and concentration of the mixture. Column chromatography is commonly used to purify compounds in chemistry and biochemistry.
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.
1) IR spectroscopy uses infrared radiation to identify chemical substances by their absorption patterns.
2) The main components of an IR spectrometer are a radiation source, monochromator, sample cells, detectors, and recorder.
3) Common radiation sources are Nernst glowers, globar sources, and incandescent wires, which emit IR radiation that is focused through the sample.
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.
Theory of high performance liquid chromatography pptshweta more
This document provides an overview of the theory of high performance liquid chromatography (HPLC). It discusses key concepts such as the retention factor (k), which is a measure of how long a compound is retained on the column. Selectivity (α) refers to the ability to distinguish between sample components, and is calculated as a ratio of the k values. Resolution (Rs) is the most important measure of separation, and depends on factors like k, α, and the number of theoretical plates (N). N is a measure of column efficiency, and the height equivalent of a theoretical plate (HETP) describes efficiency. The document outlines how these parameters can be optimized to improve separation and resolution.
HPLC is a form of liquid chromatography used to separate compounds dissolved in solution based on how they partition between a stationary and mobile phase. The key components of an HPLC system are a pump, injector, column, and detector. Method development involves selecting parameters like the mobile phase, column type, detection method, and chromatography conditions to optimize separation of the sample components. HPLC offers advantages like high sensitivity, rapid analysis times, and use for both analytical qualitative and quantitative analysis.
HPLC is a type of liquid chromatography that can separate mixtures of chemicals. It works by pumping a pressurized liquid solvent (mobile phase) through a column containing a solid material (stationary phase). Samples are injected and the different compounds interact differently with the phases, causing them to elute from the column at different retention times, allowing separation. HPLC has advantages over other methods like higher separation efficiency, reproducibility, and ability to analyze a wide range of compounds dissolved in liquid. It is used in various fields like medicine, food, environment, and industry.
Chromatography is a method used to separate mixtures by distributing components between a stationary and mobile phase. There are several types including thin layer chromatography (TLC), which separates compounds on plates coated with adsorbent. Column chromatography packs an absorbent in a column and elutes components with a solvent. High performance liquid chromatography (HPLC) uses high pressure to force components through a column at high speed. Chromatography techniques are used in forensic analysis and research to identify substances.
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 technique used to separate mixtures based on how their components interact with both a mobile and stationary phase. It was first developed in 1900 by Russian scientist Mikhail Tsvet to separate plant pigments. There are several types of chromatography that differ based on the phases used, including paper chromatography, thin layer chromatography, gas chromatography, ion exchange chromatography, gel filtration chromatography, and affinity chromatography. High performance liquid chromatography is a modern technique that uses small particle sizes and high pressure to improve separation efficiency.
A review on ipce and pec measurements and materials p.basnetPradip Basnet
The slides show how to measure the photoelectrochemical (PEC) properties of a light-active photocatalyst (usually semiconductor) and current literature summary for water splitting using sunlight.
HPLC Principle,Instrumentation and ApplicationAlakesh Pradhan
HPLC Chromatography and its principle
Liquid chromatography
High Performance Liquid Chromatography ( HPLC )
The components of the high performance liquid chromatograph (HPLC).
The separation process.
The chromatogram
Waves are disturbances that transfer energy through a medium from one point to another. They can be transverse waves, where the disturbance is perpendicular to the direction of travel, or longitudinal waves, where the disturbance is parallel. Waves have properties like wavelength, amplitude, frequency, and speed. The speed of a wave depends on its frequency and wavelength and can be calculated using the formula that the speed equals the frequency multiplied by the wavelength. Waves undergo behaviors like reflection, refraction, and diffraction as they interact with barriers and changes in their medium.
Chromatography is a technique used to separate mixtures by distributing components between two phases - a stationary phase and a mobile phase. The mixture is dissolved in the mobile phase which carries it through a column containing the stationary phase. Components travel at different speeds depending on how they partition between the phases, allowing separation. Chromatography can be used for analytical purposes to determine the presence and proportions of components, or preparatively to purify components for further use. Key terms include the mobile phase, stationary phase, retention time, and resolution.
Chromatography is a technique used to separate mixtures into individual components. There are several types of chromatography including paper chromatography, column chromatography, and thin layer chromatography. Paper chromatography uses a stationary phase like cellulose and a mobile phase like solvents to separate components by capillary action. Column chromatography uses a column packed with an adsorbent as the stationary phase and a liquid mobile phase to separate components based on differences in affinity. Thin layer chromatography coats an adsorbent onto a glass plate as the stationary phase and uses a liquid mobile phase to separate components traveling different distances based on affinity.
This document discusses the use of chromatography techniques to separate plant constituents. It begins with background on pharmacognosy and why extraction of therapeutic vs non-therapeutic constituents is important. Chromatography techniques like thin layer chromatography, gas chromatography, and HPLC are described for separating mixtures based on differences in compounds' affinities for mobile vs stationary phases. Specific discussion is provided on separating alkaloids, volatile oils, and sugars using appropriate solvent systems and chromatographic conditions.
RADAR - RAdio Detection And Ranging
This is the Part 2 of 2 of RADAR Introduction.
For comments please contact me at solo.hermelin@gmail.com.
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This document provides an overview of high performance liquid chromatography (HPLC). It discusses the key components of an HPLC system including the solvent rack, pump, injector, separation column, and detector. It also describes different chromatography techniques such as normal phase chromatography, reversed phase chromatography, and ion exchange chromatography which separate compounds based on polarity, electrical charge, and molecular size. Finally, it discusses factors that influence separation such as stationary phase particle size and eluent composition.
Gas chromatography is a technique used to separate components of a vaporized sample. It works by partitioning the components between a mobile gaseous phase and a stationary phase within a column. The sample is injected and vaporized, then transported through the column by the mobile phase gas. As the components pass through the column they are separated and detected. Common detectors include the flame ionization detector (FID), thermal conductivity detector (TCD), and electron capture detector (ECD). The FID responds to organic compounds, the TCD is universal but less sensitive, and the ECD selectively detects halogen-containing compounds.
This document provides an overview of gas chromatography (GC). It describes the basic components and principles of how GC works, including the carrier gas, injection port, separation column in an oven, and detector. It explains that GC separates volatile organic compounds based on how they partition between the mobile gas phase and stationary liquid phase. The document also outlines different types of columns, sample preparation procedures, common detectors like the flame ionization detector, and applications of GC in fields like environmental analysis and refineries.
Here are the key principles of the different chromatography techniques covered in the document:
- Paper chromatography relies on the differential migration rates of compounds through a stationary phase (filter paper) based on their varying interactions with the mobile phase solvent. Components separate based on differences in solubility and affinity for the mobile vs. stationary phases.
- Thin layer chromatography uses a thin stationary phase coating (e.g. silica gel) on a plate. Components separate based on differences in their partitioning behavior between the mobile liquid phase and stationary solid phase during capillary movement.
- Column chromatography uses a packed stationary phase within a column. Components separate based on differences in their distribution between the stationary phase adsorbent and percolating mobile phase
BASICS OF LASER AND IT'S USE IN DERMATOLOGYRohit Singh
The document discusses lasers and their uses in dermatology. It begins with definitions and a brief history of lasers, describing some important early pioneers and dates. The basic components and working principles of lasers are then explained, including population inversion, stimulated emission, and the use of gain medium, pumping systems, and optical resonators. Different types of lasers are also categorized based on their gain medium, such as gas, solid state, and dye lasers. Applications of lasers in dermatology are enabled by their interactions with chromophores in the skin and ability to penetrate at varying depths depending on the wavelength. Thermal effects on tissue include photocoagulation and photo-vaporization.
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.
APPLICATIONS OF GAS CHROMATOGRAPHY [APPLICATIONS OF GC] BY Prof. Dr. P.RAVISA...Dr. Ravi Sankar
Gas chromatography (GC) is an instrumental technique used for drug analysis, toxicology, and identification of organic compounds. It allows for high-speed and sensitive analysis of mixtures like pesticides. GC coupled with mass spectrometry is used in applications like pharmaceutical analysis, food safety testing, and indoor air quality monitoring. It is also used in forensic sciences to identify drugs and other compounds.
The document discusses hyphenated analytical techniques, specifically gas chromatography-mass spectrometry (GC-MS). It outlines the advantages of hyphenated techniques, describes the instrumentation and interface of GC-MS, and lists several applications including metabolite profiling, detection of compounds in plant tissues, analysis of aromatic amines, identification of volatile components, and environmental and forensic analysis. GC-MS is presented as a powerful technique for the sensitive analysis of various compounds.
Gas chromatography is a technique used to separate and analyze compounds that can be vaporized without decomposing. It works by carrying a gaseous or vaporized sample mixture through a column via an inert gas mobile phase. Components interact differently with the stationary phase coating the column and exit at different retention times, allowing separation. Common applications include analyzing purity, identifying unknown compounds, and preparing pure samples. Advantages include high sensitivity and resolution, while disadvantages include limited sample types and inability to recover individual components.
ANALYSIS THROUGH chromatography techniques.pptxRashmiSanghi1
Chromatography is a technique used to separate chemical components in a complex mixture. It works by carrying components through a stationary phase at different rates using a mobile phase, usually liquid or gas. Liquid chromatography uses high pressure to push a liquid mobile phase through a column, while gas chromatography uses an inert gas and higher temperatures. Different detectors can be used to analyze the separated components as they exit the column. Common detectors measure properties like thermal conductivity, ionization, or light emission to identify the separated chemicals. Chromatography is a powerful analytical and preparative separation method.
HPLC uses a liquid mobile phase to transport analytes through a column packed with a stationary phase. It can analyze non-volatile and thermally fragile compounds over a wide range of molecular weights. HPLC is preferable to GC when samples cannot be analyzed without lengthy preparation due to issues with volatility. The key components of an HPLC system are the pump, injector, column, and detector. Various parameters like mobile phase composition, flow rate, and temperature must be optimized to generate a satisfactory chromatogram.
The document discusses gas chromatography. It begins by providing a brief history of chromatography and describing the basic components and process of gas chromatography. It then discusses the types of gas chromatography, including gas-solid and gas-liquid, and describes the typical instrumentation used, including carrier gases, sample injection systems, columns, and common detectors like FID, TCD, ECD, and TID. Applications of gas chromatography include qualitative and quantitative analysis of organic compounds. Advantages are its high resolution and sensitivity, while disadvantages include its limitation to volatile samples.
Gas chromatography is an analytical technique used to separate mixtures by vaporizing the components and passing them through a column with a mobile gas phase and a stationary liquid phase. It was pioneered in the 1940s and the first gas chromatograph was developed in 1951. A typical gas chromatography system consists of a gas inlet, injector, column inside an oven, detector, and data system. The sample is injected and separated in the column based on interactions between the phases, then detected and analyzed to produce a chromatogram showing the composition of the original mixture.
This document provides an introduction and overview of gas chromatography (GC). It discusses the basic principles of GC, which involves separating components of a mixture based on how they partition between a stationary and mobile phase. The key components of a GC system are described, including the injector where samples are introduced, the column where separation occurs, the oven that controls temperature, and various detectors. Different types of columns, stationary phases, temperature programs, and detectors are discussed to provide flexibility in GC analysis for a wide range of applications.
Gas chromatography is a technique used to separate mixtures by distributing components between a stationary and mobile phase. It can separate volatile organic compounds without decomposition. There are two main types: gas-solid and gas-liquid chromatography. The instrument uses a carrier gas to move samples through a column where separation occurs based on partitioning between the column coating and gas. Detectors then produce signals for separated components. Common detectors include FID, TCD, ECD, and FPD. Gas chromatography finds applications in fields like food analysis and environmental monitoring due to its speed, sensitivity, and ability to separate complex mixtures both qualitatively and quantitatively.
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 overview of gas chromatography. It begins by defining chromatography and tracing the history of gas chromatography from its origins in 1903 to its development in the 1940s-1950s. It then describes the basic components and working principles of gas chromatography, including the mobile phase, stationary phase, factors that influence separation, and common instrumentation. It also discusses different types of chromatography techniques and gas chromatography columns. In summary, the document provides a comprehensive introduction to gas chromatography, its history, principles, instrumentation and applications.
Chromatography is a technique used to separate mixtures by distributing components between two phases, stationary and mobile. The mixture is dissolved in a mobile phase that carries it through a column containing a stationary phase. Components travel at different rates based on how they partition between the phases, allowing separation. Common chromatography methods include gas chromatography, liquid chromatography, and thin layer chromatography. Chromatography has applications in identifying unknown substances like drugs, proteins, and plant pigments. It was first developed in 1903 and continues to be an important analytical technique.
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.
Gas chromatography (GC) is an analytical technique used to separate and detect the chemical components of a sample mixture to determine their presence or absence and/or quantities.Gas chromatography is usually used to separate and measure organic molecules and gases. For the technique to function, the components being analyzed must be volatile, be thermally stable, and have a molecular weight of below 1250 Da.Gas Chromatography involves the use of a separation column, which is made from a length of glass, fused silica, or metal tubing.Gas chromatography is a novel technique for separating and quantitating vaporized compounds using an inert carrier gas. It operates on similar principles to column permeation chromatography, where a sample is dissolved in a mobile phase and passed through a porous stationary structure
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.
This document discusses gas chromatography (GC), a popular chromatography technique used to separate volatile compounds based on how they partition between a gaseous mobile phase and a stationary solid or liquid phase. GC was invented in 1901 by Russian botanist Mikhail Tswett to separate plant pigments. It works by vaporizing a sample and carrying it through a column with an inert carrier gas, where different compounds interact differently with the stationary phase and elute out at different retention times due to differences in their properties. The document outlines the basic principles and components of a GC system.
Gas chromatography (GC) is a common type of chromatography used in analytical chemistry for separating and analysing compounds that can be vaporised without decomposition.
GASSCHROMATOGRAPHY, ADVANCED STUDY OF THE FOLLOWING AND THEIR APPLICATIONS, I...Dr. Ravi Sankar
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BY P.RAVISANKAR, VIGNAN PHARMACY COLLEGE, VADLAMUDI, GUNTUR, ANDHRA PRADESH, INDIA.
The document discusses different types of chromatography. It begins with an introduction to chromatography, including its history and principles. It then describes various classifications of chromatography based on mechanism and phases. Specific techniques are defined, including adsorption chromatography, partition chromatography, gas-liquid chromatography, solid-liquid chromatography, and liquid-liquid chromatography. Key terms are explained. Applications and steps of chromatographic separation are outlined. Important properties of liquid stationary phases are also summarized.
This document provides an overview of chromatographic techniques. It begins with definitions and a brief history, then covers principles, applications, classification, specific techniques (e.g. gas chromatography, liquid chromatography), terms, and properties of stationary phases. The document presents chromatographic methods and their use in separating mixtures like drugs, proteins, and other compounds. It concludes that supercritical fluid chromatography falls between HPLC and GC in performance for applications in pharmaceutical and bioanalytical analysis.
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Chapter wise All Notes of First year Basic Civil Engineering.pptxDenish Jangid
Chapter wise All Notes of First year Basic Civil Engineering
Syllabus
Chapter-1
Introduction to objective, scope and outcome the subject
Chapter 2
Introduction: Scope and Specialization of Civil Engineering, Role of civil Engineer in Society, Impact of infrastructural development on economy of country.
Chapter 3
Surveying: Object Principles & Types of Surveying; Site Plans, Plans & Maps; Scales & Unit of different Measurements.
Linear Measurements: Instruments used. Linear Measurement by Tape, Ranging out Survey Lines and overcoming Obstructions; Measurements on sloping ground; Tape corrections, conventional symbols. Angular Measurements: Instruments used; Introduction to Compass Surveying, Bearings and Longitude & Latitude of a Line, Introduction to total station.
Levelling: Instrument used Object of levelling, Methods of levelling in brief, and Contour maps.
Chapter 4
Buildings: Selection of site for Buildings, Layout of Building Plan, Types of buildings, Plinth area, carpet area, floor space index, Introduction to building byelaws, concept of sun light & ventilation. Components of Buildings & their functions, Basic concept of R.C.C., Introduction to types of foundation
Chapter 5
Transportation: Introduction to Transportation Engineering; Traffic and Road Safety: Types and Characteristics of Various Modes of Transportation; Various Road Traffic Signs, Causes of Accidents and Road Safety Measures.
Chapter 6
Environmental Engineering: Environmental Pollution, Environmental Acts and Regulations, Functional Concepts of Ecology, Basics of Species, Biodiversity, Ecosystem, Hydrological Cycle; Chemical Cycles: Carbon, Nitrogen & Phosphorus; Energy Flow in Ecosystems.
Water Pollution: Water Quality standards, Introduction to Treatment & Disposal of Waste Water. Reuse and Saving of Water, Rain Water Harvesting. Solid Waste Management: Classification of Solid Waste, Collection, Transportation and Disposal of Solid. Recycling of Solid Waste: Energy Recovery, Sanitary Landfill, On-Site Sanitation. Air & Noise Pollution: Primary and Secondary air pollutants, Harmful effects of Air Pollution, Control of Air Pollution. . Noise Pollution Harmful Effects of noise pollution, control of noise pollution, Global warming & Climate Change, Ozone depletion, Greenhouse effect
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1. Palancharmy, Basic Civil Engineering, McGraw Hill publishers.
2. Satheesh Gopi, Basic Civil Engineering, Pearson Publishers.
3. Ketki Rangwala Dalal, Essentials of Civil Engineering, Charotar Publishing House.
4. BCP, Surveying volume 1
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3. Contents Page No.
Water Analysis……………………………………......................04
Organic Trace Pollutants………………………………………...05
History of Chromatography……………………………………..07
Definition of Chromatography………………………………....10
Principles of Chromatography…………………………………...11
Paper Chromatographic Technique. ……………………………..12
Gas Chromatography (GC)………………………………………14
Principle of Gas Chromatography (GC)…………………………15
High Performance Liquid Chromatography (HPLC)……………19
The differences between HPLC & GC…………………………..23
The separation process…………………………………………..29
The Chromatogram……………………………………………..46
HPLC Applications………………………………………….…..49
References……………………………………………………….50
3
4. A chemical analysis of a water solution in which specific ions
and their concentrations are determined and recorded.
The character of the water solution then can be described in
terms of the individual ion concentrations and the total dissolved
solids, in units of ppm or mg/liter.
A complete analysis will include measurement
of pH, hardness, and bacteriological testing.
For limits of these criteria recommended for good quality
domestic water, suggested by U.S. Environmental Protection
Agency (EPA), consult EPA 822-R-94-001, May 1994 or CSU.
4
5. Include
Naturally occurring compounds from decomposition of
OM
Anthropogenic pollutants
Degradation and inter-reaction products of pollutants
Substances derived from sewage treatment
5
Typical analysis:
Individual compounds or groups of compounds
Total analysis of all organic components
Field screening for specific pollutants prior to lab analysis
Qualitative identification of trade products in spills and
discharges
Organic Trace Pollutants (OTP)
7. Chromatography
The word “CHORMATOGRAPHY” was suggested by a Russain
Scientist, Michael Tswett in 1906.
M. Tswett was the first to use the term "chromatography" derived
from two Greek words "Chroma" meaning color and "graphein"
meaning to write.
The technique of paper chromatography was introduced into
biological research by Martin and Synge in 1941.
7
8. 8
1901 - invented chromatography
1903 - Mikhail Tswett separated plant pigments
using paper chromatography
liquid-solid chromatography
1930’s - Schuftan & Eucken use vapor as the
mobile phase
gas solid chromatography
1941 - paper chromatography was introduced into
biological research by Martin and Synge.
History of Chromatography
9. Invention of Chromatography
Mikhail Tswett invented
chromatography in 1901
during his research on
plant pigments.
He used the technique to
separate various plant
pigments such as
chlorophylls, xanthophylls
and carotenoids. Mikhail Tswett
Russian Botanist
(1872-1919) 9
10. Definition of chromatography
Tswett (1906) stated that " chromatography is a method
in which the components of a mixture are separated on
adsorbent column in a flowing system”.
IUPAC definition (International Union of pure and
applied Chemistry) (1993):
Chromatography is a physical method of separation in
which the components to be separated are distributed
between two phases, one of which is stationary while the
other moves in a definite direction.
The stationary phase may be a solid, or a liquid supported
on a solid or gel, the mobile phase may be either a gas or a
liquid.
10
11. Principles of Chromatography
Chromatography is a physical process.
Any Chromatography system is composed of three
Components :
Stationary phase
Mobile phase
Mixture to be separated
We can only control stationary and mobile phase as
mixtures are the problem we have to deal with.
Chromatography is a dynamic process in which the
mobile phase moves in definite direction. 11
12. Flow sheet for the use of Paper Chromatographic Technique
Pour the solvent system
into the petriplates
Apply the sample in the
centre of the filter paper
Place the filter paper
between the plates
Run the Chromatogram
till the end of paper
Visualization of spots
Calculate the Rf Value
Air Dry
Rf Value:
Define as the ratio of the
distance traveled by a given
compound as compound to
the distance traveled by the
solvent.
12
14. Gas Chromatography (GC)
Gas chromatography is a chromatographic technique
that can be used to separate volatile organic compounds.
It consists of
a flowing mobile phase
an injection port
a separation column (the stationary phase)
an oven
a detector.
14
15. Principle Gas Chromatography
The organic compounds are separated due to
differences in their partitioning behavior between the
mobile gas phase and the stationary phase in the
column.
Mobile phases are generally inert gases such as helium,
argon, or nitrogen.
The injection port consists of a rubber septum through
which a syringe needle is inserted to inject the sample.
The injection port is maintained at a higher temperature than
the boiling point of the least volatile component in the sample
mixture.
Cont…
15
16. Since the partitioning behavior is dependent on
temperature, the separation column is usually
contained in a thermostat-controlled oven.
Separating components with a wide range of boiling
points is accomplished by starting at a low oven
temperature and increasing the temperature over time
to elute the high-boiling point components.
Principle GC
16
20. The differences between High Performance Liquid
Chromatography and Gas Chromatography.
The components of the high performance liquid
chromatograph (HPLC).
The separation process.
The chromatogram.
The most common modes of HPLC.
20
In This Section, We Will Discuss
21. 21
I need a quantitative
separation of
carbohydrates in some
of our products
as soon as possible.
I’ll need a separation
technique.
I’ll get
on it!
You’ve Got a Problem to Solve
22. 22
I have two separation techniques in my lab,
High Performance Liquid Chromatography
and Gas Chromatography. Which should I use?
Separation Techniques
23. 23
Sample Volatility Sample Polarity
HPLC
•No volatility requirement
•Sample must be soluble
in mobile phase
GC
•Sample must be volatile
HPLC
GC
•Separates both polar and
non polar compounds
•PAH - inorganic ions
•Samples are nonpolar
and polar
Comparison of HPLC and GC
25. 25
Sample Thermal Lability Sample Molecular Weight
HPLC
•Analysis can take place
at or below room
temperature
GC
•Sample must be able
to survive high
temperature injection
port and column
HPLC
GC
•No theoretical upper limit
•In practicality, solubility is
limit.
•Typically < 500 amu
Comparison of HPLC and GC
26. 26
Sample Preparation Sample Size
HPLC
•Sample must be filtered
•Sample should be in
same solvent as mobile
phase
GC
•Solvent must be volatile
and generally lower
boiling than analytes
HPLC
GC
•Sample size based upon
column i.d.
•Typically 1 - 5 L
Comparison of HPLC and GC
27. 27
Separation Mechanism Detectors
HPLC
•Both stationary phase
and mobile phase take
part
GC
•Mobile phase is a
sample carrier only
HPLC
GC
•Most common UV-Vis
•Wide range of non-
destructive detectors
•3-dimensional detectors
•Sensitivity to fg (detector
dependent)
•Most common FID,
universal to organic
compounds
Comparison of HPLC and GC
28. 28
Carbohydrates
1. fructose
2. Glucose
3. Saccharose
4. Palatinose
5. Trehalulose
6. isomaltose
Zorbax NH2 (4.6 x 250 mm)
70/30 Acetonitrile/Water
1 mL/min Detect=Refractive Index
1
2
3
4
5
mAU
time
6
How can We Analyze the Sample?
29. Separations
29
Separation in based upon differential
migration between the stationary and
mobile phases.
Stationary Phase - the phase which
remains fixed in the column, e.g. C18,
Silica
Mobile Phase - carries the sample
through the stationary phase as it
moves through the column.
Injector
Detector
Column
Solvents
Mixer
Pumps
High Performance Liquid Chromatograph
Waste