Chromatography is a technique used to separate mixtures by exploiting differences in how components interact with stationary and mobile phases. It was discovered in 1906 by Tswett and is commonly used for separation, purification, identification, and quantification of mixture components. The key components are a stationary phase, mobile phase, and sample mixture. Different types of chromatography exist depending on the phase types, including solid-liquid, liquid-liquid, gas-liquid, and liquid-solid. Common applications include analyzing pharmaceuticals, detecting substances in blood/tissue, forensic analysis, and environmental monitoring.
Sandesh Walke completed a chemistry investigatory project on paper chromatography. He thanks his teacher Ms. Dhruva Pardeshi for guiding the project and his principal for providing facilities. Paper chromatography is a technique that separates mixtures by their different migration rates on chromatographic paper. Key aspects covered include sample application, development with a mobile solvent phase, and calculation of Rf values to quantify separation. Paper chromatography has various applications like separation of mixtures, purity testing, and analysis of foods, drugs and other samples.
Paper chromatography is a technique used to separate mixtures into individual components. It works by allowing the molecules in a mixture to distribute themselves between a stationary phase (filter paper) and a mobile phase (solvent). Different molecules will travel different distances up the paper depending on how strongly they interact with each phase. Paper chromatography has been replaced in many applications by thin layer chromatography, which provides faster runs and better separation. However, paper chromatography is still useful for teaching chromatography principles in educational settings.
Chromatography is a technique used to separate mixtures into individual components. It works by using the differential affinity and solubility of components for both a mobile phase that carries the mixture and a stationary phase that the mixture passes through. The separation of components depends on how strongly they interact with each phase. Common types of chromatography include liquid chromatography, gas chromatography, paper chromatography, and thin-layer chromatography. Paper chromatography can be used to separate the dyes in markers by running samples along a strip with an increasing solvent mixture as the mobile phase.
Paper chromatography is a technique used to separate colored chemicals or substances. It involves a stationary phase, usually a polar substance like water held in paper, and a mobile phase, generally a non-polar solvent mixture, that travels up the stationary phase through capillary action. Sample mixtures are spotted on the paper and separated as their components partition between the two phases based on differences in their affinity. Variations include ascending or descending based on solvent direction, and two-dimensional using two solvents rotated 90 degrees. The separation is analyzed by calculating Rf values, the ratio of compound travel distance to solvent front.
The versatile instrument is used to isolate unknown compounds from a HPTLC/TLC plate and transfer them into a mass spectrometer for identification or structure elucidation.
This document provides an overview of paper chromatography. It discusses the history and types of paper chromatography, including the principles of separation. It describes the practical requirements like the stationary and mobile phases used. It explains how to calculate Rf and Rx values and factors that can affect them. Various detection methods are outlined as well as sources of error. Finally, the applications and advantages of paper chromatography are highlighted.
This document provides information on paper chromatography, including its history, key terms, processes, techniques, factors that affect separation, and applications. Paper chromatography is an analytical technique used to separate mixtures based on differences in how compounds partition between a stationary phase, such as the paper, and a mobile phase, such as a solvent. Key points covered include the different types of paper chromatography, factors that influence separation like the solvent used, and how paper chromatography can be used to analyze organic compounds.
HPLC[ HIGH PERPROMANCE LIQUID CHROMATOGRAPHY OR HIGH PRESSURE LIQUID CHROMAT...Dr. Ravi Sankar
GASSCHROMATOGRAPHY[GC], ADVANCED STUDY OF THE FOLLOWING AND THEIR APPLICATIONS, INTRODUCTION, THEORY, COLUMN OPERATION,INSTRUMENTATION AND DETECTION,APPLICATIONS AND ADVANTAGES OF GC,PRINCIPLE OF SEPARATION IN GC, HOW GC MECHINE WORKS? COLUMN, DETECTORS.
BY P.RAVISANKAR, VIGNAN PHARMACY COLLEGE, VADLAMUDI, GUNTUR, A.P, INDIA.
Sandesh Walke completed a chemistry investigatory project on paper chromatography. He thanks his teacher Ms. Dhruva Pardeshi for guiding the project and his principal for providing facilities. Paper chromatography is a technique that separates mixtures by their different migration rates on chromatographic paper. Key aspects covered include sample application, development with a mobile solvent phase, and calculation of Rf values to quantify separation. Paper chromatography has various applications like separation of mixtures, purity testing, and analysis of foods, drugs and other samples.
Paper chromatography is a technique used to separate mixtures into individual components. It works by allowing the molecules in a mixture to distribute themselves between a stationary phase (filter paper) and a mobile phase (solvent). Different molecules will travel different distances up the paper depending on how strongly they interact with each phase. Paper chromatography has been replaced in many applications by thin layer chromatography, which provides faster runs and better separation. However, paper chromatography is still useful for teaching chromatography principles in educational settings.
Chromatography is a technique used to separate mixtures into individual components. It works by using the differential affinity and solubility of components for both a mobile phase that carries the mixture and a stationary phase that the mixture passes through. The separation of components depends on how strongly they interact with each phase. Common types of chromatography include liquid chromatography, gas chromatography, paper chromatography, and thin-layer chromatography. Paper chromatography can be used to separate the dyes in markers by running samples along a strip with an increasing solvent mixture as the mobile phase.
Paper chromatography is a technique used to separate colored chemicals or substances. It involves a stationary phase, usually a polar substance like water held in paper, and a mobile phase, generally a non-polar solvent mixture, that travels up the stationary phase through capillary action. Sample mixtures are spotted on the paper and separated as their components partition between the two phases based on differences in their affinity. Variations include ascending or descending based on solvent direction, and two-dimensional using two solvents rotated 90 degrees. The separation is analyzed by calculating Rf values, the ratio of compound travel distance to solvent front.
The versatile instrument is used to isolate unknown compounds from a HPTLC/TLC plate and transfer them into a mass spectrometer for identification or structure elucidation.
This document provides an overview of paper chromatography. It discusses the history and types of paper chromatography, including the principles of separation. It describes the practical requirements like the stationary and mobile phases used. It explains how to calculate Rf and Rx values and factors that can affect them. Various detection methods are outlined as well as sources of error. Finally, the applications and advantages of paper chromatography are highlighted.
This document provides information on paper chromatography, including its history, key terms, processes, techniques, factors that affect separation, and applications. Paper chromatography is an analytical technique used to separate mixtures based on differences in how compounds partition between a stationary phase, such as the paper, and a mobile phase, such as a solvent. Key points covered include the different types of paper chromatography, factors that influence separation like the solvent used, and how paper chromatography can be used to analyze organic compounds.
HPLC[ HIGH PERPROMANCE LIQUID CHROMATOGRAPHY OR HIGH PRESSURE LIQUID CHROMAT...Dr. Ravi Sankar
GASSCHROMATOGRAPHY[GC], ADVANCED STUDY OF THE FOLLOWING AND THEIR APPLICATIONS, INTRODUCTION, THEORY, COLUMN OPERATION,INSTRUMENTATION AND DETECTION,APPLICATIONS AND ADVANTAGES OF GC,PRINCIPLE OF SEPARATION IN GC, HOW GC MECHINE WORKS? COLUMN, DETECTORS.
BY P.RAVISANKAR, VIGNAN PHARMACY COLLEGE, VADLAMUDI, GUNTUR, A.P, INDIA.
Chromatographic techniques such as thin layer chromatography (TLC), high performance thin layer chromatography (HPTLC), column chromatography, and high performance liquid chromatography (HPLC) separate mixtures by distributing components between a stationary and mobile phase. HPLC uses high pressure to force a mobile liquid or gas phase through a column packed with solid particles. Components elute from the column at different rates and are detected and analyzed. Chromatographic techniques have applications in pharmaceutical analysis, environmental monitoring, food and flavor analysis, and forensics.
There are two theories that explain chromatography: plate theory and rate theory. Plate theory, developed in 1941, views the column as divided into theoretical plates where analytes completely equilibrate between the stationary and mobile phases. Rate theory, proposed in 1956, accounts for the dynamics of separation. Greater separation occurs with more theoretical plates and smaller plate height. The number of theoretical plates can be calculated using methods like half-height or USP, and depends on factors like column length, particle size, and retention time.
This document provides an overview of high performance liquid chromatography (HPLC). It discusses how HPLC refined traditional liquid chromatography by using smaller particle sizes, smaller column diameters, and high fluid pressures to provide enhanced separations over shorter periods of time. Key aspects of HPLC systems and processes are summarized, including the use of pumps to deliver mobile phases at high pressure through columns containing small stationary phase particles. Separation is achieved based on how sample components partition between the mobile and stationary phases. Various detectors are also outlined.
a type of an analyzer used in mass spectrometer. separates the ions based on mass to charge ratios. useful for the detection of ions present in the sample
The document discusses sample injection systems used in high performance liquid chromatography (HPLC). It describes the key components of sample injection valves, including a needle or syringe, metering device, sample loop, and valve. It also summarizes different types of injection systems, including manual injection, pull-to-fill autosamplers, push-to-fill autosamplers, and integral-loop autosamplers. Each type follows similar steps of loading the sample, injecting it, and washing the system.
High-performance thin layer chromatography (HPTLC) is an automated form of thin layer chromatography that provides higher efficiency and resolution than traditional TLC. HPTLC uses pre-coated plates with finer adsorbent particles, allowing for compact sample spots and bands. It can be used for both qualitative and quantitative analysis of natural products. HPTLC instrumentation includes automated sample applicators, development chambers, scanners, and software for detection and documentation.
This document discusses solid phase extraction (SPE), a popular sample preparation technique. SPE uses a solid stationary phase to isolate and concentrate analytes from liquid samples. It reduces interference levels and minimizes final sample volumes. The degree of analyte enrichment depends on the selectivity and strength of interaction between the analyte and bonded phase. Common retention mechanisms include non-polar, polar, ionic, and hydrogen bonding interactions. SPE provides flexibility, longer column lifetimes, better contaminant removal, recovery, reproducibility and sensitivity compared to liquid-liquid extraction.
Chromatography : A seperation techniqueSHIVANEE VYAS
Chromatography is a method of seperating mixture of components into individual components through equlibrium distribution between two phases.
Each chromatographic method essentially consists of 2 phases a staionary phase and a mobile phase.
Stationary phase : solid or liquid
Mobile phase : liquid or gas
The document discusses the history and development of thin layer chromatography (TLC). It describes how TLC works and the basic components and steps involved, including the stationary and mobile phases, developing tanks, detection methods, and applications. Some key advantages of TLC are that it is simple, rapid, requires small sample sizes, and uses less solvent than other chromatography methods.
HPTLC, or high-performance thin-layer chromatography, is a sophisticated form of thin-layer chromatography that allows for quantitative analysis. It works on the same principles as TLC by separating components via adsorption as the mobile phase moves up the stationary phase. HPTLC provides advantages like visual chromatograms, simplicity, ability to handle multiple samples simultaneously, and low costs. It involves steps like sample preparation, selecting and pre-treating plates, applying samples, developing the plate, detecting and quantifying spots through densitometry, and documenting results. HPTLC is used in applications like pharmaceutical quality control, food analysis, clinical testing, and industrial process development.
NMR spectroscopy is an analytical technique that uses the magnetic properties of certain nuclei, such as 1H and 13C, to characterize organic molecules. It was independently developed in the 1940s-1950s by groups at Harvard and Stanford, with Nobel Prizes awarded. There are two main types - 1H NMR studies hydrogen atoms and 13C NMR studies carbon atoms. The instrument uses a strong magnet to align nuclear spins, radio waves to excite them, and detectors to measure the radiofrequency energy emitted as the spins relax. NMR provides information about a molecule's structure through analysis of peak positions in its spectrum.
This document discusses the principles and instrumentation of high performance thin layer chromatography coupled with mass spectrometry (HPTLC-MS). HPTLC-MS combines the separation capabilities of HPTLC with the identification abilities of mass spectrometry. It works by separating compounds via HPTLC and then using an interface to extract zones of interest from the HPTLC plate and transfer them to a mass spectrometer for analysis. This allows for structural confirmation and elucidation of targeted analytes. Some advantages of HPTLC-MS are that it is cost-effective, allows identification of unknown substances, and is useful for applications like drug analysis, metabolism studies, and characterization of impurities.
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.
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.
This document discusses different types of chromatography techniques and the pumps used in each. It covers high performance liquid chromatography (HPLC), ion-exchange chromatography, and size-exclusion chromatography. For HPLC, it describes reciprocating piston pumps that are able to deliver precise, pulse-free flow at high pressures up to 10,000 psi. For ion-exchange chromatography, it mentions pumps must provide pulse-free flow for sensitive detectors and single piston pumps are commonly used. Size-exclusion chromatography utilizes small volume reciprocating pumps for accurately controlled flow rates at pressures up to 7,250 psi.
Hii..
in which slide we are involving what is Gas chromatography there History, Theory & principle, Introduction, Phases, Types, Instrumentation, Application etc.
Paper chromatography is a method used to separate components of a mixture based on how they partition between a stationary phase, typically cellulose paper, and a mobile phase, typically a solvent. Key aspects of paper chromatography include choosing an appropriate paper and solvent based on the sample's properties, applying the sample as a spot on the paper, developing the chromatogram by allowing the solvent to migrate up the paper via capillary action, and detecting separated components using physical or chemical methods. Results are represented by each component's retention factor (Rf) value, which indicates its relative migration distance compared to the solvent front. Variations in development technique include ascending, descending, and two-dimensional paper chromatography. Paper chromatography has wide applications in chemistry, biology
This document provides an overview of chromatography including its history, principles, types, and applications. It discusses how chromatography works and key terms. Chromatography is introduced as a laboratory technique used to separate mixtures based on differences in how components partition between a stationary and mobile phase. The document summarizes different chromatography methods including paper chromatography, thin layer chromatography, gas chromatography, high performance liquid chromatography, and more. It provides examples of chromatography principles and procedures.
Chromatographic techniques such as thin layer chromatography (TLC), high performance thin layer chromatography (HPTLC), column chromatography, and high performance liquid chromatography (HPLC) separate mixtures by distributing components between a stationary and mobile phase. HPLC uses high pressure to force a mobile liquid or gas phase through a column packed with solid particles. Components elute from the column at different rates and are detected and analyzed. Chromatographic techniques have applications in pharmaceutical analysis, environmental monitoring, food and flavor analysis, and forensics.
There are two theories that explain chromatography: plate theory and rate theory. Plate theory, developed in 1941, views the column as divided into theoretical plates where analytes completely equilibrate between the stationary and mobile phases. Rate theory, proposed in 1956, accounts for the dynamics of separation. Greater separation occurs with more theoretical plates and smaller plate height. The number of theoretical plates can be calculated using methods like half-height or USP, and depends on factors like column length, particle size, and retention time.
This document provides an overview of high performance liquid chromatography (HPLC). It discusses how HPLC refined traditional liquid chromatography by using smaller particle sizes, smaller column diameters, and high fluid pressures to provide enhanced separations over shorter periods of time. Key aspects of HPLC systems and processes are summarized, including the use of pumps to deliver mobile phases at high pressure through columns containing small stationary phase particles. Separation is achieved based on how sample components partition between the mobile and stationary phases. Various detectors are also outlined.
a type of an analyzer used in mass spectrometer. separates the ions based on mass to charge ratios. useful for the detection of ions present in the sample
The document discusses sample injection systems used in high performance liquid chromatography (HPLC). It describes the key components of sample injection valves, including a needle or syringe, metering device, sample loop, and valve. It also summarizes different types of injection systems, including manual injection, pull-to-fill autosamplers, push-to-fill autosamplers, and integral-loop autosamplers. Each type follows similar steps of loading the sample, injecting it, and washing the system.
High-performance thin layer chromatography (HPTLC) is an automated form of thin layer chromatography that provides higher efficiency and resolution than traditional TLC. HPTLC uses pre-coated plates with finer adsorbent particles, allowing for compact sample spots and bands. It can be used for both qualitative and quantitative analysis of natural products. HPTLC instrumentation includes automated sample applicators, development chambers, scanners, and software for detection and documentation.
This document discusses solid phase extraction (SPE), a popular sample preparation technique. SPE uses a solid stationary phase to isolate and concentrate analytes from liquid samples. It reduces interference levels and minimizes final sample volumes. The degree of analyte enrichment depends on the selectivity and strength of interaction between the analyte and bonded phase. Common retention mechanisms include non-polar, polar, ionic, and hydrogen bonding interactions. SPE provides flexibility, longer column lifetimes, better contaminant removal, recovery, reproducibility and sensitivity compared to liquid-liquid extraction.
Chromatography : A seperation techniqueSHIVANEE VYAS
Chromatography is a method of seperating mixture of components into individual components through equlibrium distribution between two phases.
Each chromatographic method essentially consists of 2 phases a staionary phase and a mobile phase.
Stationary phase : solid or liquid
Mobile phase : liquid or gas
The document discusses the history and development of thin layer chromatography (TLC). It describes how TLC works and the basic components and steps involved, including the stationary and mobile phases, developing tanks, detection methods, and applications. Some key advantages of TLC are that it is simple, rapid, requires small sample sizes, and uses less solvent than other chromatography methods.
HPTLC, or high-performance thin-layer chromatography, is a sophisticated form of thin-layer chromatography that allows for quantitative analysis. It works on the same principles as TLC by separating components via adsorption as the mobile phase moves up the stationary phase. HPTLC provides advantages like visual chromatograms, simplicity, ability to handle multiple samples simultaneously, and low costs. It involves steps like sample preparation, selecting and pre-treating plates, applying samples, developing the plate, detecting and quantifying spots through densitometry, and documenting results. HPTLC is used in applications like pharmaceutical quality control, food analysis, clinical testing, and industrial process development.
NMR spectroscopy is an analytical technique that uses the magnetic properties of certain nuclei, such as 1H and 13C, to characterize organic molecules. It was independently developed in the 1940s-1950s by groups at Harvard and Stanford, with Nobel Prizes awarded. There are two main types - 1H NMR studies hydrogen atoms and 13C NMR studies carbon atoms. The instrument uses a strong magnet to align nuclear spins, radio waves to excite them, and detectors to measure the radiofrequency energy emitted as the spins relax. NMR provides information about a molecule's structure through analysis of peak positions in its spectrum.
This document discusses the principles and instrumentation of high performance thin layer chromatography coupled with mass spectrometry (HPTLC-MS). HPTLC-MS combines the separation capabilities of HPTLC with the identification abilities of mass spectrometry. It works by separating compounds via HPTLC and then using an interface to extract zones of interest from the HPTLC plate and transfer them to a mass spectrometer for analysis. This allows for structural confirmation and elucidation of targeted analytes. Some advantages of HPTLC-MS are that it is cost-effective, allows identification of unknown substances, and is useful for applications like drug analysis, metabolism studies, and characterization of impurities.
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.
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.
This document discusses different types of chromatography techniques and the pumps used in each. It covers high performance liquid chromatography (HPLC), ion-exchange chromatography, and size-exclusion chromatography. For HPLC, it describes reciprocating piston pumps that are able to deliver precise, pulse-free flow at high pressures up to 10,000 psi. For ion-exchange chromatography, it mentions pumps must provide pulse-free flow for sensitive detectors and single piston pumps are commonly used. Size-exclusion chromatography utilizes small volume reciprocating pumps for accurately controlled flow rates at pressures up to 7,250 psi.
Hii..
in which slide we are involving what is Gas chromatography there History, Theory & principle, Introduction, Phases, Types, Instrumentation, Application etc.
Paper chromatography is a method used to separate components of a mixture based on how they partition between a stationary phase, typically cellulose paper, and a mobile phase, typically a solvent. Key aspects of paper chromatography include choosing an appropriate paper and solvent based on the sample's properties, applying the sample as a spot on the paper, developing the chromatogram by allowing the solvent to migrate up the paper via capillary action, and detecting separated components using physical or chemical methods. Results are represented by each component's retention factor (Rf) value, which indicates its relative migration distance compared to the solvent front. Variations in development technique include ascending, descending, and two-dimensional paper chromatography. Paper chromatography has wide applications in chemistry, biology
This document provides an overview of chromatography including its history, principles, types, and applications. It discusses how chromatography works and key terms. Chromatography is introduced as a laboratory technique used to separate mixtures based on differences in how components partition between a stationary and mobile phase. The document summarizes different chromatography methods including paper chromatography, thin layer chromatography, gas chromatography, high performance liquid chromatography, and more. It provides examples of chromatography principles and procedures.
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.
Chromatography is a family of analytical chemistry techniques used to separate mixtures based on differences in how components interact with two phases - a stationary phase and a mobile phase. The document discusses the history and definition of chromatography, as well as several types including liquid chromatography, gas chromatography, paper chromatography, thin layer chromatography. It provides examples of chromatography uses in fields like petroleum engineering, forensics, and environmental testing. The key aspects and processes of each technique are described.
Introduction to chromatography and its applications 2Kalsoom Mohammed
Chromatography is a technique used to separate mixtures based on differences in how components interact with stationary and mobile phases. The document defines chromatography and describes its history, principles, commonly used terms, types including adsorption (gas chromatography, thin layer chromatography, column chromatography, ion exchange chromatography, HPLC) and partition (paper chromatography, gas chromatography), working, detectors, visualization, applications and references. Chromatography is widely used in fields like pharmaceuticals, food, forensics and more to analyze and purify chemical mixtures.
Chromatography is 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.
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.
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.
Chromatography is a technique used to separate mixtures by distributing components between two phases - a stationary phase and a mobile phase. Paper chromatography uses a cellulose paper as the stationary phase and a solvent as the mobile phase. Components separate based on how strongly they interact with each phase. The separation can be visualized by detecting spots, and identified using the retention factor (Rf) which characterizes how far each component travels up the paper relative to the solvent front. Paper chromatography is used to analyze, identify, purify, and quantify mixtures in applications like pharmaceutical analysis, forensics, and food and drug testing.
Chromatography is a separation technique that involves distributing components of a mixture between two phases, one stationary and one mobile. Paper chromatography is a type of chromatography where the stationary phase is held in the fibers of filter paper and the mobile phase is a solvent that travels up or down the paper. Key aspects of paper chromatography include choosing an appropriate solvent system based on the compounds' polarity, spotting samples on the paper, developing the paper in the solvent, and visualizing the separated components using chemical or physical detection methods. Paper chromatography is useful for qualitative analysis of mixtures in various fields.
The document discusses different types of chromatography techniques used to separate mixtures. It describes chromatography as a collective term for laboratory methods that separate mixtures by dissolving them in a mobile phase that carries the mixture through a stationary phase. The key types discussed are paper chromatography, thin layer chromatography, gas chromatography, column chromatography, and high performance liquid chromatography. HPLC uses very small column packing and high pressure to improve separation speed over other chromatography methods.
Chromatography is a physical separation technique that separates the components of a mixture based on their interactions with two phases - a stationary phase and a mobile phase. Mikhail Tswett discovered chromatography in 1906 while separating plant pigments. Chromatography has many applications and has contributed to 12 Nobel prizes between 1937-1972. There are several types of chromatography classified by the mobile phase (liquid, gas), stationary phase (thin layer, paper, column), or separation force (adsorption, partition, ion exchange, gel filtration, affinity). High performance liquid chromatography uses high pressure to allow for faster separation over minutes versus hours with other techniques. Chromatography techniques like thin layer chromatography, gas chromatography, and HPLC are used in various
Chromatography is a technique used to separate mixtures based on how their constituents partition between a stationary and mobile phase. In paper chromatography, the stationary phase is held in the pores of filter paper while the mobile phase moves over the paper. Different compounds travel at different speeds depending on how they partition between the water in the stationary phase and the mobile solvent, allowing separation. Column chromatography uses a column filled with a stationary phase like silica gel and a mobile phase solvent that carries compounds through the column at different rates based on their interactions with the stationary phase. Thin layer chromatography plates have a thin stationary phase coating and compounds are separated as they move up the plate with the mobile phase solvent, appearing as separated spots. These techniques are used
This document provides an overview of chromatographic techniques. It begins with definitions and a brief history, then covers principles, applications, classification, specific techniques (e.g. gas chromatography, liquid chromatography), terms, and properties of stationary phases. The document presents chromatographic methods and their use in separating mixtures like drugs, proteins, and other compounds. It concludes that supercritical fluid chromatography falls between HPLC and GC in performance for applications in pharmaceutical and bioanalytical analysis.
This document provides an overview of chromatographic techniques. It begins with definitions and a brief history, explaining that chromatography separates mixtures based on how components partition between a mobile and stationary phase. The document then covers key terms, classifications of chromatography by mechanism and phase, and specific techniques like gas-liquid chromatography, solid-liquid chromatography, and thin layer chromatography. It discusses important properties of liquid stationary phases and concludes that chromatography techniques like supercritical fluid chromatography fall between HPLC and GC in terms of separation capabilities.
Chromatography is a technique used to separate chemical mixtures by exploiting differences in how components interact with stationary and mobile phases. It was first developed in 1900 to separate plant pigments. The key is separation of components to simplify analysis of unknown substances. Chromatography can be classified based on the physical means of contact between phases, the type of mobile and stationary phases used, and the type of interactions that occur. The efficiency of separation depends on column resolution, which is improved by increasing the difference in retention times and decreasing peak widths. Migration rates are determined by distribution constants between phases and affect retention times.
This document provides an introduction to chromatography, including its history and essential features. It discusses the basic components and process of chromatography, including the stationary and mobile phases. It also describes different types of chromatography techniques based on the stationary phase, such as partition chromatography, adsorption chromatography, ion exchange chromatography, molecular exclusion chromatography, and affinity chromatography. Finally, it discusses applications of chromatography in qualitative analysis, quantitative analysis, and preparative purposes.
Chromatography is a technique used to separate mixtures by distributing components between two phases, stationary and mobile. The mixture is dissolved in a mobile phase that carries it through a column containing a stationary phase. Components travel at different rates based on how they partition between the phases, allowing separation. Common chromatography methods include gas chromatography, liquid chromatography, and thin layer chromatography. Chromatography has applications in identifying unknown substances like drugs, proteins, and plant pigments. It was first developed in 1903 and continues to be an important analytical technique.
Chromatography is a technique used to separate mixtures into individual components. It works by exploiting differences in how fast components migrate and adhere to stationary and mobile phases. There are several types including paper chromatography, thin layer chromatography, gas chromatography and liquid chromatography. Paper chromatography involves using a stationary phase like filter paper and a mobile phase like water or solvent to separate ink components by capillary action. Thin layer chromatography uses a thin silica plate as the stationary phase and a solvent as the mobile phase to separate components into spots visualized by color reagents. Chromatography is used in various applications such as forensic analysis and identifying biological molecules.
4. DEFINATION
The technique of separating
the components of a mixture is
achieved by the differential
movement of individual
components through a
stationary phase under the
influence of a mobile phase.
Chromatography is widely
used for separation ,
purification , identification
and characterisation of the
components of a mixture
wether coloured or colourless.
5. Principles of chromatography
Chromatography is a physical process.
Any chromatography system is composed of mainly
three things:
1) Stationary phase
2) Mobile phase
3) Mixture to be separated
6. Chromatography is an dynamic process in which
mobile phase moves in definite direction.
In chromatography process we can only control
stationary &mobile phase as mixture is the problem we
have to deal with.
7. TERMINOLOGIES
Differential: showing a difference.
Affinity: natural force of attraction between the
substances.
Adsorption :phenomenon of higher concentration of
molecular species(liquid or gas) on the surface of solid in
bulk.
Adsorbent: the substance’s surface on which adsorption
took place.
Adsorbate : substance which get adsorbed on adsorbent.
Stationary phase : part of apparatus that does not move
with the sample.
Mobile phase : gas or liquid that carries the sample with
it.
8. WORKING OF CHROMATOGRAPHY
The substance which has to be separated has many
components, some of them has affinity to mobile
phase & some has affinity to stationary phase.
The substance having affinity to mobile phase get
dissolved in it and travels large distance it.
While substance having affinity to stationary phase get
adsorbed on it and travels very short distance.
9. Uses of chromatography
Analyze: examine of mixture, it’s components & there
relation with one another.
Identify : determine the identy of mixture or
components based on known components.
Purify: separates components in order to isolate
components for further studies.
Quantify : dertimens the amount of substances
present in the sample.
15. STATIONARY PHASE
The stationary phase or adsorbent in column
chromatography is a solid.
The most common stationary phase for column
chromatography is silica gel, followed
by alumina. Cellulose powder has often been used in the
past.
The stationary phases are usually finely ground powders
or gels and/or are micro porous for an increased surface.
16. MOBILE PHASE
•The mobile phase or eluent is either a
pure solvent or a mixture of different
solvents.
• The eluent has also been chosen so that
the different compounds can be separated
effectively
17.
18. Column chromatography
Column chromatography in chemistry is a method used
to purify individual chemical compounds from mixtures of
compounds.
The main advantage of column chromatography is the
relatively low cost and disposability of the stationary
phase used in the process. The latter prevents cross-
contamination and stationary phase degradation due to
recycling
We can’t separate different amino acid’s &sugars. We
can’t also do quantitative analyses accurately.
20. 6 beakers or jars
6 covers or lids
Distilled H2O
Isopropanol
Graduated cylinder
6 strips of filter paper
Different colors of Sharpie
pens
Pencil
Ruler
Scissors
Tape
Materials List
21. STATIONARY PHASE
•The stationary phase or adsorbent in paper
chromatography is a liquid
Paper is made of cellulose fibres, and cellulose is a
polymer of the simple sugar, glucose.
The key point about cellulose is that the polymer
chains have –OH groups sticking out all around
them. To that extent, it presents the same sort of
surface as silica gel or alumina in thin layer
chromatography
cellulose fibres attract water vapour from
the atmosphere as well as any water that
was present when the paper was made.
You can therefore think of paper as being
cellulose fibres with a very thin layer of
water molecules bound to the surface.
22. MOBILE PHASE
The mobile phase used in paper chromatography is liquid
. non-polar solvent such as hexane, Acetone,etc, Polar
solvents water, alcohols,etc as the solvent
The solvent has also been chosen so that the different
compounds can be separated effectively
23. PRINCIPLE OF PAPER
CHROMATOGRAPHY
Capillary Action – the movement of liquid within the spaces of a
porous material due to the forces of adhesion, cohesion, and surface
tension. The liquid is able to move up the filter paper because its
attraction to itself is stronger than the force of gravity.
Solubility – the degree to which a material (solute) dissolves into a
solvent. Solutes dissolve into solvents that have similar properties.
(Like dissolves like) This allows different solutes to be separated by
different combinations of solvents.
It can be used to separate different
amino acids & sugars but quantitative
analyses is not accurately.
24. THIN LAYER CHROMATOGRAPHY
Here the mobile phase is a liquid, flowing past
a thin layer of powder on a solid support by
CAPILLARY ACTION; a variation of column
chromatography.
Substances that are less attracted to
the solid or are more soluble in the
liquid move faster. And so move
further up the plate by the time that
the process has been stopped by
taking the plate out of the liqiud. -
larger Rf
26. . Detection
If the spots are not colored and can’t be seen by the
eye, use:
• UV lamp for UV-active compounds; most aromatics
are UV-active
• If compounds are not UV-active, use an iodine .
27. Rf value(retention factor)
The ratio of distance travelled by the component (from
origin) compared with the distance travelled by the
solvent front (from origin) is called the Rf value.
Rf = Migration distance of a substance
Migration distance of solvent front
Origin Line
Solvent Front Line
Distance traveled
by solvent
Distance traveled by spot
28. 28
••
The ratio of distance travelled by the component (from origin) compared
with the distance travelled by the solvent front (from origin) is called the Rf
value.
Solvent front
x
a
b
c
Rf of = a/x
Rf of = b/x
Rf of = c/x
29. Rf Value
•The Rf value is not
informative
•What affects the Rf value?
Temperature
Solvent
Thickness and amount
of spot
31. ?
GAS CHROMATOGRAPNY
1) In gas chromatography the moving
phase is a gas and the stationary phase
is either liquid or solid.
2) The technique is suitable for
separation of materials which are
volatile without decomposition
32. It can be used for qualitative as well
as quantitative analyses of
substances.
It can’t be used to separate non-
volatile substances.
33. HIGH PRESSURE LIQUID
CHROMATOGRAPHY
Different analytes have different equilibria between the
mobile phase and stationary phase
Equilibrium is dynamic; thus we can view it as a given
analyte molecule spending a fraction of time dissolved in
the mobile phase
Since different solutes gave different fractions, a separation
of the analytes occur as they are pushed through the
column by the mobile phase
34. Uses for Chromatography
Real-life examples of uses for chromatography:
• Pharmaceutical Company – determine amount of
each chemical found in new product
• Hospital – detect blood or alcohol levels in a
patient’s blood stream
• Law Enforcement – to compare a sample found at
a crime scene to samples from suspects, Like
Explosion residue, Arson cases Poisons in viscera.
• Environmental Agency – determine the level of
pollutants in the water supply
• Manufacturing Plant – to purify a chemical
needed to make a product
• Dyes –Identifying coloring dyes used in different
food products