Ppt on column chromatography applications

B y: Nuriya Khan.  Chromatography is used for the separation, identification and measurement of the chemical components in mixtures.  There are a.

of a chromatographic separation Chromatographical Methods  Liquid Chromatography  Gas Chromatography  Ion Exchange  Gel permeation Chromatography  The diagram on the following slide illustrates separation of solutes in a solution by column chromatography. The stationary phase is an inert solid./, vertically so that the capillary empties itself and carefully to avoid damage to the layer. Easy application of samples is allowed with a spotting guide.  When the solvent is completely evaporated (approx./


LECTURE 4: CHROMATOGRAPHY Content: - Chromatographic separation - classifying analytical separations; column chromatography, planar chromatography - gas.

Content: - Chromatographic separation - classifying analytical separations; column chromatography, planar chromatography - gas chromatography; introduction, application - hplc; introduction, application Simple separation VS Chromatographic separation Principle of simple separation (exp partitioning between phases): -The separation occurs only one direction -Increase efficiency by using fresh extracting phase. The principle of chromatographic /


Liquid Chromatography (GC) Hashim Baharin Department of Chemistry, Faculty of Science Universiti Teknologi Malaysia 81310 Skudai, Johor, Malaysia.

for a column chromatographic separation.To determine the appropriate conditions for a column chromatographic separation. To monitor column chromatographyTo monitor column chromatography. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) Evolved from preparative column chromatography. Performance / of sample components, ion-pair reagents to neutralize charged solutes and make them more lipophilic Applications of HPLC Reverse Phase Proteins,- including cereal proteins. Water soluble and fat soluble vitamins. /


Prof. aza 1 Gas chromatography- mass spectrometry (Additional Reading) prof. aza.

few select ions and return values as ratios. 21prof. aza References Eiceman, G.A. (2000). Gas Chromatography. In R.A. Meyers (Ed.), Encyclopedia of Analytical Chemistry: Applications, Theory, and Instrumentation, pp. 10627. Chichester: Wiley. ISBN 0-471-97670-9 ISBN 0-471-97670/. The stationary phase is usually contained in a tube of some sort. This tube is referred to as a column. Columns can be glass or stainless steel of various dimensions. 27prof. aza The mixture of compounds in the mobile phase /


HIGH PERFORMANCE LIQUID CHROMATOGRAPHY PRESENTED BY P.SOBHITHA RANI.

the technique of liquid column chromatography because of the development of HPLC by Kirkland and Huber. They proposed high pressure systems capable of operating at pressures up to 3000psi. In HPLC, small diameter columns(1-3mm) with / detector for the fluorimetric analysis of carbohydrates in body fluids using Ce (III) flourescence has also been reported. APPLICATIONS: HPLC is one of the most widely applied analytical separation techniques. Pharmaceutical: Tablet dissolution of pharmaceutical dosages. Shelf/


High Performance/Pressure Liquid Chromatography (HPLC)

for High Performance/ Pressure Liquid Chromatography. It is a kind of sophisticated and mechanized column chromatography. It uses high pressure to drive the mobile phase through the (column of) stationary phase. Chromatography: Method Chromatograph: Machine Chromatographer/ of this type of detectors are their high sensitivity, selectivity, simplicity, convenience and widespread applicability. These devices are based upon amperometry, polarography, coulometry, and conductometry. Analytes are usually readily/


FLASH CHROMATOGRAPHY FLASH CHROMATOGRAPHY. PRESENTED BY :- MISS.PRACHITEE PRAKASH AYARE. M.PHARM FIRST YEAR. (QUALITY ASSURANCE) UNDER THE GUIDANCE OF.

of the sample can be separated (0.5- 2g)  Fast ( 1o to 15 minutes)  Cost efficient  If high resolution is required, flash chromatography is carried out before HPLC to avoid contamination of the expensive columns. 25 APPLICATIONS Flash Chromatography has various applications in following fields  SYNTHETIC CHEMISTRY It is used as a tool to monitor the reaction progress and to isolate and identify a mixture/


Optimisation and control of chromatography

bed is simulated. Complex mixed discrete and continuous dynamics SMB chromatography: process dynamics Continuous flows and discrete switchings Axial profile builds up during start-up Same profile in different columns in cyclic steady state Periodic output concentrations The VARICOL process Variable length column process (NovaSEP 2000) Periodic but asynchronous switching of the ports Industrial applications of SMB I Petro-chemicals Universal Oil Products (USA/


SIZE EXCLUSION CHROMATOGRAPHY

Equipment for running size exclusion chromatography. The buffer is pumped through the column by a computer controlled device Illustrative description of separation in SEC. (From Introduction to Modern Liquid Chromatography, 2nd edition by L. Snyder and J. J. Kirkland, © 1979 by John Wiley & Sons, Inc. ) Physical Pharmacy 2 KBA Molecular weight determination Physical Pharmacy 2 4/1/2017 Applications Fractionation Desalting Concentration Molecular weight/


Gas Chromatography.

& Zlatkis, 1967, “The Practice of Gas Chromatography) stationary bed fluid History of 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 Applications gas Compound must exist as a ____ at a temperature that can be produced by the GC and withstood by the column (up to 450°C) Alcohols in/


Bioseparation Chapter 9

molecules as part of a manufacturing process, it is referred to as preparative chromatography. Some of the applications in biotechnology: 1. Biopharmaceutical production 2. Biopharmaceutical and biomedical analysis 3. Environmental analysis 4. Foods and nutraceuticals production 5. Diagnostics 6. Process monitoring Chromatography system A chromatographic separation system consists of a column, mobile phase reservoir/s, pump/s, sample injector, detector/s and sometimes a/


Liquid Chromatography 2 Lecture Date: April 14 th, 2008.

flow rates with lower backpressure –Stable over a wide pH range Three Dionex monolithic columns compared with a polymer bead (particle) column Figures from Dionex, Inc. application note, www.dionex.comwww.dionex.com Two different flow rates on a monolithic columns (viper venom, a complex biological mixture) Two-Dimensional Liquid Chromatography (2D-LC) ● 2D LC: two LC experiments run back-to-back, with the effluent/


Gas Chromatography.

The comparison of retention times is what gives GC its analytical usefulness. Gas chromatography and column chromatography Gas chromatography is in principle similar to column chromatography, but has several notable differences: The process of separating the compounds in a/ test could statistically suggest the identity of the substance, this could lead to false positive identification. Gas Chromatography Application In general, substances that vaporize below ca. 300 °C (and therefore are stable up to that /


Chromatography.

“artifactual” peaks Shortest possible analysis times Resolution and Retention time are key Instrumentation (1) Origins Preparative column chromatography (Glass columns) Internal diameter: 1 - 5 cm Length: 50 - 500cm Particle size: 150-200mm (large) / removed for further analysis Applications Wide range including: Drugs, foods, pesticides, polymers, surfactants…. Advanced Techniques Immunoaffinity techniques Tandem techniques Advanced Techniques (1) Immunoaffinity chromatography Most often for solid-phase/


Downstream Processing in Biopharmaceutical Manufacturing Harvest and Clarification Tangential Flow Filtration (UF/DF) Low Pressure Liquid Column Chromatography.

Tank 6,000L Eluate Hold Tank 6,000L Filter Chromatography Skid Protein A Chromatography Column Chromatography Skid Column Eluate Hold Tank 20,000L Eluate Hold Tank 20,000L Hydrophobic Interaction Chromatography (HIC) Eluate Hold Tank 20,000L Eluate Hold / of the membrane. Contaminants smaller than the specified pore size pass through the membrane. Used for critical applications such as sterilizing and final filtration. Normal Flow Filtration Depth Filtration: Equipment Depth Filtration: Cells and Cellular/


HPLC Instrumentation and Applications. Other names for HPLC 1- High Speed Liquid Chromatography - As the separation is completed within few minutes. 2-

Applications Other names for HPLC 1- High Speed Liquid Chromatography - As the separation is completed within few minutes. 2- High Performance Liquid Chromatography (HPLC) 3- High Resolution Liquid Chromatography (HRLC) High performance is the result of many factors: - Smaller particles of the stationary phase, uniform pore size, high pressure column/mixture of water and methanol or acetonitrile). 2- Size exclusion chromatography. The column is packed with material having controlled pore sizes and the sample/


Chapter 5c : LIQUID CHROMATOGRAPHY Department of Chemistry, Faculty of Science Universiti Teknologi Malaysia ___________________________ Analytical Chemistry.

a column chromatographic separation.To determine the appropriate conditions for a column chromatographic separation. To monitor column chromatographyTo monitor column chromatography. HIGH PERFORMANCE LIQUID CHROMATOGRAPHY (HPLC) Evolved from preparative column chromatography. Performance/species and large molecules. Analytical or preparative (highly purified compound in small or large quantities). Applications include analysis of sugars, pesticide residues, organic acids, lipids, amino acids, toxins, and /


Lecture I. 1- Chromatographic methods A. Braithwait, E.J. Smith (1995) 2- Modern thin layer chromatography (chromatographic science services vol, 52)

equipment and the operational procedures A- Column chromatography - The stationary phase is packed in tube and mobile phase pass through it by gravity or pressure -Column chromatography - HPLC - GC - SEC B- Planar chromatography The stationary phase is solid coated onto/ leading to tailing 1. strong interaction between solute and stationary phase 2. application of excessive amount of the sample to the column 3. poor column packing 4. improper selection of mobile phase  occurs when the interaction between/


Trends in development of stationary phases in chromatography Petr Solich Charles University Czech Republic Separation Techniques, San Francisco, USA -

mellitus, pancreatitis) - clinical monitoring of elderly patients (level of antioxidants) - monitoring of patients with nephorological infections Bioanalytical applications of monolithic columns Determination of retinoic acids (RA) Most important RA are 13-cis RA, all-trans RA For treatment of hemato-/ 80 points/s  special optical cell (500 nl, 10 mm) Ultra High Performance Liquid Chromatography (UHPLC) Time of analysis: 16.3 min Solvent consumption: 11.4 ml Time of analysis: 2.3 min Solvent consumption /


HIGH PERFORMANCE LIQUID CHROMATOGRAPHY BY K RAKESH GUPTA 1.

destructive 3 HPLC  HPLC- It was originally referred to as High Pressure Liquid Chromatography since high pressure is applied using a pumping system to the column.  This pressure works by forcing the mobile phase through, at much higher rate/ The diffusion current recorded is directly proportional to the concentration of the compound recorded. DISADVANTAGE: This detector is applicable only when the functional groups present in the sample can be either oxidized or reduced. ADVANTAGE: Highly sensitive /


Chapter 32 Gas Chromatography. In gas chromatography, the components of a vaporized sample are separated by being distributed between a mobile gaseous.

fall within a suitable range. To have a reasonable residence time on the column, an analyte must show some degree of compatibility (solubility) with the stationary phase especially with respect to the polarities of the analyte and the immobilized liquid. 32 C Applications of gas-liquid chromatography It is applicable to species that are appreciably volatile and thermally stable at temperatures up to a/


Gas Chromatography. Inventers Russian MS Tswett (1903) – invented Chromatography German Fritz Prior (1947) – invented Gas solid chromatography Archer.

a limited use due to long retention of active or polar molecules and severe tailing of elution peaks –Real application is for specific low MW compounds Gas-Liquid Chromatography (GLC) –Has a thin layer of a liquid stationary phase immobilised inside the column and the compounds partition between the gaseous mobile phase and the liquid phase. –Widely adopted method and is now known/


BY G.SHRAVANI 170213884010 ION EXCHANGE AND GEL PERMEATI ON CHROMATOGRAPHY 1G.SHRAVANI.

of ion exchangers. Isoelectric focusing can be used to identify suitable ion-exchanger type 20G.SHRAVANI Applications Determination of sodium and potasium in the mixture: column eluted with 0.1M HCL flow rate -0.6ml/sqmin Radiochemistry G.SHRAVANI21 Gel Permeation GEL FILTRATION Chromatography 22G.SHRAVANI DEFINATION Gel chromatography is a technique in which fractionation is based upon the molecular size & shape of the/


Principles of chromatography. ChromatographyChromatography’ term - Mikhail Tswett (1906) Chroma = color, graphein = written Each substance  Different.

binding site in the stationary phase appears Molecule will not elute from the column until a solution of varying pH or ionic strength is passed through it Thus, separation is highly selective Applications Purification of biological materials Separation of charged compounds like the peptides, amino acids, proteins, etc. Affinity chromatography Stationery : Immobilized molecule, an antibody to some specific protein on a solid matrix/


COLUMN CHROMATOGRAPHY. Presented By – Mr. Shaise Jacob Faculty Nirmala College of Pharmacy Muvattupuzha, Kerala India E mail –

improved by decreasing the particle size of the adsorbent. 3. Activity of the adsorbent 4.Temperature of the column: The speed of the elution increases at higher temperatures. 5.Packing of the column 6.Quality of solvents: solvents having low viscosities is giving better results. COLUMN CHROMATOGRAPHY APPLICATIONS ► Separation of mixture of compounds ► Purification process ► Isolation of active constituents ► Estimation of drugs in formulation ► Isolation/


Gas Chromatography. Presented By - Mr. Shaise Jacob Faculty Nirmala College of Pharmacy Muvattupuzha, Kerala India –

. (1950) The Next Generation in Gas Chromatography How a Gas Chromatography Machine Works –First, a vaporized sample is injected onto the chromatographic column. –Second, the sample moves through the column through the flow of inert gas. –Third/Hexane0.12 Argon0.12 Methanol0.13 Nitrogen0.17 Helium1.00 Hydrogen1.28 Advantages of Katharometer  Linearity is good  Applicable to most compounds  Non destructive  Simple & inexpensive Disadvantages Low sensitivity Affected by fluctuations in temperature and flow/


1. Gas Chromatography Mass Spectrometry (GC-MS) Presented by :Ayesha Abdul Ghafoor Student ID : MS (I) (Analytical Chemistry section) Session : 2011-13.

Instrument Fig 1:The insides of the GC-MS, with the column of the gas chromatograph in the oven on the right. 13 GAS CHROMATOGRAPHY Gas chromatography leads to Separation of volatile organic compounds Separation occurs as a result/ two other questions are addressed: Is the identification plausible? Why is it present in a given sample? 51 Applications of GC-MS Petrochemical and hydrocarbons analysis Geochemical research Forensic (arson, explosives, drugs, unknowns) Environmental analysis Pesticide analysis/


Chromatography Copyright © 28/05/2013 by Omer Bayazeid. All rights reserved.

Traditional column chromatography is characterized by addition of mobile phase under atmospheric pressure and the stationary phase is packed in a glass column. Open Column Chromatography (Traditional column chromatography) Detection: On-column detection/Gas Liquid Chromatography Schematic Diagram of Gas Chromatography 1)Flame Ionization Detector (Nanogram - ng). 2) Thermal Conductivity Detector. 3) Electron Capture Detector. Schematic Diagram of Gas Chromatography DETECTORS Gas Chromatography Application 1)/


INTRODUCTION OF GAS CHROMATOGRAPHY

GC TYPE OF GC COLUMNS OF GC:- packed column open(capillary) column COLUMN SELECTION PARAMETERS GC BLOCK DIAGRAM SAMPLE FOR GC INSTRUMENTATION DETECTORS CHROMATOGRAPHIC ANALYSIS LIMITATION OF GC APPLICATION OF GC INTRODUCTION Gas chromatography is an instrumental /Flow Meters :- Rota meter & Soap bubble flow meter Stationary Phase The most common stationary phases in gas-chromatography columns are polysiloxanes , which contain various substituent groups to change the polarity of the phase. The nonpolar end of/


Ic_sampl_prep_26062003_e.ppt 1 IC Metrohm inline sample preparation Techniques for analytical success Metrohm Ion Chromatography Bart Cleeren, Metrohm.

no pre concentration Unique technique, that had significantly improves the field of application for IC ic_sampl_prep_26062003_e.ppt 14 IC Metrohm Ion Chromatography Dialysis: milk Inline sample preparation Metrosep Dual 1 – 70 1Chloride938 2Nitraten./ic_sampl_prep_26062003_e.ppt 23 IC P3 Metrohm Ion Chromatography Inline sample preparation # 1 Preconcentrator column inline EMobile phase: eluent P1High pressure pump Injection valve «A» CAnalytical column »MSM» Suppressor DConductivity detector P2Peristaltic /


Chapter 32 HIGH-PERRORMANCE LIQUID CHROMATOGRAPHY High-performance liquid chromatography (HPLC) is the most versatile and widely used type of elution chromatography.

screw- driven syringe type, (ii) a reciprocating pump, and (iii) a pneumatic or constant-pressure pump. Columns for High-Performance Liquid Chromatography Liquid-chromatographic columns are usually constructed from stainless steel tubing, although glass or Tygon tubing is sometimes employed for lower pressure application (<600 psi). Most columns range in length from 10 to 30 cm and have inside diameters of 4 to 10 mm/


Chromatography Russian scientist Tswett in 1906 used a glass columns packed with finely divided CaCO3 to separate plant pigments extracted by hexane. The.

future. 5- Monitor both column chromatography and organic chemical reactions. b- Quantities Chromatography: The development of modern instruments enable the use of chromatography to determine the amount of any component in a mixture as absolute amount or relative to another component HPLC/ GC/ HPTLC can be used for there applications. 2- Preparative application: This was the first and is the main application of chromatography. The technique was developed/


LIQUID CHROMATOGRAPHY CHAPTER 21 pg. 604 Adsorption Chromatography.

ligands. 2. eliminate the analyte ligand interaction. Change of pH or μ A: Extraordinary specificity Application: rapid isolation of biomolecules 4.Hydrophobic Interaction Chromatography Instruments 1.HPLC 2.FPLC HPLC FPLC 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/


Basics of Chromatography Jony Mallik. Introductory Principles Chromatography is a combination of two words; * Chromo – Meaning color * Graphy – representation.

min). Linear velocity: Distance passed by mobile phase per 1 min in the column (cm/min). Linear velocity: Distance passed by mobile phase per 1 min in the column (cm/min). Mobile Phase – gas or liquid that carries the mixture /linked dextrans (sephadex) Agarose (sepharose) Polyacrylamide Porous glass gel. APPLICATIONS Fractionation (purification of the desired protein using suitable gel) Molecular weight determination ION EXCHANGE Ion exchange chromatography is used to remove ions of one type from a mixture /


CHROMATOGRAPHY. CHROMATOGRAPHY This method is used for separation of mixtures of compounds between two phases:  Stationary ( solid or liquid located.

chromatography  Ion exchange chromatography  Partition chromatography  Gel (size exclusion) chromatography  Affinity chromatography  Capillary chromatography 2. Because of techniques of preparation:  Column chromatography  Planar chromatography ( thin layer, paper, gel) 3. Because of density of mobile phase:  GC, Gas Chromatography)  LC, Liquid Chromatography/ temperature have a significant effect on the output. Chromatography application for quantitative analysis – R f coefficient. Substance /


Over view What is ion chromatography? Why this techniqu is useful?

with system hardware as in all hightech areas Robustness - pH and solvent compatible separators allow a variety of applications - Analysis of complex matrices such as waste water, foods, body fluids, etc. History Aristol experiment(384-/operation – no extra programming Simplified hardware – no extra valves Flexible method development Single ion chromatography Used when –Exchange capacity of the separator column is low –Dilute eluents are employed –Ion suppression is unnecessary Examples of resins with low/


Chapter 33 Supercritical-Fluid Chromatography Supercritical-fluid chromatography (SFC), in which the mobile phase is a supercritical fluid, is a hybrid.

Chromatography versus Other Column Methods Several physical properties of supercritical fluids are intermediate between the properties of gases and liquids. As a consequence, this new type of chromatography combines some of the characteristics of both gas and liquid chromatography. Thus, like gas chromatography, supercritical-fluid chromatography is inherently faster than liquid chromatography because of the lower viscosity and higher diffusion rates in the mobile phase. Applications It is applicable/


Proteins separation and analysis using Fast Protein Liquid Chromatography Ayelet David, Ph.D Dept. of Clinical Pharmacology.

Chromatography (FPLC) Column chromatography used to separate or purify proteins from complex mixtures based on size, charge distribution, hydrophobicity of biorecognition (affinity chromatography). Typical columns used for protein purification, include: Gel Filtration Chromatography — separate proteins according to their size. Also termed as “size exclusion chromatography” (SEC). Ion exchange chromatography/> 1 Adsorption has occurred Some typical applications for gel filtration 1.Group separation: Desalting/


Chapter 33 High-Performance Liquid Chromatography.

is that no special equipment is required for suppression. However, it is a somewhat less sensitive method for determining anions than suppressor-column methods. 33 E Size-exclusion chromatography Size-exclusion, or gel chromatography, is a powerful technique that is particularly applicable to high-molecular-mass species. Fractionation is based on molecular size and to some extent molecular shape. The average residence time of/


Size exclusion Chromatography. Also called: Gel filtration chromatography (MP water): desalting Gel permeation chromatography (MP organic solvent) Molecules.

in solution (hydrodynamic volume) Species can be: single molecules polymer coils aggregates micelles etc… Applications Typical SEC applications: analysis of synthetic polymers and oligomers lipids proteins cellulose derivatives crude oil alkanes SEC methods can/ S, bonded silica. Conventional High Performance Conventional Cheap, Slow flow rate, Available in bulk For open column chromatography High Performance Expensive, Rapid flow rate, Available ready-to-use, For HPLC Separation of proteins Mobile Phases /


CHROMATOGRAPHY. Chromatography Chromatography basically involves the separation of mixtures due to differences in the distribution coefficient of sample.

인정받고 있다. Classification of Chromatography - Paper Chromatography - LC (Liquid Chromatography) – TLC, HPLC - LC (Liquid Chromatography) – TLC, HPLC - GC (Gas Chromatography) - GC (Gas Chromatography) Liquid Column Chromatography A sample mixture is /column length Decrease column diameter Decrease flow-rate Pack column uniformly Use uniform stationary phase (packing material) Decrease sample size Select proper stationary phase Select proper mobile phase Use proper pressure Use gradient elution LC Application/


HIGH PERFORMANCE LIQUID CHROMATOGRAPHY Idea of Chromatgraphyyy The mixture of substances is dissolved in a liquid or a gas and flowed through a tube.

Chromatography 1.Liquid Column Chromatography 2.Gas Liquid / Solid Chromatography 3.Thin-layer Chromatography Michael Tswett discovered liquid chromatography in 1906 and till late 1950,liquid chromatography lacked the attributes of gas chromatography and was being done on columns,papers and thin layer plates. 11 Modern Liquid Chromatography/ vacuum at the other end of the chromatographic column 2.Applying high pressure on the liquid mobile phase. Application of higher pressure on liquid mobile phase seems /


Liquid Chromatography: Still Striving for High Efficiency Drexel University Literature Seminar by Anna Caltabiano 17 May 2010.

much lower than in HPLC. Liquid Chromatography (HPLC) instrument Solvent delivery Pump Autosampler Column Detector Injector The Chromatographic Process – Theoretical Considerations Elution in Column Chromatography MobilephaseStationaryphase t0t0t0t0 t1t1t1t1 t2t2t2t2 t4t4t4t4 t3t3t3t3 /columns. However, monolithic columns have limited stability at higher pressures, temperatures and pH. In addition, the limited variety of monolithic columns does not meet the demands for a wide range of industrial applications/


Chromatography Dr.Tawfeq A. Al-Howiriny Associate Professor

there may be one dominant mechanism, the modes are not mutually exclusive. The exact mode of chromatography operating in a given application is determined principally by the nature of the packing, though it must be appreciated that, while/ elutropic series was by trapped was recorded by Trapp, who found that the eluting power of series of solvents for substances adsorbed in columns such as silica gel decreased in order Pure water > methanol > ethanol > propanol > acetone > ethyle acetate > diethyle ether >/


GC-MS Gas Chromatography-Mass Spectrometry

of stainless steel & form of a coil Disadvantage: more sample cannot loaded 3.SCOT columns (Support coated open tubular column Improved version of Golay / Capillary columns, have small sample capacity Made by depositing a micron size porous layer of supporting material / Thermal Conductivity Detector Electron Capture Detector Application of GC Analysis of Halogenated Pesticides 4 10 7 11 6 8 12 13 9 14 1 5 15 17 18 3 16 2 2ppb in Water Schematic of a Gas Chromatography-Mass Spectrometry (GC-MS) Instrument /


Chromatography for Protein purification 1

protein, but some (clusters of) hydrophobic groups occur at surface of protein. Surface hydrophobic side chains can interact with hydrophobic groups for example attached to a column. HYDROPHOBIC CHROMATOGRAPHY Temperature Increasing temperature --> stronger hydrophobic interactions Sample (application) Column having high concentration of a salt promotes binding (for example ammonium sulfate just below the concentration that starts to precipitate protein). Elution of bound proteins Negative/


Chromatography Learning objective: To be able to describe the method of chromatography and its applications.

method of chromatography and its applications Chromatography Chromatography is used to separate pure substances from a mixture of substances, such as a cell extract.Chromatography is used/chromatography Column chromatography uses a glass column filled with a cellulose slurry. Large samples can be pumped through the column and the separated fractions can be collected for further experiments, so this is preparative chromatography as opposed to analytical chromatography.Column chromatography uses a glass column/


Prepared by : Mona Abo Hasera

chromatography Based on stationary phase Column Planar Based on mobile phase Liquid Gas Kinds of Chromatography Liquid Column Chromatography gel filtration ,ion exchange, affinity, adsorption, reverse phase, metal binding (column) 2. Gas Liquid Chromatography (column) planar Chromatography (Thin-layer & paper ) LIQUID COLUMN CHROMATOGRAPHY A sample mixture is passed through a column/exercise spotting on a sheet of filter paper.) 6. After application of samples let the spots dry. Meanwhile measure with a /


Four basic types of column chromatography where mobile phase is a liquid Partition Chromatography Bonded-Phase Liquid-Liquid Adsorption Chromatography.

Four basic types of column chromatography where mobile phase is a liquid Partition Chromatography Bonded-Phase Liquid-Liquid Adsorption Chromatography Liquid-Solid Ion-Exchange Chromatography Exclusion (or Gel) Chromatography General Advantages of LC Sensitivity Quantitative Separation of nonvolatile and/or thermally fragile compounds Wide applicability Applications of LC Source: Skoog, Holler, and Nieman, Principles of Instrumental Analysis, 5 th edition, Saunders College Publishing. LC Separation /


Supercritical Fluid (SCF)

compounds Can separate compounds beyond the volatility range of GC Compared to LC: Reduced analysis time column chromatography Comparison of MW elution ranges for different types of column chromatography Source: Skoog, Holler, and Nieman, Principles of Instrumental Analysis, 5th edition, Saunders College Publishing. Advantages of supercritical fluid chromatography (SFC) Applications in quality control and sample Minimizes use of organic solvents Detection down to ppb or μg/


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