|
|
- Miles Warren
- 6 years ago
- Views:
Transcription
1 CHROMATOGRAPHY 1
2 Chromatography - a physical method of mixture separation in which the components to be separated are distributed between two phases, one of which is stationary (stationary phase) while the other (the mobile phase) moves in a definite direction. 2
3 CHROMATOGRAPHY Stationary phase ( solid or liquid located on neutral medium) Mobile phase ( liquid or gas) 3
4 Chromatographic separation involves sample placing the test mixture on the liquid or solid stationary phase and then passing the liquid or gaseous mobile Stationary phase Mobile phase detector phase through it or above it, that is elution of the mixture from a stationary phase. The mixture of components with different ratios of division are eluted (migrate) at different speed. These differences in the speed of migration lead to the separation of components in time and space. 4 4
5 Investigated compound is located in mobile phase(solvent) and migrates with it across stationary phase which is porous medium: adsorbent, ion exchanger or molecular sieve. 5
6 Illustration of Chromatography Stationary Phase Separation Mixture Components Mobile Phase Affinity to Stationary Phase Components Affinity to Mobile Phase Blue white Red Yellow 6
7 Mobile phase or carrier - solvent moving through the column Stationary phase or adsorbents - substance that stays fixed inside the column Eluent - fluid entering the column Eluate - fluid exiting the column (that is collected in flasks) Elution - the process of washing out a compound through a column using a suitable solvent Analyte - mixture which individual components have to be separated and analyzed 7
8 Classification of chromtographic methods 1. According to physico chemical forces: Adsorption chromatography Ion exchange chromatography Partition chromatography Gel (size exclusion) chromatography Affinity chromatography Capillary chromatography 2. According to parameters of the proces: High Pressure/Performance Liquid Chromatography (HPLC) Fast protein liquid chromatography (FPLC) 8
9 Classification of chromtographic methods 3. According to applied techniques: Column chromatography Planar chromatography (thin layer, paper) 4. According to mobile phase type: Gas Chromatography, GC Liquid Chromatography, LC Supercritical Fluid Chromatography, SFC 5. According to mobile and stationary phases type: Gas liquid chromatography (GLC) Liquid liquid chromatography (LLC) Gas solid chromatography (GSC) Liquid - solid chromatography (LSC) 9
10 Choice of the chromatographic method depends on: the amount of analysed mixture kind of compounds complexity of separation process 10
11 Types of chromatography Liquid Chromatography separates liquid samples with a liquid solvent (mobile phase) and a column composed of solid beads (stationary phase) Gas Chromatography separates vaporized samples with an inert carrier gas (mobile phase) and a column composed of a liquid or of solid beads (stationary phase) 11
12 Types of chromatography Adsorption Chromatography uses solid material as stationary phase (adsorbents). The separation of the mixture is caused by a different affinity of individual components (adsorbates) of a mixture to the surface of stationary phase. Planar chromatography: Paper Chromatography separates liquid samples with a liquid solvent (mobile phase) on a paper strip (stationary phase) Thin-Layer Chromatography separates liquid samples with a liquid solvent (mobile phase). Glass plate is covered with a thin layer of aluminium oxide or silica gel (stationary phase) 12
13 Adsorption chromatography If the stationary phase is solid, chromatography is called adsorption chromatography. Separation of the mixture is caused by different affinities of mixture components (adsorbates) to the surface of the stationary phase known as adsorbent. Adsorption of analysed sample particles, can be induced by: - physical forces (physical adsorption) - chemical influence (chemisorption) 13
14 The effectiveness of adsorption depends on: kind of adsorbent and its properties solvent type properties of adsorbed molecules 14
15 Classification of adsorbents according to: 1. Adsorption activity: Weak ( starch, saccharose, talc) Middle ( calcium carbonate, sodium carbonate) Strong ( aluminium oxide, activated silica acid) 2. Chemical properties: acidic ( SiO 2 ) basic ( CaO) neutral ( activated carbon) amphoteric ( Al 2 O 3 ) 15
16 Classification of adsorbents 3. Chemical nature: Organic ( carbon, starch, saccharose, polyamides ) Inorganic ( Al 2 O 3, MgO, natural and synthetic silicates ) Mixed (talc + saccharose, CaCO 3 + talc ) Specific (silica gel with specific pores ) 4. Polarity : Highly polar ( aluminium oxide, silica oxide) Weak polar (calcium carbonate, MgO) Non-polar ( activated carbon, talc, graphite) 16
17 Solvents used in chromatogrphy are arranged in an eluotropic series, depending on the ability of adsorption with respect to the substances dissolved in them and the ability of elution. They are used to develop the chromatogram and to elute the separate components. 1. Hexane 2. Carbon tetrachloride (CCl 4 ) 3. Benzene 4. Diethyl ether 5. Acetone 6. Chloroform 7. Ethyl acetate 8. Ethanol 9. Methanol 10. Water 17
18 Properties of adsorbed molecules Adsorption ability of adsorbed molecules depends on:: Size and spatial orientation Number and location of double bonds Number of polar and nonpolar substituents Number and kind of functional groups The degree of adsorption of the adsorbate increases with: Number of double bonds Number of functional groups Number of substituents of the same type 18
19 Thin Layer Chromatography TLC (partition chromatography) Separation of the mixture depends on the difference in partition coefficient of the mixture s components between two non miscible phases, from which one is a liquid coated on a medium (stationary phase), and a second one is a mobile phase (liquid,gas). Partition Coefficient is equal to the ratio of the concentration of the same substance in the stationary phase and the mobile phase. Nernst law of partition: K = C s / C m K partition coefficient at equilibrium state, depends only on the temperature and properties substance-forming solutions and does not depend on the amount of solute. C s concentration of particular compound (solute) dissolved in stationary phase C m -concentration of solute dissolved in mobile phase 19
20 Thin-Layer Chromatography (TLC) This is a fast method for the initial orientation in the amount and relative contributions of the components in the mixture of organic compounds. Advantages of TLC: a short time to develop the chromatogram simple methods of process control minimum consumption of materials ease to perform and low costs Application of TLC: separation of amino acids, peptides, saccharides, nucleotides
21 Chromatography application for qualititive analysis R f coefficient. Substance location on chromatogram is characterized by R f values. (ratio of fronts) = retention factor R f = distance from START line to the middle of substance spot - A distance from START line to the End line of solvent B R f - has characteristic value for particular substance in specific process conditions R f <1 Height reached by solvent Invastigated sample END START A B 21
22 Preparation of chromatographic plates Glass plates mm thick covered with silica gel Investigated solution are placed on the plate using micropipette Chromatography chamber must be : leak proof saturated by solvent vapors 22
23 Development of chromatogram Development occurs during migration of solvent from start to the end. Solvent migrates because of: capillary forces ( up technique) gravity forces ( down technique) Development of chromatogram After removing plate from the chamber and drying special reagent (detector) reacts with separated compounds of the mixture and give coloured spots (detector example: mixture of concentrated sulphuric and acetic acid, anisic aldehyde and ethanol). 23
24 GAS CHROMATOGRAPHY Fast and effective method of separation for volatile compounds mixtures. Separated compound is carried out by gas (inert in relation to the stationary phase) through the column filled with stationary phase Depending on the stationary phase gas chromatography can be divided into: Partition gas chromatography ( stationary phase liquid on stationary carrier) Adsorption gas chromatography ( stationary phase adsorbent) Capillary gas chromatography ( stationary phase liquid is directly on the column walls.) 24
25 Scheme of gas chromatograph Sample s injection Chromatogram = result Carrier gas as mobile phase Oven Column Column with stationary phase (liquid on inert base or solid - adsorbent). Typical stationary phases are large molecular weight polysiloxane, polyethylene glycol, or polyester polymers of 0.1 to 2.5 micrometer film thickness. Columns are, available in many stationary phases sizes. A typical capillary column is 15 to 60 meters in length and 0.25 to 0.32 mm inner diameter. 25
26 Obtaining chromatogram in gas chromatography
27 GAS CHROMATOGRAPHY cont. Carrier gas must be chemically inert ( hydrogen, helium, nitrogen, argon, carbon dioxide) and is introduced to the column. Investigated sample or mixture is injected to the column with gas carrier. Stream of gas is carrying samples to be separated. Compounds are divided according to their retention factor between gas and stationary phase. Separated substances are measured and registered at the outlet of the column using detector system. Operating temperature is 0 o C 400 o C. Stationary phase micro-layer of organic liquids on a inert solid. 27
28 Detectors in gas chromatography Quantitative measurement of mixture components can be done by using different chemical and physical properties of the compounds. Therefore, there are the following types of detectors: 1. Thermal conductivity detector katharometer (TCD). 2. Flame-ionization detector - (FID). 3. Discharge ionization detector (DID). 4. Electron capture detector (ECD). Detector signals are registered by sensitive writing system. As a result we are obtaining CHROMATOGRAM. 28
29 Chromatogram is a visible record showing the result of separation of the components of a mixture by chromatography Czas Peak area is proportional to the amount of single analyzed sample. 29
30
31 HPLC Agilent Company 31
32 GAS CHROMATOGRAPHY Separation ability of column is increasing with length of the column, type and amount of stationary phase, column temperature, speed and pressure of carrier gas. Gas chromatography is applied in qualitative and quantitative chemistry (time retention and area or hight of the peak). 32
33 Gas chromatograph 33
34 Chromatography column in gas chromatograph 34
35 Gel chromatography Used for separation of big molecules such as protein, polymers, nucleic acids, sugars, glycols, silicones etc. Column is filled with gel particles: polyacrylamide, polyethylene oxide, dextrin. As a mobile phase is used liquid. Separation is based on sieve and capillary mechanisms*. Large particles are moving fast and small particles are moving slowly. Method also used for separation of DNA molecules. 35
36 HPLC High Pressure/Performance Liquid Chromatography Mobile phase pressurized liquid solvent (20MPa-40MPa) Stationary phase solid adsorbent material Analytical version narrow columns (4-8 mm); length 5 25 cm Preparation version - wide columns ( mm); length cm Benefits: fast, automated and precise compound separation; easy appliacation of concentrated gradients; quantitative determination of the compounds Detector: spectroscopy (UV-vis), electrochemical, spectrofluorometric Drawback: separation only of small quantities HPLC columns 36
37 Column chromatography some time ago ;-) Chromatographic columns 37
38 It is used for the determination of: - Biologically active compounds such as amino acids, proteins, polysaccharides, vitamins, steroids and nucleic acids - Pharmaceutical Preparations - Plant protection products - pesticides - Polycyclic hydrocarbons -benzopyrene - Complexes - Rare earth elements 38
39 ADPR UA Urd Ino mau 10 VWD1 A, Wavelength=254 nm (US\16PUR051.D) GTP GDP NAD GMP 8 ADP ATP NADP Guo 6 IMP Hyp AMP 4 2 Xan min
40 THE END
Chromatography. writing in color
Chromatography writing in color Outlines of Lecture Chromatographic analysis» Principles and theory.» Definition.» Mechanism.» Types of chromatography.» Uses of Chromatography. In 1906 Mikhail Tswett used
More informationPRINCIPLES AND APPLICATION OF CHROMATOGRAPHY. Dr. P. Jayachandra Reddy Mpharm PhD Principal & professor KTPC
PRINCIPLES AND APPLICATION OF CHROMATOGRAPHY Dr. P. Jayachandra Reddy Mpharm PhD Principal & professor KTPC CHROMATOGRAPHY Laboratory technique for the Separation of mixtures Chroma -"color" and graphein
More informationCHROMATOGRAPHY. The term "chromatography" is derived from the original use of this method for separating yellow and green plant pigments.
CHROMATOGRAPHY The term "chromatography" is derived from the original use of this method for separating yellow and green plant pigments. THEORY OF CHROMATOGRAPHY: Separation of two sample components in
More informationCourse goals: Course goals: Lecture 1 A brief introduction to chromatography. AM Quality parameters and optimization in Chromatography
Emqal module: M0925 - Quality parameters and optimization in is a separation technique used for quantification of mixtures of analytes Svein.mjos@kj.uib.no Exercises and lectures can be found at www.chrombox.org/emq
More informationInstrumental Chemical Analysis
L2 Page1 Instrumental Chemical Analysis Chromatography (General aspects of chromatography) Dr. Ahmad Najjar Philadelphia University Faculty of Pharmacy Department of Pharmaceutical Sciences 2 nd semester,
More informationGas Chromatography. Presented By Mr. Venkateswarlu Mpharm KTPC
Gas Chromatography Gas Chromatography Presented By Mr. Venkateswarlu Mpharm KTPC What is Gas Chromatography? It is also known as Gas-Liquid Chromatography (GLC) GAS CHROMATOGRAPHY Separation of gaseous
More informationChromatography. Gas Chromatography
Chromatography Chromatography is essentially the separation of a mixture into its component parts for qualitative and quantitative analysis. The basis of separation is the partitioning of the analyte mixture
More informationWhat is Chromatography?
What is Chromatography? Chromatography is a physico-chemical process that belongs to fractionation methods same as distillation, crystallization or fractionated extraction. It is believed that the separation
More informationOpen Column Chromatography, GC, TLC, and HPLC
Open Column Chromatography, GC, TLC, and HPLC Murphy, B. (2017). Introduction to Chromatography: Lecture 1. Lecture presented at PHAR 423 Lecture in UIC College of Pharmacy, Chicago. USES OF CHROMATOGRAPHY
More informationChromatography. Chromatography is a combination of two words; * Chromo Meaning color * Graphy representation of something on paper (writing)
Chromatography Chromatography is a combination of two words; * Chromo Meaning color * Graphy representation of something on paper (writing) Invention of Chromatography Mikhail Tswett invented chromatography
More informationCHEMISTRY Unit 3, Area of Study 1: Chemical Analysis
Watch this lesson online: https://edrolo.com.au/vce/subjects/chemistry/vce-chemistry/aos-1-chemical-analysis/chromatography-hplc-glc/column-chromatography/#watch CHEMISTRY Unit 3, Area of Study 1: Chemical
More informationDEFINITION CHROMATOGRAPHY
Chromatography DEFINITION CHROMATOGRAPHY The separation of a mixture by distribution of its components between a mobile and stationary phase over time mobile phase = solvent stationary phase = column packing
More informationIntroduction to Chromatography
Introduction to Chromatography Dr. Sana Mustafa Assistant Professor Department of Chemistry, Federal Urdu University of Arts, Science & Technology, Karachi. What is Chromatography? Derived from the Greek
More informationChapter 1. Chromatography. Abdul Muttaleb Jaber
Chapter 1 Chromatography Abdul Muttaleb Jaber What is Chromatography? Chromatography is a physico-chemical process that belongs to fractionation methods same as distillation, crystallization or fractionated
More informationChromatographic Separation
What is? is the ability to separate molecules using partitioning characteristics of molecule to remain in a stationary phase versus a mobile phase. Once a molecule is separated from the mixture, it can
More informationInstrumental Analysis II Course Code: CH3109. Chromatographic &Thermal Methods of Analysis Part 1: General Introduction. Prof. Tarek A.
Instrumental Analysis II Course Code: CH3109 Chromatographic &Thermal Methods of Analysis Part 1: General Introduction Prof. Tarek A. Fayed What is chemical analysis? Qualitative analysis (1) Chemical
More informationLab.2. Thin layer chromatography
Key words: Separation techniques, compounds and their physicochemical properties (molecular volume/size, polarity, molecular interactions), mobile phase, stationary phase, liquid chromatography, thin layer
More informationChapter content. Reference
Chapter 7 HPLC Instrumental Analysis Rezaul Karim Environmental Science and Technology Jessore University of Science and Technology Chapter content Liquid Chromatography (LC); Scope; Principles Instrumentation;
More informationCH 2252 Instrumental Methods of Analysis Unit V Gas Chromatography. M. Subramanian
CH 2252 Instrumental Methods of Analysis Unit V Gas Chromatography M. Subramanian Assistant Professor Department of Chemical Engineering Sri Sivasubramaniya Nadar College of Engineering Kalavakkam 603
More informationExperiment 1: Thin Layer Chromatography
Experiment 1: Thin Layer Chromatography Part A: understanding R f values Part B: R f values & solvent polarity Part C: R f values & compound functionality Part D: identification of commercial food dye
More informationCHAPTER 6 GAS CHROMATOGRAPHY
CHAPTER 6 GAS CHROMATOGRAPHY Expected Outcomes Explain the principles of gas chromatography Able to state the function of each components of GC instrumentation Able to state the applications of GC 6.1
More informationIntroduction to Gas Chromatography
Introduction to Gas Chromatography 31-1 Objectives To know what is chromatography To understand the mechanism of compound separation To know the basic of gas chromatography system 31-2 Chromatography Definition
More informationHigh Performance Liquid Chromatography
High Performance Liquid Chromatography What is HPLC? It is a separation technique that involves: Injection of small volume of liquid sample Into a tube packed with a tiny particles (stationary phase).
More informationHigh Pressure/Performance Liquid Chromatography (HPLC)
High Pressure/Performance Liquid Chromatography (HPLC) High Performance Liquid Chromatography (HPLC) is a form of column chromatography that pumps a sample mixture or analyte in a solvent (known as the
More informationGas Chromatography. Vaporization of sample Gas-solid Physical absorption Gas-liquid Liquid immobilized on inert solid
Gas Chromatography Vaporization of sample Gas-solid Physical absorption Gas-liquid Liquid immobilized on inert solid Principles Instrumentation Applications 18-1 Retention Volumes Volumes rather than times
More informationChromatography and Functional Group Analysis
Chromatography Chromatography separates individual substances from a mixture. - to find out how many components there are - to match the components with known reference materials - to use additional analytical
More informationChromatographic Methods of Analysis Section: 5 Gas Chromatography (GC) Prof. Tarek A. Fayed
Chromatographic Methods of Analysis Section: 5 Gas Chromatography (GC) Prof. Tarek A. Fayed Gas Chromatography (GC) In gas chromatography, the sample is vaporized and injected onto the head of a chromatographic
More informationChromatography- Separation of mixtures CHEM 212. What is solvent extraction and what is it commonly used for?
Chromatography- Separation of mixtures CHEM 212 What is solvent extraction and what is it commonly used for? How does solvent extraction work? Write the partitioning coefficient for the following reaction:
More informationStationary phase: Non-moving phase that provides support for mixture to move.
Chromatography For S. Y. B. Sc. Sem-IV By: Dr Vipul B. Kataria The technique was invented by Russian botanist Mikhail tswett in 1906. The word Chromatography is derived from latin language and in this
More informationPrinciples of Gas- Chromatography (GC)
Principles of Gas- Chromatography (GC) Mohammed N. Sabir January 2017 10-Jan-17 1 GC is a chromatographic technique utilizes gas as the mobile phase which is usually an inert gas (Hydrogen, Helium, Nitrogen
More informationChapter 27: Gas Chromatography
Chapter 27: Gas Chromatography Gas Chromatography Mobile phase (carrier gas): gas (He, N 2, H 2 ) - do not interact with analytes - only transport the analyte through the column Analyte: volatile liquid
More informationLuminescence transitions. Fluorescence spectroscopy
Luminescence transitions Fluorescence spectroscopy Advantages: High sensitivity (single molecule detection!) Measuring increment in signal against a dark (zero) background Emission is proportional to excitation
More informationCHEM 429 / 529 Chemical Separation Techniques
CHEM 429 / 529 Chemical Separation Techniques Robert E. Synovec, Professor Department of Chemistry University of Washington Lecture 1 Course Introduction Goal Chromatography and Related Techniques Obtain
More informationChromatography & instrumentation in Organic Chemistry
Chromatography & instrumentation in Organic Chemistry What is Chromatography? Chromatography is a technique for separating mixtures into their components in order to analyze, identify, purify, and/or quantify
More informationINSTITUTE OF MEDICAL BIOCHEMISTRY FIRST FACULTY OF MEDICINE, CHARLES UNIVERSITY IN PRAGUE. Chromatography. in biochemistry
INSTITUTE OF MEDICAL BIOCHEMISTRY FIRST FACULTY OF MEDICINE, CHARLES UNIVERSITY IN PRAGUE Chromatography in biochemistry Prof. RNDr. Věra Pacáková, CSc. Faculty of Natural Sciences, Charles University
More informationCHROMATOGRAPHY AND MASS SPECTROMETER
22 CHROMATOGRAPHY AND MASS SPECTROMETER 22.1 INTRODUCTION We know that the biochemistry or biological chemistry deals with the study of molecules present in organisms. These molecules are called as biomolecules
More informationAbstract: An minimalist overview of chromatography for the person who would conduct chromatographic experiments, but not design experiments.
Chromatography Primer Abstract: An minimalist overview of chromatography for the person who would conduct chromatographic experiments, but not design experiments. At its heart, chromatography is a technique
More informationChromatography. What is Chromatography?
Chromatography What is Chromatography? Chromatography is a technique for separating mixtures into their components in order to analyze, identify, purify, and/or quantify the mixture or components. Mixture
More informationChromatography Outline
Chem 2001 Summer 2004 Outline What is? The Chromatogram Optimization of Column Performance Why Do Bands Spread? Gas High-Performance Liquid Ion-Exchange 2 What is? In chromatography, separation is achieved
More informationPhysical Separations and Chromatography
Lab #5A & B: Physical Separations and Chromatography Individual Objectives: At the end of these experiments you should be able to: Ø Distinguish between Rf and tr; chromatograph and chromatogram; adsorption
More informationGC Instruments. GC Instruments - Sample Introduction
GC Instruments 1 Fairly simple instrumentation Maintaining constant average pressure is important! Pressure controls flow rate T influences retention (k ) Flow rate monitoring Changing flow rate changes
More informationChromatographie Methods
Chromatographie Methods Fifth Edition A. BRAITHWAITE Department of Physical Sciences Nottingham Trent University and F. J. SMITH Department of Chemistry and Chemical Engineering University of Paisley BLACKIE
More informationGas Chromatography (GC)! Environmental Organic Chemistry CEE-PUBH Analysis Topic 5
Gas Chromatography (GC)! Environmental Organic Chemistry CEE-PUBH 5730-6730 Analysis Topic 5 Chromatography! Group of separation techniques based on partitioning (mobile phase/stationary phase). Two immiscible
More informationGas Chromatography (GC)
Gas Chromatography (GC) Ahmad Aqel Ifseisi Assistant Professor of Analytical Chemistry College of Science, Department of Chemistry King Saud University P.O. Box 2455 Riyadh 11541 Saudi Arabia Office: AA53
More informationGAS CHROMATOGRAPHY. Mobile phase is a gas! Stationary phase could be anything but a gas
GAS CHROMATOGRAPHY Mobile phase is a gas! Stationary phase could be anything but a gas Gas Chromatography (GC) GC is currently one of the most popular methods for separating and analyzing compounds. This
More informationChapter 23 Introduction to Analytical Separations
Chapter 23 Introduction to Analytical Separations Homework Due Monday April 24 Problems 23-1, 23-2, 23-7, 23-15, 23-27, 23-29, 23-32 Analytical Separations: Universal approach to analyzing complex mixtures
More informationChromatography. Mrs. D. MEENA MPharm PA & QA KTPC
Chromatography Mrs. D. MEENA MPharm PA & QA KTPC INTRODUCTION ANALYTICAL TECHNIQUES Analytical chemistry involves separating, identifying and determining the relative amount of the components in a sample
More informationIntroduction to Chromatographic Separations
Introduction to Chromatographic Separations Analysis of complex samples usually involves previous separation prior to compound determination. Two main separation methods instrumentation are available:
More informationCHIRAL SEPARATION USING THIN LAYER CHROMATOGRAPHY
CHIRAL SEPARATION USING THIN LAYER CHROMATOGRAPHY Chiral Chromatography Chiral - adjective: not superimposable on its mirror image: used to describe a molecule whose arrangement of atoms is such that it
More informationCfE Higher Chemistry. Unit 3: Chemistry in Society. Chemical Analysis as part of quality control
CfE Higher Chemistry Unit 3: Chemistry in Society Chemical Analysis as part of quality control 06/12/2017 Composition and purity 06/12/2017 Learning Outcomes : I can explain the basic principle of how
More informationPrinciples of Instrumental Analysis
Principles of Instrumental Analysis Chapter 27 Gas Chromatography Gas Chromatography (GC): vaporized analytes (solutes) are partitioned between a mobile gaseous phase and a liquid or a solid stationary
More informationDownstream Processing Prof. Mukesh Doble Department Of Biotechnology Indian Institute of Technology, Madras. Lecture - 33 HPLC
Downstream Processing Prof. Mukesh Doble Department Of Biotechnology Indian Institute of Technology, Madras Lecture - 33 HPLC Today, we are going to talk about the HPLC. HPLC is an analytical tool, which
More informationChromatography Lab # 4
Chromatography Lab # 4 Chromatography is a method for separating mixtures based on differences in the speed at which they migrate over or through a stationary phase which means that a complex mixture will
More informationBiochemistry. Biochemical Techniques. 12 Gas Liquid Chromatography
Description of Module Subject Name Paper Name 12 Module Name/Title 12 Gas - liquid Chromatography 1. Objectives 1.1 To understand principle of Gas Liquid Chromatography 1.2 To explain the different components
More informationCh.28 HPLC. Basic types of Liquid Chromatography Partition (LLC) Adsorption (LSC) Ion Exchange (IC) Size Exclusion (SEC or Gel Chromatography)
Ch.28 HPLC 28.1 Basic types of Liquid Chromatography Partition (LLC) Adsorption (LSC) Ion Exchange (IC) Size Exclusion (SEC or Gel Chromatography) High Performance (Pressure) LC Glass column st.steel (high
More informationChapter 27: Gas Chromatography. Principles Instrumentation Detectors Columns and Stationary Phases Applications
Chapter 27: Gas Chromatography Principles Instrumentation Detectors Columns and Stationary Phases Applications GC-MS Schematic Interface less critical for capillary columns Several types of Mass Specs
More informationPartitioning. Separation is based on the analyte s relative solubility between two liquid phases or a liquid and solid.
Chromatography Various techniques for the separation of complex mixtures that rely on the differential affinities of substances for a gas or liquid mobile medium and for a stationary adsorbing medium through
More informationExperiment Nine Thin Layer Chromatography
Name: Lab Section: 09 Thin Layer Chromatography Experiment Nine Thin Layer Chromatography Introduction Objective Separation of compounds from a mixture is an incredibly important aspect of experimental
More information2401 Gas (liquid) Chromatography
2401 Gas (liquid) Chromatography Chromatography Scheme Gas chromatography - specifically gas-liquid chromatography - involves a sample being vaporized and injected onto the head of the chromatographic
More informationHPLC Background Chem 250 F 2008 Page 1 of 24
HPLC Background Chem 250 F 2008 Page 1 of 24 Outline: General and descriptive aspects of chromatographic retention and separation: phenomenological k, efficiency, selectivity. Quantitative description
More informationChapter 26. An Introduction to Chromatographic Separations. Chromatography
Chapter 26 An Introduction to Chromatographic Separations Chromatography 1 Chromatography-Model as Extraction Chromatography-Model as Extraction 2 Chromatography Planar Chromatography-Types paper chromatography
More informationChromatography: Thin-Layer Chromatography (TLC) & Column Chromatography
Chromatography: Thin-Layer Chromatography (TLC) & Column Chromatography Part 1, p. 184: Separation of spinach pigments by TLC. (4 th Ed. P. 180) Part 2, p. 192: Separation of Fluorene and Fluorenone by
More informationVolatile organic compounds (VOCs):
Volatile organic compounds (VOCs): Organic chemicals with a high vapour pressure at room temperature. High vapour pressure results from a low boiling point. The World Health Organization (WHO) defined
More informationLiquid Chromatography
Liquid Chromatography 1. Introduction and Column Packing Material 2. Retention Mechanisms in Liquid Chromatography 3. Method Development 4. Column Preparation 5. General Instrumental aspects 6. Detectors
More informationAnalytical Chemistry
Analytical Chemistry Chromatographic Separations KAM021 2016 Dr. A. Jesorka, 6112, aldo@chalmers.se Introduction to Chromatographic Separations Theory of Separations -Chromatography Terms Summary: Chromatography
More informationDisadvantage: Destructive Technique once analyzed by GC, the sample is lost
Gas Chromatography Like other methods of chromatography, a partitioning of molecules must occur between the stationary phase and the mobile phases in order to achieve separation. This is the same equilibrium
More informationHarris: Quantitative Chemical Analysis, Eight Edition CHAPTER 23: GAS CHROMATOGRAPHY
Harris: Quantitative Chemical Analysis, Eight Edition CHAPTER 23: GAS CHROMATOGRAPHY Chapter 23. Gas Chromatography What did they eat in the year 1,000? GC of Cholesterol and other lipids extracted from
More information10/27/10. Chapter 27. Injector typically 50 C hotter than oven
Sample and solvent are vaporized onto the head of a column Vaporized solvent and solute are carried through the column by an inert gas (mobile phase) The mobile phase does not interact with compounds of
More informationSkoog/Holler/Crouch Chapter 26 Principles of Instrumental Analysis, 6th ed. CHAPTER 26
Skoog/Holler/Crouch Chapter 26 Principles of Instrumental Analysis, 6th ed. Instructor s Manual CHAPTE 26 26-1. (a) Elution is a process in which species are washed through a chromatographic column by
More informationChapter 31 Gas Chromatography. Carrier Gas System
Chapter 31 Gas Chromatography GAS-LIQUID CHROMATOGRAPHY In gas chromatography, the components of a vaporized sample are fractionated as a consequence of being partitioned between a mobile gaseous phase
More informationAnalytical Technologies in Biotechnology Prof. Dr. Ashwani K. Sharma Department of Biotechnology Indian Institute of Technology, Roorkee
Analytical Technologies in Biotechnology Prof. Dr. Ashwani K. Sharma Department of Biotechnology Indian Institute of Technology, Roorkee Module - 3 Chromatographic Method Lecture - 1 Introduction and Basic
More informationCh24. Gas Chromatography (GC)
Ch24. Gas Chromatography (GC) 24.1 What did they eat in the year 1000? From 13 C content of cholesterol in ancient bone 13 C : 1.1%, 12 C: 98.9% 13 C/ 12 C ratio types of plants Bones of 50 people in Barton-on-Humber
More informationWhat type of samples are common? Time spent on different operations during LC analyses. Number of samples? Aims. Sources of error. Sample preparation
What type of samples are common? Sample preparation 1 2 Number of samples? Time spent on different operations during LC analyses 3 4 Sources of error Aims Sample has to be representative Sample has to
More informationLEARNING OBJECTIVES CHEM 212: SEPARATION SCIENCE CHROMATOGRAPHY UNIT. Thomas Wenzel, Bates College. In-class Problem Set Extraction.
LEARNING OBJECTIVES CHEM 212: SEPARATION SCIENCE CHROMATOGRAPHY UNIT Thomas Wenzel, Bates College In-class Problem Set Extraction Problem #1 1. Devise a scheme to be able to isolate organic acids, bases
More informationCHROMATOGRAPHY (I): BASIS OF ELEMENTAL CHROMATOGRAPHY
Theme 06. CHROMATOGRAPHY (I): CHROMATOGRAPHY ELEMENTARY BASES 1599-ENVIRONMENTAL ANALYTIC METHODS Grade in Environmental Sciences Course 2013-14 Second Semester Professors: Miguel A Sogorb (msogorb@umh.es)
More informationPAPER AND THIN LAYER CHROMATOGRAPHY (TLC)
PAPER AND THIN LAYER CHROMATOGRAPHY (TLC) Objectives Understand the principle of Paper and Thin Layer Chromatography (TLC). Diagnose two samples of urine for Phenylketonuria and Cystinuria, using paper
More informationHPLC Workshop 16 June 2009 What does this do? Chromatography Theory Review Several chromatographic techniques Even though each method utilizes different techniques to separate compounds, the principles
More informationPackings for HPLC. Packings for HPLC
Summary of packings for HPLC In analytical HPLC, packings with particle sizes of 3 to 10 µm are preferred. For preparative separation tasks, also particles with diameters larger than 10 µm are applied.
More informationPrelab Reading Assignment: Laboratory Techniques in Organic Chemistry, 4 th Ed. Chapter 19
CHEM 213 Technique Experiments Experiment 5: Column Chromatography Number of labs - one Reactions performed None Chemicals used: Fluorene-fluorenone mixture, hexanes, methylene chloride, silica gel Supplies
More informationLAB #6 Chromatography Techniques
LAB #6 Chromatography Techniques Objectives: To learn how to story board a procedure Explain how a chromatograph of pigments is formed from both paper and thin layer chromatography. Isolate and identify
More informationGas Chromatography. Introduction
Gas Chromatography Introduction 1.) Gas Chromatography Mobile phase (carrier gas) is a gas - Usually N 2, He, Ar and maybe H 2 - Mobile phase in liquid chromatography is a liquid Requires analyte to be
More informationSeparation Techniques and Extraction methods of Phytochemicals
Separation Techniques and Extraction methods of Phytochemicals Introduction herbal medicines and the products derived from them are traditionally used for their benefits in prevention and treatment of
More informationChapter 11 Conventional Gas Chromatography
Chapter 11 Conventional Gas Chromatography Gas Chromatography GC is the first instrumental chromatographic method developed commercially It is relatively easy to introduce a stable flow and pressure for
More information7 INSTRUMENTAL CHROMATOGRAPHY
7 INSTRUMENTAL CHROMATOGRAPHY 7.1 Introduction There are two forms of chromatography, very widely used in analytical laboratories, which rely on electronic control of the process and detection of the species.
More informationChromatographic Methods of Analysis Section 2: Planar Chromatography. Prof. Tarek A. Fayed
Chromatographic Methods of Analysis Section 2: Planar Chromatography Prof. Tarek A. Fayed Planar chromatography includes two types: 1- Thin Layer Chromatography (TLC). 2- Paper Chromatography (PC). Thin
More informationChemistry Instrumental Analysis Lecture 28. Chem 4631
Chemistry 4631 Instrumental Analysis Lecture 28 Two types in general use: -packed (stationary phase) -open tubular or capillary determine selectivity and efficiency of the sample. Column Materials Column
More informationLab 3 Guide: Gas Chromatography (GC) (Sept 8-14)
Lab 3 Guide: Gas Chromatography (GC) (Sept 8-14) How GC works The Basic Idea Gas chromatography (GC) is mainly used for the qualitative analysis of samples: it answers the question What chemicals are present
More informationLIQUID CHROMATOGRAPHY
LIQUID CHROMATOGRAPHY RECENT TECHNIQUES HPLC High Performance Liquid Chromatography RRLC Rapid Resolution Liquid Chromatography UPLC Ultra Performance Liquid Chromatography UHPLC Ultra High Pressure Liquid
More informationChromatographic Analysis
Chromatographic Analysis Distribution of Analytes between Phases An analyte is in equilibrium between the two phases [S 1 ] [S 2 ] (in phase 1) (in phase 2) AS [S2 ] K 2 A S [S1 ] 1 AS, A 1 S Activity
More informationnot to be republished NCERT THE technique of chromatography is vastly used for the separation, Chromatography UNIT-5 EXPERIMENT 5.
UNIT-5 Chromatography THE technique of chromatography is vastly used for the separation, purification and identification of compounds. According to IUPAC, chromatography is a physical method of separation
More informationSample Preparation TLC Plates
TLC PLATES TLC Plates Economical separation method High sample throughput Pilot procedure for HPLC and flash chromatography Versatile range of ready-to-use layers Thin layer chromatography (TLC) is a simple,
More informationChromatography and its applications
Chromatography and its applications Reference Introduction to chromatography theory and practice Instrumental methods of chemical analysis by H.Kaur INTRODUCTION There are many methods which have been
More informationCourse CHEM Chromatography
Course CHEM 340 - Chromatography - Chromatographic Methods o Gas Chromatography (GC) o High performance Liquid Chromatography (HPLC) Terms Stationary phase A fixed place either in a column or on a planer
More informationAn Advanced Base Deactivated Capillary Column for analysis of Volatile amines Ammonia and Alcohols.
An Advanced Base Deactivated Capillary Column for analysis of Volatile amines Ammonia and Alcohols. Jaap de Zeeuw, Ron Stricek and Gary Stidsen Restek Corp Bellefonte, USA To analyze basic compounds at
More informationGC Instruments. GC Instruments - Columns
GC Instruments 1 Fairly simple instrumentation Maintaining constant average pressure is important! Pressure controls flow rate T influences retention (k ) Flow rate monitoring Changing flow rate changes
More informationChemistry Instrumental Analysis Lecture 28. Chem 4631
Chemistry 4631 Instrumental Analysis Lecture 28 High Performance Liquid Chromatography () Instrumentation Normal Phase Chromatography Normal Phase - a polar stationary phase with a less polar mobile phase.
More informationGas chromatography. Advantages of GC. Disadvantages of GC
Advantages of GC Gas chromatography Fast analysis, typically minutes Effi cient, providing high resolution Sensitive, easily detecting ppm and often ppb Nondestructive, making possible on - line coupling;
More informationMethods of pollution control and waste management - laboratory. Adsorptive removal of volatile organic compounds from gases streams
Methods of pollution control and waste management - laboratory Adsorptive removal of volatile organic compounds from gases streams Manual for experiment 17 dr Hanna Wilczura-Wachnik and dr inż. Jadwiga
More information31. Applications of the chromatographic methods in drug analysis
31. Applications of the chromatographic methods in drug analysis 31. 1. Depending on the processes that underlie the chromatographic separation, the following methods exist: A. adsorption chromatography
More informationFall 2012 Due In Class Friday, Oct. 19. Complete the following on separate paper. Show your work and clearly identify your answers.
CHEM 322 Name Fall 2012 Due In Class Friday, Oct. 19 Complete the following on separate paper. Show your work and clearly identify your answers. General Separations 1. Describe the relative contributions
More information