High Temperature Ultra Fast Liquid Chromatography. Peter W. Carr, Jon Thompson, Dwight Stoll, and Adam Schallinger
|
|
- Karen Patterson
- 5 years ago
- Views:
Transcription
1 High Temperature Ultra Fast Liquid Chromatography Peter W. Carr, Jon Thompson, Dwight Stoll, and Adam Schallinger
2 Conclusions 1. To do fast LC, use a WEAK eluent and a HOT column.. Use a highly retentive column so that you can work at lowest possible viscosity!
3 A. ma U Fast Separation of Antihistamines at 80 o C V WD1 A, Wavelength=54 nm (P10501\TEST0146.D) Temperature: 1 o C Flow rate: 1.35 ml/min. Pressure drop: 195 bar Resolution (,3):.1 Analysis time: 1.5 minutes min min ma U B V WD1 A, Wavelength=54 nm (P10501\TEST0151.D) 1 3 1=Doxylamine, =Methapyrilene, 3=Chlorpheniramine, 4=Triprolidine, 5=Meclizine4=Triprolidine, 5=Meclizine Temperature: 80 o C Flow rate: 3.00 ml/min. Pressure drop: 195 bar Resolution (,3):.1 Analysis time:.5 minutes min min LC Conditions: (A) Mobile Phase, 9/71 ACN/50mM Tetramethylammonium hydroxide; Injection volume, 0.5 ul; 54 nm detection; 100 x 4.6 ZirChrom-PBD (B) same as A, except Mobile Phase, 6.5/73.5 ACN/50mM Tetramethylammonium hydroxide, ph 1.
4 Faster Separation at 140 o C 100 mau Temperature: 1 o C Flow rate: 1.35 ml/min. Pressure drop: 195 bar Resolution (,3):.1 Analysis time: 1.5 min Time (min.) mau Temperature: 140 o C Flow rate: 4.0 ml/min. Pressure drop: 194 bar Resolution (,3):.1 Analysis time: 0.85 minutes Time (min.) Mobile Phase, 9/71 ACN/50mM Tetramethylammonium hydroxide, ph 1.; Injection volume, 0.5 µl; 54 nm detection; Solutes: 1=Doxylamine, =Methapyrilene, 3=Chlorpheniramine, 4=Triprolidine, 5=Meclizine; Column, 100 mm x 4.6 mm i.d. ZirChrom -PBD
5 Effect of Temperature on Theoretical Analysis Time at Constant Pressure, Retention, and Plate Count* 1.0 t Analysis (T)/ t Analysis ( 5 o C) T ( o C) High-Performance Liquid Chromatography at Elevated Temperatures: Examination of Condition for the Rapid Separation of Large Molecules, R. D. Antia and Cs. Horvath, J. Chromatogr., 435, 1-15 (1988).
6 Relative Viscosity vs. Temperature η/η (5 o C) Water 0 % ACN/Water Methanol Acetonitrile Temperature ( o C)
7 How to Prevent Boiling Boiling Point of THF (Centigrade) Boiling Point Pressure (psi)
8 Requirements for High Temperature LC Stationary Phase Stability Thermal Mismatch Broadening On-Column Analyte Instability
9 Peak Shapes Observed for Various Mobile-Phase Feed Temperatures* σ obs = σ + σ σ column extra columnn + thermal mismatch LC conditions: Column at 30 o C; 6. mm IDx8cm; 3µ Zorbax ODS; at 5 ml/min; 50/50 (v/v) ACN,H O; nitrobenzene *H. Poppe and J.C. Kraak, J. Chromatogr., 8, (1983).
10 Comparison of the Effect of Incomplete Thermal Equilibration on Column Performance mm ID mm ID6 mm ID cm/min cm/min Absorbance (mau) 40 0 Absorbance (mau) Time (min) Time (min) LC conditions:.1 x 5 cm, C-18 INERT, 55 % ACN, 5 cm preheater, 60 o C 4.6 x 5 cm, C-18 INERT, 60% ACN, 5 cm preheater, 60 o C. Peaks: 1. toluene,. ethylbenzene, 3. propylbenzene,4. butylbenzene 0
11 Theoretical Effect of Temperature on Column Dynamics o C 75 o C H *10-5 (cm) o C 175 o C u*100 (cm/s) Conditions: The particle diameter is 3 µm and the reduced linear velocity does not change with temperature ((D m,5 o C = 6*10-7 cm /s). The linear velocity (u) is increased and the reduced plate height is calculated from a modified Knox equation (A = 1.5, B = 0.8, C = 0.3, D = 0.04) at each velocity and temperature. Fast desorption kinetics are assumed (E a = 0 kj/mol, k o = 1*10 13 s). Citation: R. D. Antia and Cs. Horvath, J. Chromatogr., 435, 1-15 (1988).
12 A Totally Impractical High Temperature Ultrafast Liquid Chromatography (HTUFLC) System injector very small thermal equilibrium delay pump 1ml/min column detector pump 1-14ml/min long delay heating bath ( o C) cooling bath (0 o C)
13 Effect of Temperature on Column Efficiency in HTUFLC o C 80 o C Plate Height ( x10-4 cm ) o C 150 o C u ( cm/s ) Conclusion: Resistance to mass transfer is greatly reduced at elevated column temperature., 5 o C (decanophenone, k =1.3),, 80 o C (dodecanophenone, k =7.39),, 10 o C (tetradecanophenone, k =1.3).
14 Effect of Temperature on Column Dynamics Experimental Conditions a van Deemter Equation Coefficients T( o C) Mobile Phase (% ACN (v/v)) D m x10 4 A x 10 3 (cm /s) b cm B x 10 4 (cm /s) C x 10 3 (s) u opt (cm/s) ± ± ± ± ± ± ± ± ± ± ± ± a Solutes: alkylphenones b Estmated solute diffusion coefficient in the indicated mobile phase at temperature of the calculation based on modified Wilke-Chang equation.
15 A Totally Practical Heating System Cascade Loop Control Column Quasi-Adiabatic Column Environment U.S. Patent Issued Systec/MetalOx
16 References B. Yan, J. Zhao, J.S. Brown, J. Blackwell, and P.W. Carr, High Temperature Ultrafast Liquid Chromatography, Anal. Chem. 7, (000). J.D. Thompson, J.S. Brown, and P.W. Carr, Dependence of Thermal Mismatch Broadening on Column Diameter in High-Speed Liquid Chromatography at Elevated Temperatures, Anal. Chem.73, (001). J.D. Thompson and P.W. Carr, A Study of the Critical Criteria for Analyte Stability in High-Temperature Liquid Chromatography, Anal. Chem. 74, (00). J.D. Thompson and P.W. Carr, High-Speed Liquid Chromatography by Simultaneous Optimization of Temperature and Eluent Composition, Anal. Chem. 74, (00).
17 Theory of High Speed HPLC
18 Rearrangement to Obtain the Guiochon Equation Fundamental Equation # 1 Fundamental Equation # Fundamental Equation # 3 Guiochon Equation L = NH = Nhd p u = νd m dp L t R = ( 1+ k') u tr (1 + k') h = d N D ν m p Knox Equation 1 / 3 B h = Aν + + ν Cν G. Guiochon, Anal. Chem., 1980, 5,
19 Limit 1: C term Knox, Saleem, Guiochon Equation tr N = (1 + k') h d D ν m p Theoretical Limit h Cν as ν Theoretical Limit for t R /N Stokes-Einstein h Cν t R C(1 + ν N D m k') d RT D m = 6nπrη p Result t R ηc(1 + ν N T G. Guiochon, Anal. Chem., 1980, 5, k') d p
20 Limit : A term Practical Limit for h 1/ 3 h Aν t R /N f(a) Kozeny-Carman Permeability and Darcy s Law 4/3 R Ad p (1 + t N = P = D Φ d 1/3 m p u /3 ulη k') Maximum Linear Velocity u max = d p P Lφη max Practical Limit Temperature Dependence (Aν 1/3 > Cν) tr N h Aν ν / 3 (1 + k') L P /3 /3 max η 1/3 T
21 Dependence of t/n on Velocity in the Limit of Exponent of ν x Critical Exponents d p ν x x L x P x η x T x ½ ½ ½ /3 0 /3 - /3 1-1/
22 Relative Viscosity vs. Temperature η/η (5 o C) Water 0 % ACN/Water Methanol Acetonitrile Temperature ( o C)
23 Limit 3: Resolution ') (1 p m R d h D k N t ν + = General Resolution Equation Result ' 1 ' 1 4 k k N R + = α α P h N k tr Φ + = η ') (1 Rearrangement of Darcy s Law m p D hn P d η ν Φ = Retention Time ( ) ' ' k k P h R tr + Φ = α α η Knox-Saleem Equation
24 Dependence of t/n on Optimization Parameters x d p L x P x η x C Limit A limit (ν 1/3 ) 0 /3 - /3 1 Resolution Limit
25 Relative Viscosity vs. Temperature η/η (5 o C) Water 0 % ACN/Water Methanol Acetonitrile Temperature ( o C)
26 Effect of Composition on Viscosity Viscosity of Acetonitrile-Water at 0 o C Viscosity (cp) % Acetontrile
27 Effect of φ, & T on k log k = A/T +B log k = Cφ +B 1.5 log k' log k' / T (K -1 ) φ
28 How Should the Separation Be Done? The same k can be achieved by use of : a. low temperature and organic rich eluent. OR b. high temperature and organic poor eluent. Which allows the faster separation?
29 Is High T Optimal for High Speed in k too high LC? φ (low) φ (high) T (low) low,low low, high T (high) high, low high, high Regions of same k and low viscosity. Where should we work? k too low
30 Effect of φ and T ( at k = 5) on η k (φ,t) 5 % ACN (v/v) 69 T ( o C) 5 η(cp) (φ,t) 0.55 η (T)/η (5 o C) Conditions: k based on butyl benzene on a C 18 Zorbax column.
31 Conclusions 1. To do fast LC, use a WEAK eluent and a HOT column.. Use a highly retentive column so that you can work at lowest possible viscosity!
32 The Importance of Speed in Comprehensive Two-Dimensional HPLC For comprehensive DLC, the speed of the second dimension separation is the rate limiting step in completing the entire D chromatogram. Each first dimension peak must be chromatographed 3-4 times by the second dimension column. 0.5 t rtotal = N ( k' + 1)( k' + ) 1Lc max1 max 1 υ AU Time (min.) Typical Fast 1st Dim. k' max nd Dim. k' max 5 5 N 1 (Plates/column) L c, (cm) u (cm/s) Total Analysis Time (Hrs) 1 1
33 Potential Approaches to Improving the Speed of HPLC Approach Advantage Disadvantage Shorter Columns Works with most equipment, stationary phases Low plate count and resolution Monolithic Columns Ultra-High Pressure LC High Temperature LC Low backpressure High plate counts with small particles Low backpressure, high efficiency at high velocity Narrow-bore columns are not available, high sovent useage Specialized equipment needed, losses in N at high velocity Requires adequate heating, stable phases, stable analytes. High temperature LC is the only approach that allows a significant fraction of the column plate count to be retained as the column linear velocity is increased to values that allow much faster HPLC
34 Schematic of a Complete LC UFHTLC System Standard HP1090 Pump and Autosampler Binary Pump #1 Auto- Injector Eluent Preheater T1 Heating Jacket 1st Dimension Column T T6 Heating Jacket nd Dimension Column T5 Eluent Preheater T4 Sample Loop #1 10-port, -position valve Temperature Controlled.1 mm column Sample Loop # Counter- Current Heat Exchanger T7 UV Detector T3 Eluent Preheater Binary Pump # 5 ml/min pump
35 LC UFHTLC Separation of Ten Triazine Herbicides mau H 3 C H N N N Cl N H N CH 3 Simazine 1 st Dimension Conditions: Column, 50 mm x.1 mm i.d. PBD-ZrO ; Mobile phase, 0/80 ACN/Water; Flow rate, 0.08 ml/min.; Injection volume, 0 µl; Temperature, 40 o C Tim e (m in.) nd Dimension Conditions: Column, 50 mm x.1 mm i.d. PBD-C-ZrO ; Mobile phase, 0/80 ACN/Water; Flow rate, 7.0 ml/min.; Injection volume, 15 µl; Temperature, 150 o C; 1 st dimension sampling frequency, 0.1 Hz Total LC UFHTLC peak capacity = 185 Using a single column, it would take a.5 meter column and 44 hours to generate the same peak capacity
36 Thanks! Ben Yan (ZirChrom). NIH. Carl Sims and Systec, Inc. ZirChrom Separations, Inc.
37 High Throughput Gradient Elution 17 Gradients/Hour. Peak capacity is 70! This speed cannot be done at ambient within the gradient space! Carl Sims Systec Fast Gradient 75 C (17 gradients per hour) Conditions Flow Rate: ml/min Gradient 0-100%B in.5 min. A: 5% THF in Water B: 5% THF in Acetonitrile Column: Zirchrom Diamondbond C18.1 x 50 x 3 micron. System Pressure: 4000 psi System: Waters 790, 487 with microcell Alkyl Phenone Homologs to C Absorbance Time, Minutes 9:58 AM 10:03 AM 10:07 AM 10:1 AM 10:17 AM 10:1 AM 10:6 AM 10:31 AM 10:35 AM
38 Second Dimension Chromatograms mv nd Dimension Time (seconds)
High Speed 2D-HPLC Through the Use of Ultra-Fast High Temperature HPLC as the Second Dimension
High Speed 2D-HPLC Through the Use of Ultra-Fast High Temperature HPLC as the Second Dimension Minnesota Chromatography Forum Spring Symposium Dwight Stoll and Peter W. Carr Department of Chemistry University
More informationAn Overview of the the Development of Stationary Phases for Reversed-Phase Liquid Chromatography
An Overview of the the Development of Stationary Phases for Reversed-Phase Liquid Chromatography Analytical Potential of Stable Phases for Reversed-Phase Liquid Chromatography by Jacek Nawrocki, Jon Thompson,
More informationZirChrom Columns and the Metalox 200-C High Temperature Liquid Chromatography Column Heater - The Future of HPLC has Arrived.
ZirChrom Columns and the Metalox 2-C High Temperature Liquid Chromatography Column Heater - The Future of HPLC has Arrived. Welcome to the fifth issue of ZirChrom's electronic newsletter. In response to
More informationRetention and Selectivity Variables
Applications of Ultra-Stable Phases for HPLC: High Temperature Ultra-Fast Liquid Chromatography and Thermally Tuned Tandem Column (T 3 C) Liquid Chromatography Yun Mao, Merck & Co. Jon Thompson, Peter
More informationEasy Method Transfer from HPLC to RSLC with the Dionex Method Speed-Up Calculator
Technical Note 75 Easy Method Transfer from HPLC to RSLC with the Dionex Method Speed-Up Calculator Introduction The goal of every chromatographic optimization is a method that sufficiently resolves all
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 informationLuna 2.5 µm C18(2)-HST. Advantages of 2.5 µm for increasing the speed of analysis while maintaining high efficiency
Luna 2.5 µm C18(2)-HST Advantages of 2.5 µm for increasing the speed of analysis while maintaining high efficiency Table of Contents Part 1 Theory 1.1 Abstract...3 1.2 Introduction...3 Part 2 Set Up 2.1
More informationHigh-Speed Liquid Chromatography by Simultaneous Optimization of Temperature and Eluent Composition
Anal. Chem. 00, 74, 4150-4159 High-Speed Liquid Chromatography by Simultaneous Optimization of Temperature and Eluent Composition Jonathan D. Thompson and Peter W. Carr* Department of Chemistry, University
More informationWelcome to our E-Seminar: Choosing HPLC Columns for Faster Analysis Smaller and Faster
Welcome to our E-Sear: Choosing HPLC Columns for Faster Analysis Smaller and Faster High Throughput/Fast LC Requires. Short columns 0 mm or shorter Small particle sizes. µm Rapid Resolution or new.8 µm
More informationHigh Speed vs. High Resolution Analyses in HPLC: A Critical Performance Comparison of Column Options Using Poppe and Kinetic Plots
High Speed vs. High Resolution Analyses in HPLC: A Critical Performance Comparison of Column Options Using Poppe and Kinetic Plots Jason C. Link, William E. Barber, William Long, Anne Mack Agilent Technologies
More informationHigh Resolution Fast LC
High Resolution Fast LC Easier Than You Think Rita Steed LC Columns Application Engineer May 9, 2013 What is High Resolution Fast LC? Maintain Resolution with Faster Run Time Increased Resolution with
More informationPractical Applications of Method Translation Using the Agilent Method Translation Tool Rita Steed Inside Application Engineer Agilent Technologies
Practical Applications of Method Translation Using the Agilent Method Translation Tool Rita Steed Inside Application Engineer Agilent Technologies Title Objectives Demonstrate Agilent Method Translation
More informationLECTURE 2. Advanced Separation Science Techniques Present and Future Separation Tools
LECTURE 2 Advanced Separation Science Techniques Present and Future Separation Tools Jack Henion, Ph.D. Emeritus Professor, Analytical Toxicology Cornell University Ithaca, NY 14850 Lecture 2, Page 1 Contents
More informationImproving HPLC Column Selection and System Performance
Improving HPLC Column Selection and System Performance Richard A. Henry Penn State University T410170 sigma-aldrich.com LC Particle Innovation Leads the Way* Particle Size and Architecture (porosity) More
More informationThe Secrets of Rapid HPLC Method Development. Choosing Columns for Rapid Method Development and Short Analysis Times
The Secrets of Rapid HPLC Method Development Choosing Columns for Rapid Method Development and Short Analysis Times Rapid Analysis Is More Than Run Time It is developing a method to meet a goal and developing
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 informationPacked Column for Ultra-Fast Reversed-Phase Liquid Chromatography, TSKgel Super-ODS. Table of Contents
No. 089 SEPARATION REPORT Packed Column for Ultra-Fast Reversed-Phase Liquid Chromatography, TSKgel Super-ODS Table of Contents 1. Introduction 1 2. Column Specification 1 3. Features of Packing Materials
More informationAssay Transfer from HPLC to UPLC for Higher Analysis Throughput
MAY 2005 SEPARATION SCIENCE REDEFINED 31 Assay Transfer from HPLC to UPLC for Higher Analysis Throughput A typical HPLC assay was transferred and optimized for a Waters ACQUITY UPLC system to achieve both
More informationChem 230, Fall, 2014 Homework Set # 3 Short Answer SOLUTIONS
Chem 230, Fall, 2014 Homework Set # 3 Short Answer SOLUTIONS 1. List two advantages of temperature programming in GC. a) Allows separation of solutes with widely varying retention factors in a reasonable
More informationChapter 26: An Introduction to Chromatographic Separations
Chapter 26: An Introduction to Chromatographic Separations Column Chromatography Migration Rates Distribution Contstants Retention Times Selectivity Factor Zone Broadening & Column Efficiency Optimizing
More informationSynthesis and Use of a New Covalently Bonded C18 Modified Carbon Clad Microporous Zirconia for Fast High Temperature Separations
Synthesis and Use of a New Covalently Bonded C18 Modified Carbon Clad Microporous Zirconia for Fast High Temperature Separations by Peter W. Carr, Clayton V. McNeff, Dwight R. Stoll, Danielle R. Hawker,
More informationCHEM340 Tutorial 4: Chromatography
CHEM340 Tutorial 4: Chromatography 1. The data in the table below was obtained from a chromatogram obtained with a 10 cm liquid chromatography column. Under the conditions used, the compound uracil is
More informationImpact of Instrument Dispersion on Performance of HPLC Capillary Columns
Impact of Instrument Dispersion on Performance of HPLC Capillary Columns Wendy Roe, Richard A. Henry, and Hillel Brandes Supelco, Div. of Sigma-Aldrich Bellefonte, PA 16823 USA T413131 sigma-aldrich.com/analytical
More informationCarbonaceous RPLC Stationary Phases Based on Porous Zirconia STABILITY is the Name and SELECTIVITY is the Game Paul T. Jackson
Carbonaceous RPLC Stationary Phases Based on Porous Zirconia STABILITY is the Name and SELECTIVITY is the Game Paul T. Jackson Chemistry Department, St. laf College 1520 St. laf Avenue Northfield, MN 55057
More informationAdvantages and Applications of Revolutionary Superficially Porous Particle Columns in Liquid Chromatography
WHITE PAPER Fast and Efficient HPLC Separations With Superficially Porous Particles Authors D. Guillarme, S. Fekete University of Geneva, University of Lausanne Analytical Pharmaceutical Chemistry Advantages
More information637. Thiamethoxam. HPLC method
637. Thiamethoxam HPLC method CIPAC Collaborative Trial according to CIPAC Information Sheet N o 293 Dr. Sven Adolph Syngenta Crop Protection Münchwilen AG CH-4333 Münchwilen Switzerland May 212 page 1
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 informationAlternatives to Sub-2 µm UHPLC Columns
Alternatives to Sub-2 µm UHPLC Columns J. J. DeStefano, B. E. Boyes, S. Schuster, W. L. Miles, and J. J. Kirkland Advanced Materials Technology, Inc. 3521 Silverside Rd., Quillen Bld., Ste. 1-K Wilmington,
More informationPerformance characteristics of the Agilent 1290 Infinity Quaternary Pump
Performance characteristics of the Agilent 129 Infinity Quaternary Pump Technical Overview Author A.G.Huesgen Agilent Technologies, Inc. Waldbronn, Germany Abstract This Technical Overview presents Proof
More informationDilution(*) Chromatography
WA20264 Poster # 184, HPLC 2002, Montreal, 4-5 June 2002 At-Column Column-Dilution for Preparative Dilution(*) Chromatography Cecilia Mazza, Jie Cavanaugh, Ziling Lu,Tom Sirard,Tom Wheat and Uwe Neue Waters
More informationBetter HPLC Methods Using Temperature Programming
Better HPLC Methods Using Temperature Programming Stephanie J. Marin Brian A. Jones, W. Dale Felix Selerity Technologies, Inc. Salt Lake City, UT www.selerity.com 1 Abstract Temperature programmed liquid
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 informationOn the Advantages and Costs of Two-dimensional HPLC (2D-LC)
On the Advantages and Costs of Two-dimensional HPLC (2D-LC) Georges Guiochon, Krisztìan Horvàth, Jacob Fairchild CoSMoS, Boston, 08/3-5/2009 1 What is Chromatography About? Separating Components of Mixtures
More informationApplication Note Pharmaceutical QA/QC. Agilent Application Solution. Authors. Abstract. Syed Salman Lateef Agilent Technologies, Inc.
Agilent Application Solution Transfer of a USP method for tolazamide from normal phase HPLC to SFC using the Agilent 126 Infinity Hybrid SFC/UHPLC System Improving peak shape and providing wider UV selectivity
More informationCharacteristics and Advantages of Zirconia-Based Stationary Phases for Use in Multi-Dimensional HPLC
Characteristics and Advantages of Zirconia-Based Stationary hases for Use in Multi-Dimensional LC Dwight Stoll and Clayton V. Mceff ZirChrom Separations, Inc. Multi-Dimensional Chromatography Workshop
More informationPerformance evaluation of the Agilent 1290 Infinity 2D-LC Solution for comprehensive two-dimensional liquid chromatography
Performance evaluation of the Agilent 1290 Infinity 2D-LC Solution for comprehensive two-dimensional liquid chromatography Technical Overview 2D-LC Conventional 1D-LC Abstract This Technical Overview presents
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 informationAutomated Switching Between 1D-LC and Comprehensive 2D-LC Analysis
Automated Switching Between D-LC and Comprehensive D-LC Analysis The Agilent 90 Infinity D-LC Solution Technical Overview Author Sonja Krieger Agilent Technologies, Inc. Waldbronn, Germany Abstract This
More informationHPLC Praktikum Skript
HPLC Praktikum Skript Assistants: Gianluca Bartolomeo HCI D330, 3 46 68, bartolomeo@org.chem.ethz.ch Sahar Ghiasikhou HCI E330, 2 29 29, ghiasikhou@org.chem.ethz.ch 1. Introduction In chromatographic techniques,
More informationHPLC. High Performance Liquid Chromatography (HPLC) Harris Chapter 25
High Performance Liquid Chromatography (HPLC) Harris Chapter 25 12/1/2005 Chem 253 - Chapter 25 1 HPLC Separation of nonvolatile or thermally unstable compounds. If the analyte/sample can be found to be
More informationThermo Scientific Accucore XL HPLC Columns. Technical Manual
Thermo Scientific Accucore XL HPLC Columns Technical Manual Thermo Scientific Accucore XL HPLC Columns Based on Core Enhanced Technology using µm solid core particles, Accucore XL HPLC columns allow users
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 informationReversed Phase Solvents
Part 1. General Chromatographic Theory Part 2. verview of HPLC Media Part 3. The Role of the Mobile Phase in Selectivity Part 4. Column Care and Use Reversed Phase Solvents 2 Solvents for RP Chromatography
More informationIsomer Separation on ZirChrom -CARB
Bulletin #34 Isomer Separation on ZirChrom CARB Analytes 6 Ethylbenzene 5 3 mxylene 4 3 4 3 pxylene 4 Xylene 4 6 8 min LC Conditions Column: A: ZirChrom CARB, 5 4.6 mm; Mobile Phase: 5/5 A/B B: Water Flow
More informationComparison of Solid Core HPLC Column Performance: Effect of Particle Diameter
Comparison of Solid Core HPLC Column Performance: Effect of Particle Diameter Luisa Pereira, Thermo Fisher Scientific, Runcorn, Cheshire, UK Technical Note 20755 Key Words Solid core, fused core, superficially
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 information2] The plate height in chromatography is best described as 2
9 Chromatography. General Topics 1] Explain the three major components of the van Deemter equation. Sketch a clearly labeled diagram describing each effect. What is the salient point of the van Deemter
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 informationDeveloping a fast, generic method for rapid resolution LC with quadrupole MS detection. Application Note
Developing a fast, generic method for rapid resolution LC with quadrupole MS detection Combining the Agilent 6140 quadrupole MS and multimode source with the Agilent 1200 Series Rapid Resolution LC system
More informationLC Technical Information
LC Technical Information Method Transfer to Accucore.6 μm Columns Containing solid core particles, which are engineered to a diameter of.6μm and a very narrow particle size distribution; Accucore HPLC
More informationBasic chromatographic parameters and optimization in LC
AM0925 Assignment Basic chromatographic parameters and optimization in LC Introduction This is a computer exercise where you will apply a simulator of reversed phase LC to study the influence of chromatographic
More informationHow to Troubleshoot a Failed System Suitability Test
Volume 34 Number 12, 881 932 Volume 34 Number 12 December 216 www.chromatographyonline.com LCGC NORTH AMERICA How to Troubleshoot a Failed System Suitability Test December 216 Preventing Carryover in GC
More informationEffect of temperature in reversed phase liquid chromatography
Journal of Chromatography A, 1052 (2004) 39 51 Effect of temperature in reversed phase liquid chromatography D. Guillarme, S. Heinisch, J.L. Rocca Laboratoire des Sciences Analytiques, CNRS UMR 5180, Université
More informationNew 5-micron HALO-5 columns based on Fused- Core particle technology boost the performance of HPLC. Compared to other HPLC columns, HALO-5 columns
New 5-micron HALO-5 columns based on Fused- Core particle technology boost the performance of HPLC. Compared to other HPLC columns, HALO-5 columns have: the highest plate number versus any other 5-micron
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 informationMulti-mode Separations Using Zirconiabased Stationary Phases
Multi-mode Separations Using Zirconiabased Stationary Phases PITTC 2010 Dan owlan 1, Bingwen Yan 1, Clayton V. Mceff 1, R.A. Henry 2 1 ZirChrom Separations, Inc. 617 Pierce St., Anoka, M 55303 2 Independent
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 informationA C18 Silica Column With Exceptional Temperature and ph Stability
A C18 lica Column With Exceptional Temperature and ph Stability Brian A. Jones 1, Stephanie J. Marin 1, Jody Clark 1, Nathan L. Porter 1, J. Andreas Lippert 2, and Todd M. Johnson 2 1. Selerity Technologies,
More informationUnderstanding the Use of Temperature Regulation to Optimize Mass Transfer in Fast Gradient Reversed Phase Liquid Chromatography
Understanding the Use of Temperature Regulation to Optimize Mass Transfer in Fast Gradient Reversed Phase Liquid Chromatography Mark J. Hayward Lundbeck Research USA Chemistry : Automation, Profiling,
More informationMethod Development on Next Generation RPLC Supports
Method Development on Next Generation RPLC Supports Dr. Peter W. Carr President Dr. Clayton McNeff Vice President 1-866-STABLE-1 www.zirchrom.com Specialists in High Efficiency, Ultra-Stable Phases for
More informationfor Acclaim Mixed-Mode HILIC-1 Column
for Acclaim Mixed-Mode HILIC-1 Column Product Manual for ACCLAIM Mixed-Mode HILIC-1 Page 1 of 17 Product Manual for ACCLAIM Mixed-Mode HILIC-1 Column 5µm, 4.6 x 250mm, P/N 066844 5µm, 4.6 x 150mm, P/N
More informationPolymer Coated Titania for Analytical and Preparative Reversed-Phase Chromatography
Polymer Coated tania for Analytical and Preparative Reversed-Phase Chromatography EAS 200 BIGWE YA 1, CLAYT V. MCEFF 1, JCHE WIKLER 2 1 ZirChrom Separations, Inc., 617 Pierce Street, Anoka, M 303 2 Sachtleben,
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 informationHPLC. GRATE Chromatography Lab Course. Dr. Johannes Ranke. September 2003
HPLC GRATE Chromatography Lab Course Dr. Johannes Ranke Organisation The groups Start at 9:00 am End at 18:00 pm at the latest Friday, 19th we will finish at 2:00 pm Thursday, 11th: Lecture at 08:15 am
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 informationProduct Bulletin. ACE LC/MS and Rapid Analysis HPLC Columns. HPLC Columns
Product Bulletin HPLC Columns ACE LC/MS and Rapid Analysis HPLC Columns 0 mm, 30 mm, 35 mm and 50 mm column lengths.0,., 3.0, 4.0 and 4.6 mm column diameters Configured for High Sample Throughput Specially
More informationComprehensive 2D-LC Analysis of Chinese Herbal Medicine
Comprehensive 2D-LC Analysis of Chinese Herbal Medicine The Agilent 1290 Infinity 2D-LC Solution Application Note Small Molecule Pharmaceuticals & Generics Author Sonja Krieger Agilent Technologies, Inc.
More informationTransferring HPLC methods from the HP 1090 Series to the Agilent 1100 Series HPLC system
Transferring HPLC methods from the HP 1090 Series to the Agilent 1100 Series HPLC system Technical Note Introduction The HP 1090 Series HPLC system, introduced in 1983, is regarded to be an outstanding
More informationC18 Column. Care & Use Sheet
C18 Column Care & Use Sheet HALO Description HALO C18 is a high-speed, high-performance liquid chromatography column based on a new Fused-CoreTM particle design. The Fused-Core particle provides a thin
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 informationChromatography and other Separation Methods
Chromatography and other Separation Methods Probably the most powerful class of modern analytical methods for analyzing mixture of components---and even for detecting a single component in a complex mixture!
More informationPrep 150 LC System: Considerations for Analytical to Preparative Scaling
Andrew Aubin and Jo-Ann Jablonski Waters Corporation, Milford, MA, USA APPLICATION BENEFITS The Prep 150 LC System is an affordable, highly reliable system for preparative chromatography and is suitable
More informationTransferring Yesterdays Methods to Tomorrows Technology: The Evolution from HPLC to UPLC TM
Transferring Yesterdays Methods to Tomorrows Technology: The Evolution from HPLC to UPLC TM Eric S. Grumbach Thomas E. Wheat Chuck Phoebe Joe Arsenault Jeffrey R. Mazzeo Diane M. Diehl UPLC TECHNOLOGY
More informationIntroduction to Chromatographic Separations (Chapter 1) Many determinations involve separation followed by analysis chromatography electrophoresis
Introduction to Chromatographic Separations (Chapter 1) Many determinations involve separation followed by analysis chromatography electrophoresis Chromatography: sample transported by mobile phase electrostatic
More informationPhenyl-Hexyl. UHPLC Columns. Alternate, complementary selectivity to C18 and C8 bonded phases
Phenyl-Hexyl UHPLC Columns Alternate, complementary selectivity to C8 and C8 bonded phases Particularly recommended for compounds containing aromatic groups Excellent bonded phase stability for durable,
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 informationToo Polar for Reversed Phase What Do You Do?
Too Polar for Reversed Phase What Do You Do? June 20, 2013 Mark Powell Columns and Consumables Technical Support Page 1 C8 or C18 Doesn t Always Do the Job Typical reversed phase conditions involve water/buffer
More informationAnalysis of Beer by Comprehensive 2D-LC with the Agilent 1290 Infinity 2D-LC system
Analysis of Beer by Comprehensive 2D-LC with the Agilent 1290 Infinity 2D-LC system Application Note Food Testing & Agriculture Authors Edgar Naegele Agilent Technologies, Inc. Waldbronn, Germany Keiko
More informationBringing uhplc Performance to the Separation of Peptides
P e p t i d e E S - C 1 8 U H P L C C o l u m n s Bringing uhplc Performance to the Separation of Peptides Compatible with UHPLC and conventional HPLC equipment. Sumpfstr. 3, CH-6300 Zug; Fax: 041 748
More informationThe Effects of Carrier Gas Viscosity and Diffusion on Column Efficiency in Capillary Gas Chromatography
Page 1 of 5 Return The Effects of Carrier Gas Viscosity and Diffusion on Column Efficiency in Capillary Gas Chromatography Stephanye D. Armstrong and Harold M. McNair Department of Chemistry, Virginia
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 informationApplication. Authors. Abstract. Introduction. Hydrocarbon Processing
Fast Analysis Method for Rubber Chemical Antidegradants Using Rapid Resolution Liquid Chromatography (RRLC) Systems with Rapid Resolution High Throughput (RRHT) Columns Application Hydrocarbon Processing
More informationSeparations---Chromatography and Electrophoresis
Separations---Chromatography and Electrophoresis Chromatography--one of most diverse and important analytical methods-- Used initially primarily to purify species With advent of sensitive detectors---now
More informationNexera UC Unified Chromatography
Nexera UC Unified Chromatography The latest addition to the chromatography toolbox Dr. Gesa J. Schad Shimadzu Europa GmbH A brief history of SFC ϒ Late 1800 s: it was found that heavy, non-volatile organic
More informationMethod Transfer between HPLC and UHPLC Instruments Equipment-related challenges and solutions
Method Transfer between HPLC and UHPLC Instruments Equipment-related challenges and solutions Today, ultra-high-performance liquid chromatography (UHPLC) has taken a firm foothold in the analytical laboratory.
More informationPolymer Coated Titania for Analytical and Preparative Reversed-Phase Chromatography
Polymer Coated tania for Analytical and Preparative Reversed-Phase Chromatography Pittcon 2005 CLAYT V. MCEFF 1, BIGWE YA 1, JCHE WIKLER 2 1 ZirChrom Separations, Inc., 617 Pierce Street, Anoka, M 55303
More informationInertSustainSwift C8
Physical Properties Silica Particle Size Surface Area Pore Size Pore Volume Bonded Phase End-capping Carbon Loading ph Range USP Code :ES (Evolved Surface) Silica Gel :.9 μm, μm, μm :00 m /g :00 Å (0 nm)
More informationLC III: HPLC. Originally referred to as High-Pressure Liquid Chromatography. Now more commonly called High Performance Liquid Chromatography
LC III: HPLC What is HPLC? Originally referred to as High-Pressure Liquid Chromatography Now more commonly called High Performance Liquid Chromatography In general: The instrument controlled version of
More informationAnalysis of DNPH-derivatized Aldehydes and Ketones using the Agilent 1220 Infinity LC System with Diode Array Detector
Analysis of DNPH-derivatized Aldehydes and Ketones using the Agilent Infinity LC System with Diode Array Detector Application Note Author Sonja Schneider Agilent Technologies, Inc. Waldbronn, Germany [mau]
More informationXiqin Yang, Jun Dai, Peter W. Carr* Chemistry Department University of Minnesota
Comparison of the Reversed-Phase and Ion- Exchange Contributions to Retention on Polybutadiene Coated Zirconia and Octadecyl Silane Bonded Silica Phases Xiqin Yang, Jun Dai, Peter W. Carr* Chemistry Department
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 informationHigh Performance Liquid Chromatography
Updated: 3 November 2014 Print version High Performance Liquid Chromatography David Reckhow CEE 772 #18 1 HPLC System David Reckhow CEE 772 #18 2 Instrument Basics PUMP INJECTION POINT DETECTOR COLUMN
More informationHigh Performance Liquid Chromatography
Updated: 3 November 2014 Print version High Performance Liquid Chromatography David Reckhow CEE 772 #18 1 HPLC System David Reckhow CEE 772 #18 2 1 Instrument Basics PUMP INJECTION POINT DETECTOR COLUMN
More informationChanging the way you think about HPLC
3521 Silverside Road Suite 1-K Quillen Building Wilmington, DE 19810 USA info@advanced-materials-tech.com Changing the way you think about HPLC Changing the way you think about HPLC New 5-micron HALO-5
More informationLC and LC/MS Column Selection Flow Chart
LC and LC/MS Column Selection Flow Chart To use the column selection diagram below, simply follow the path for your analyte and mobile phase. At the far right, follow your final column selection to the
More informationQuantification of growth promoters olaquindox and carbadox in animal feedstuff with the Agilent 1260 Infinity Binary LC system with UV detection
Quantification of growth promoters olaquindox and carbadox in animal feedstuff with the Agilent 126 Infinity Binary LC system with UV detection Application Note Food Author Srividya Kailasam Agilent Technologies,
More informationM > ACN > > THF
Method Development in HPLC Dr. Amitha Hewavitharana School of Pharmacy University of Queensland Method Development in HPLC References: D A Skoog Principles of instrumental analysis, 3 rd Edition Chapters
More informationThe Agilent InfinityLab 2D-LC Solution with Active Solvent Modulation
Technical Overview The Agilent InfinityLab D-LC Solution with Active Solvent Modulation Achieving Improved Resolution and Sensitivity for Challenging Combinations of Separation Conditions Author Sonja
More informationChromatographic Methods: Basics, Advanced HPLC Methods
Chromatographic Methods: Basics, Advanced HPLC Methods Hendrik Küpper, Advanced Course on Bioinorganic Chemistry & Biophysics of Plants, summer semester 2018 Chromatography: Basics Chromatography a physical
More informationBiochemistry. Biochemical Techniques HPLC
Description of Module Subject Name Paper Name 12 Module Name/Title 13 1. Objectives 1.1. To understand the basic concept and principle of 1.2. To understand the components and techniques of 1.3. To know
More information