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HPLC Channel
The Theory Of HPLC

Introduction
  1. Aims and Objectives
  2. Origins of Liquid Chromatography
  3. Why Choose Liquid Chromatography
  4. Suitable Samples for HPLC
  5. Comparison with Gas Chromatography
  6. Typical HPLC Data
  7. Chromatography Separation Mechanisms
  8. The Liquid Chromatograph
  9. The Liquid Chromatographic Process
  10. The Chromatogram
  11. Modes of Chromatography
  12. Assessment
Chromatographic Parameters
  1. Aims and Objectives
  2. Resolution
  3. The Resolution Equation
  4. Retention (Capacity) Factor
  5. How to change Retention (Capacity) Factor
  6. Effect of Retention Factor on Resolution
  7. Selectivity (Separation Factor)
  8. How to change Selectivity (Separation Factor)
  9. Effect of Selectivity on Resolution
  10. Efficiency
  11. How to change Efficiency
  12. Effect of Efficiency on Resolution
  13. Resolution - Real Life Examples
  14. Peak Asymmetry
  15. Assessment
Band Broadening
  1. Aims and Objectives
  2. Band Broadening - the Van Deemter Equation
  3. Band Broadening - Eddy Diffusion
  4. Longitudinal Diffusion
  5. Band Broadening - Mass Transfer
  6. Optimising Flow Rate
  7. Optimising Particle Size
  8. Minimising System Volume
  9. Assessment
Column chemistry
  1. Aims and Objectives
  2. Silica as a Packing Material
  3. Chemically Bonded Phases
  4. Surface Treatment - End capping
  5. Reversed Phase Stationary Phases
  6. Silanol and Separation
  7. Water Wettable Phases
  8. Aq Type Stationary Phases
  9. Polar Embedded Phases
  10. Working at Low pH
  11. Working at High pH
  12. Other Stationary Phase Types
  13. Column Test Probes
  14. Column Characterisation
  15. Assessment
Reverse phase (partition) chromatography
  1. Aims and Objectives
  2. Mechanism of Reversed Phase HPLC
  3. Applications of Reversed Phase HPLC
  4. Analyte Retention in Reversed Phase HPLC
  5. Retention Order in Reversed Phase HPLC
  6. Reversed Phase Mobile phase Solvents (a)
  7. Reversed Phase Mobile phase Solvents (b)
  8. Mobile phase strength and retention
  9. Changing the Organic Modifier
  10. Eluotropic Series
  11. Selecting Reverse Phase Columns
  12. Reverse Phase HPLC of Ionisable Samples
  13. Analyte Ionisation
  14. Controlling Extent of Ionisation
  15. pH vs Retention in Reverse Phase HPLC
  16. Basic Analytes & Ion Suppression
  17. Buffers for Reverse Phase HPLC
  18. Buffer Concentration
  19. Getting Started with Ionisable Compounds
  20. Assessment
Ion-Pair Chromatography
  1. Aims and Objectives
  2. Ion Pair Chromatography - Fundamental Mechanism
  3. Ion Pair Chromatography - Reagents
  4. Ion Pair Chromatography - Retention & Selectivity
  5. Important Parameters in Ion Pair Chromatography
  6. Optimising Ion Pair Concentration
  7. Applications and Ion Pairing for LC-MS
  8. Practical Ion Pair Chromatography
  9. Getting Started with Ion Pair HPLC
  10. Assessment
Normal phase (absorption) chromatography
  1. Aims and Objectives
  2. Mechanism of Normal Phase Chromatography
  3. Applications of Normal Phase Chromatography
  4. Retention and Selectivity Stationary Phases
  5. Stationary Phases for Normal Phase HPLC
  6. Typical Mobile Phases
  7. Controlling Retention
  8. Mobile Phase Optimisation
  9. Problems with Water in the Mobile Phase
  10. Getting Started with Normal Phase HPLC
  11. Assessment
Gradient HPLC
  1. Aims and Objectives
  2. Isocratic HPLC Analysis
  3. Gradient HPLC Analysis
  4. Gradient Elution Parameters
  5. Gradient Elution Principles
  6. Peak Shape in Gradient HPLC
  7. Scouting Gradients
  8. Gradient Steepness
  9. Optimising Gradient Analyses
  10. Practical Gradient HPLC
  11. Estimating Gradient Parameters
  12. Assessment
Quantitative and Qualitative HPLC
  1. Aims and Objectives
  2. Qualitative Analysis Overview
  3. Peak Identification and Assignment
  4. Sample Spiking
  5. Spectral Peak Identification
  6. Quantitative Analysis Overview
  7. Chromatographic Requirements
  8. Peak Integration
  9. Peak Height or Peak Area
  10. Principles of Quantitative Analysis
  11. Area / Height %
  12. External Standard Quantitation
  13. Calibration Curve
  14. External Standard Multi-level Calibration
  15. Internal Standard Analysis
  16. Assessment
*New* Fast HPLC
  1. Aims and Objectives
  2. Introduction
  3. Separation Efficiency
  4. Back Pressure and Particle Size
  5. Superficially Porous Materials
  6. Increasing Resolution via Improved Efficiency
  7. Speeding up HPLC Separations
  8. HPLC Method Conversion
  9. Transferring Methods - Gradient Elution Considerations
  10. Transferring Methods - Calculating Gradient Conditions
  11. Transferring Methods - Quick Method Transfer Tools
  12. Efficiency
  13. How to Change Efficiency
  14. Effects of Efficiency (N) on Resolution
  15. Van Deemter Plots
  16. Eddy Diffusion (The A term)
  17. Longitudinal Diffusion (The B Term)
  18. Mass Transfer (The C Term)
  19. Packing Material - Particle Size
  20. Packing Material - Particle Size Distribution
  21. Column Internal Diameter and Linear
  22. Packing Material - Morphology
  23. Reduced Diameter / Sub 2µm Particles
  24. Superficially Porous Particles
  25. Reduced Diameter Non-Porous Particles
  26. Temperature
  27. Kinetic Plots
  28. Poppe Plots - Use and Description
  29. Poppe Plots - Parameters
  30. Impedance Plots - Use and Description
  31. Impedance Plots –Parameters
  32. Thermal Considerations
  33. Power Plots
  34. Assessment
Instrumentation of HPLC

Mobile Phase Considerations
  1. The Mobile Phase - Introduction
  2. The Liquid Chromatographic Process
  3. Analyte Retention Processes
  4. The Partition Coefficient
  5. Retention and Selectivity
  6. Solvent Type and Selectivity
  7. Optimising Selectivity using Retention
  8. Gradient Elution
  9. Mobile Phase Delivery
  10. Solvent Miscibility
  11. UV Cut Off
  12. Other Solvent Considerations
  13. Mobile Phase Preparation
  14. Degassing
  15. Outlet Degassing
  16. Degassing / Column Damage
  17. Methods of Degassing
  18. Helium Sparging
  19. Vacuum Degassing
  20. Assessment
HPLC Solvent Pumping Systems
  1. Pumps - Introduction
  2. Simple Pumping Systems
  3. Disadvantages of Single Piston Pumps
  4. Dual Piston Reciprocating Pumps
  5. Mixing Solvents, Binary Pumps
  6. Mixing Solvents, Quaternary Pumps
  7. Mixing Solvents, Ternary Pumps
  8. Check Valves
  9. Pistons
  10. Pulse Dampers
  11. Purge Valves
  12. Flushing, Gradients and Gradient Dwell Time
  13. Experimental Determination of System Dwell
  14. Troubleshooting - High Back Pressure
  15. Troubleshooting - Check Valves and Pistons
  16. Troubleshooting - Solvent Mixing Issues
  17. Preparative Pumps
  18. Capillary (Low Volume) Pumps
  19. Calibration and Testing
  20. Assessment
Autosamplers
  1. Introduction
  2. Injection Valves
  3. Injection Valve Anatomy
  4. Manual Injection Systems
  5. Manual Injection Complete and Partial Loop Filling
  6. Autosamplers
  7. Pull to Fill Auto Samplers
  8. Push to Fill Auto Samplers
  9. Integral Loop Auto Samplers
  10. Autosampler Contamination
  11. Autosampler Contamination
  12. Assessment
Detectors
  1. Detectors for HPLC - Introduction
  2. General Terms and Concepts
  3. Limit of Detection / Quantification (LOD / LOQ)
  4. UV - Vis Detectors, The Flow Cell
  5. UV - Vis Detectors, Quantitation
  6. External Standard Quantitation
  7. Calibration Curve
  8. UV - Vis Detectors, UV absorbance
  9. Variable Wavelength UV - Vis Detectors
  10. UV Detectors - Diode Array Detectors
  11. UV Detectors - DAD Spectra
  12. DAD Detectors - Bandwidth
  13. DAD Detectors - Slit Width
  14. DAD Detectors - Response time
  15. DAD Detectors - Reference Wavelength
  16. Choosing Sample and Reference Settings
  17. DAD - Peak Supression
  18. Fluorescence Detectors
  19. Fluorescence Detectors - Principles
  20. Fluorescence Detectors - Excitation & Emission
  21. Refractive Index Detectors - Instrumentation
  22. Refractive Index Detectors - Principles
  23. Assessment
GC Channel
Theory and Instrumentation of GC

Introduction
  1. Aims and Objectives
  2. Origins of Gas Chromatography
  3. Why choose Gas Chromatography (a)
  4. Why choose Gas Chromatography (b)
  5. Gas Chromatography Separation Mechanism
  6. The Gas Chromatograph
  7. The Chromatogram
  8. GC Advantages and Disadvantages
  9. Typical GC Applications
  10. Assessment
Chromatographic Parameters
  1. Aims and Objectives
  2. Chromatographic Resolution (Rs)
  3. The Resolution Equation
  4. Retention Factor (k)
  5. How to change Retention Factor (k)
  6. Effect of Retention Factor on Resolution
  7. Selectivity (Separation) Factor
  8. How to Change Selectivity
  9. Effect of Separation Factor on Resolution
  10. Efficiency
  11. How to change Efficiency
  12. Effect of Efficiency on Resolution
  13. Resolution - Real Life Examples
  14. Resolution - Real Life Examples [Q1]
  15. Resolution - Real Life Examples [Q2]
  16. Resolution - Real Life Examples [Q3]
  17. Peak Asymmetry
  18. Assessment
Band Broadening
  1. Aims and Objectives
  2. Importance of Efficiency in GC Separations
  3. Van Deemter and Golay Equations
  4. Eddy Diffusion
  5. Longitudinal Diffusion
  6. Stationary Phase Mass Transfer
  7. Mobile Phase Mass Transfer
  8. Stationary Phase Film Thickness Effects
  9. Column Internal Diameter Effects
  10. Carrier Gas Flow Rate Effects
  11. Effect of Carrier Gas Type on Efficiency
  12. Assessment
Gas Supply and Pressure Control
  1. Aims and Objectives
  2. Gases required for GC
  3. Gas Supply Management
  4. Quality of Gas Supply
  5. Gas Generators - Using Hydrogen in the Lab
  6. Manual Pressure Control
  7. Electronic Pressure Control
  8. Pressure / Flow Programming
  9. Assessment
Sampling Techniques
  1. Aims and Objectives
  2. Sampling Techniques Overview
  3. Manual Injection
  4. Cold Needle Technique
  5. Hot Needle Technique
  6. Air Gap Techniques
  7. Solvent Flush Technique
  8. Automatic Liquid Sampling (Autosamplers)
  9. Gas Sampling Devices
  10. Purge and Trap Autosamplers (a)
  11. Purge and Trap Adsorbents (b)
  12. Thermal Desorption Autosamplers (TD)
  13. Two Stage Thermal Desorption
  14. Two Stage Desorption with Cold Trapping
  15. Thermal Desorption - Important Parameters
  16. TD Sorbent Selection and Applications
  17. Solid Phase Microextraction (SPME)
  18. Important SPME Parameters
  19. Headspace Sampling (HS)
  20. Headspace Autosamplers
  21. Headspace Calibration and Quantitation
  22. Assessment
Sample Introduction
  1. Aims and Objectives
  2. GC Inlet Systems
  3. Spilt / Splitless Inlet
  4. Spilt Injection
  5. Setting the Split Ratio
  6. Sample Discrimination
  7. Injection Volume
  8. Optimising Injection Volume
  9. Split Injection - Experiments [Q1]
  10. Split Injection - Experiments [Q2]
  11. Split Injection - Experiments [Q3]
  12. Split Injection - Experiments [Q4]
  13. Spiltless Injection
  14. Optimising Splitless Injection - Purging the inlet
  15. Optimising Splitless Injection - Analyte Focussing
  16. Optimising Splitless Injection - Solvent Choice
  17. Splitless Injection - Experiments [Q1]
  18. Splitless Injection - Experiments [Q2]
  19. Splitless Injection - Experiments [Q3]
  20. Choosing an Inlet Temperature
  21. Liners for Split / Splitless injection
  22. Septa for Split / Splitless injection
  23. Septa Problems
  24. Cool-on column (COC) Inlet
  25. Optimising COC injection
  26. Use of Retention Gaps for Cool-on-Column injection
  27. Programmed Thermal Vaporising (PTV) Inlets
  28. PTV Sample Flow Rate
  29. PTV Liner Type and Packing
  30. PTV Flow and Temperature
  31. PTV Solvent Elimination
  32. Direct (Packed Column) Inlets
  33. Assessment
GC Columns
  1. Aims and Objectives
  2. Open Tubular Capillary Columns
  3. Comparison of Packed and Capillary GC columns
  4. Chemistry Review - Analyte & Stationary Phase Polarity
  5. Electronegativity
  6. Dispersive Interactions
  7. Dipole Interactions
  8. Hydrogen Bonding
  9. Stationary Phases - Polysiloxanes
  10. Polysiloxane Classifications
  11. Stationary Phases Polyethylene Glycols
  12. Stationary Phase Selection
  13. Phase selection Dispersive Phases
  14. Dispersive Interactions and Polarity
  15. Dipole Interactions and Hydrogen Bonding
  16. Stationary Phase Selection PLOT Columns
  17. Stationary Phase Selection Summary
  18. Stationary Phases for Packed Column GC
  19. Column Dimensions - Length
  20. Column Dimensions - Internal Diameter
  21. Column Dimensions - Film Thickness (df)
  22. Phase Ratio (ß)
  23. Carrier Gas Flow Rate
  24. Column Bleed
  25. Column Installation & Conditioning
  26. Exercises in Column Selection [Q1]
  27. Exercises in Column Selection [Q2]
  28. Assessment
GC Temperature Programming
  1. Aims & Objectives
  2. The Role of Temperature in GC Separations
  3. Isothermal and Gradient Temperature GC
  4. Theory of Temperature Programmed GC
  5. Using and Developing Temperature Programs
  6. Scouting Gradients
  7. Predicting Isothermal Conditions
  8. Optimising Isothermal Conditions
  9. Initial Temperature and Hold Time
  10. Adjusting the Ramp Rate
  11. Final Temperature and Time
  12. Assessment
GC Detectors
  1. Aims and Objectives
  2. GC Detectors Overview
  3. GC Detectors - Characteristics (a)
  4. GC Detectors - Characteristics (b)
  5. The Flame Ionisation Detector
  6. FID - Operating and Optimising
  7. FID - Uses and Performance
  8. The Nitrogen Phosphorous Detector (NPD)
  9. NPD - Operating and Optimising
  10. NPD - Uses and Performance
  11. The Electron Capture Detector (ECD)
  12. ECD - Operating and Optimising
  13. ECD - Uses and Performance
  14. The Thermal Conductivity Detector (TCD)
  15. TCD - Operating and Optimising
  16. TCD - Uses and Performance
  17. Other GC Detectors (a)
  18. Other GC Detectors (b)
  19. Assessment
MS Channel
Fundamental LC-MS

Introduction
  1. Learning Aims & Objectives
  2. Definitions
  3. Instrument Fundamentals
  4. Process
  5. Why and when to use LC/MS
  6. Ionisation Overview
  7. Ionisation Atmospheric Pressure Ionisation (API)
  8. Ionisation Electrospray Ionisation (ESI)
  9. Ionisation Atmospheric Pressure Chemical Ionisation (APCI)
  10. Ionisation Atmospheric Pressure Photo Ionisation (APPI)
  11. Mass analysers
  12. Quadrupole
  13. Mass analysers Time-of-flight
  14. Mass analysers Ion Trap Mass Analyser
  15. Tandem mass spectrometry (MS/MS)
  16. Detectors
  17. Applications
  18. Assessment
Electrospray Ionisation Theory
  1. Learning Aims & Objectives
  2. Introduction
  3. Suitable analytes for ES
  4. Production of charged droplets at the capillary tip.
  5. Formation of the Taylor cone
  6. Nebulisation Overview
  7. Nebulisation Applied Potential Difference
  8. Spray Breakdown and Discharge
  9. Electrospray Production - Pneumatic Assistance
  10. Electrospray Production - Eluent Flow Rate
  11. Electrospray Production - Surface Tension
  12. Electrospray Production - Ionic Strength
  13. Electrospray Ionisation Droplet Desolvation
  14. Droplet desolvation and Jet fission
  15. Gas Phase Ions The Dole Model
  16. Gas Phase Ions Iribarne / Thompson Model
  17. Gas Phase Ions Experimental Factors
  18. Gas Phase Ions Charge to Droplet Radius Ratio
  19. Gas Phase Ions Energy Barriers
  20. Gas Phase Ions Mass / Charge Ratio
  21. Quantitative aspects Sensitivity constants
  22. Sensitivity constants - Droplet Radius
  23. Droplet Surface Activity
  24. Ion Suppression Principles
  25. Ion Suppression - Competing Ions
  26. Ion Suppression in Practice
  27. Mass Dependence of the Ion Signal
  28. Optimsing the Sprayer Position
  29. Solute Changes in Evaporating Droplets
  30. pH vs Ion Intensity
  31. Assessment
Electrospray Ionisation – Instrumentation
  1. Learning Aims & Objectives
  2. Electrospray source design
  3. Electrospray capillary design
  4. Source Heating (Drying Gas)
  5. T-Piece designs
  6. Optimsing Sprayer / Sampling Plate Configuration
  7. Orthogonal Spray Source Designs
  8. Cluster Ion Sampling
  9. Curtain Gas Systems to Prevent Cluster Ions
  10. Dielectric Capillaries to Prevent Cluster Ions
  11. Source Cleaning
  12. Ion Optics Ring Electrodes
  13. Ion Optics Ion Bridges
  14. Collision Induced Dissociation
  15. Effect of Nozzle Skimmer Voltages
  16. Collision Induced Dissociation Probes
  17. E-Lab
  18. Assessment
Mass Analyzers
  1. Learning Aims & Objectives
  2. Introduction to Mass Analysis
  3. Topic Overview
  4. Terms and Definitions
  5. Quadrupole Mass Analyser Introduction
  6. Quadrupole Rods
  7. Quadrupole Voltages
  8. Quadrupole Electrostatic Fields
  9. Quadrupole Ion Trajectroies
  10. Equations of Ion Motion
  11. Quadrupole Mass Analysers - Mass Gain & Offset
  12. Quadrupole - Resolution and Sensitivity
  13. Quadrupole Data Acquisition Modes
  14. Quadrupole Scan vs. Selected Ion Modes
  15. RF only Ion Bridges
  16. Quadrupole - Mass Accuracy
  17. Quadrupole - Performance Limitations
  18. Quadrupole - Scanning Speeds
  19. Time-of- Flight (TOF) Mass Analysers
  20. TOF - Equations of Motion
  21. TOF - Resolution
  22. TOF - Issues with High Mass Resolution
  23. TOF - The Reflectron
  24. TOF - Increased Resolution using Reflectrons
  25. TOF - Performance Limitations
  26. Orthogonally Accelerated TOF Instruments
  27. oaTOF Pusher Electrode
  28. oaTOF Pusher Pulse Rate
  29. oaTOF Deflecting Voltage
  30. oaTOF Interfacing Details
  31. oaTOF Mass Resolution and Accuracy
  32. oaTOF Dynamic Range
  33. Ion Trap Mass Analysers - Introduction
  34. Ion Trap Equations of Ion Motion
  35. Ion Trap Stability Diagram
  36. Ion Trap Space Charge Effects
  37. Ion Trap Ion Manipulation
  38. Ion Trap Scanning Experiments
  39. Ion Trap Other Ion Experiments
  40. Ion Trap Mass Resolution and Accuracy
  41. Magnetic Sector Mass Analysers - Introduction
  42. Magnetic Sector Equations of Ion Motion
  43. Magnetic Sector Data Acqusition Modes
  44. Electrostatic Analysers
  45. Double Focussing Instruments
  46. Magnetic Sector Ion Optics
  47. Magnetic Sector Performance Limitations
  48. Magnetic Sector Mass Resolution and Accuracy
  49. Magnetic Sector Performance figures
  50. Mass analysers Selection
  51. Introduction to Tandem Mass Spectrometry
  52. Assessment
Atmospheric Pressure Chemical Ionisation (APCI)
  1. Learning Aims & Objectives
  2. Introduction
  3. Interface Overview
  4. Suitable Samples for APCI
  5. APCI Interfacing Details
  6. APCI Nebuliser Types APCI Nebuliser Types
  7. APCI Nebuliser Gas Flow
  8. APCI Interfacing Details
  9. APCI Analyte Ion Declustering
  10. APCI Ionisation - Mechanisms
  11. APCI Proton Affinity
  12. APCI Ionisation - Negative Ion Mode
  13. APCI Ionisation – Positive Ion Mode
  14. APCI Gas Phase Reactant Ions
  15. APCI Ionisation – Negative Ion Mode
  16. APCI Reagent Gas Formation
  17. Eluent Additives in APCI
  18. Signal Suppression by Additives in APCI
  19. Alternative APCI Charging Mechanisms
  20. APCI Source Parameter Optimisation
  21. Comparison of ESI/APCI ionisation techniques
  22. E-Lab
  23. Assessment
Atmospheric Pressure Photoionisation (APPI)
  1. Learning Aims & Objectives
  2. APPI Introduction
  3. APPI Instrumentation
  4. Suitable Samples for APPI
  5. APPI Interfacing Details
  6. APPI Eluent flow rates
  7. APPI Dual mode of operation
  8. APPI Ionisation Mechanisms
  9. APPI Positive ion mode
  10. APPI Negative ion mode
  11. Ion sampling and transfer in APPI interfaces
  12. APPI - Analyte Ion Declustering
  13. APPI Source Parameter Optimisation
  14. E-Lab
  15. Assessment
*NEW* Solvents, Buffers and Additives
  1. Learning Aims & Objectives
  2. Introduction
  3. LC-API Compatibility
  4. ESI Solution Chemistry
  5. ESI Eluent Solvent
  6. Solvent Viscosity in ESI
  7. Organic Modifiers in ESI
  8. Eluent Solvent – Positive ESI
  9. Eluent pH – Positive ESI
  10. Eluent pH – Negative ESI
  11. ESI Positive/Negative Ion Mode
  12. ESI Reagents for pH Control
  13. Sources for non-Volatile Systems
  14. Z Spray and non-Volatile Systems
  15. ESI Test Compound Infusion
  16. ESI Buffer Choice and Concentration
  17. ESI Buffer Systems
  18. ESI Ion Pair Reagents – Overview
  19. ESI Ion Pair Reagents – Considerations
  20. Cationisation vs. Anionisation in ESI
  21. Pre and Post Column Addition in ESI
  22. Post Column Addition in ESI
  23. ESI Adduct Formation
  24. ESI Considerations
  25. APCI Solvent Choice – Overview
  26. APCI Solvent Choice –Mechanisms
  27. Buffers for APCI – Overview
  28. Buffers for APCI – Concentration
  29. Ion pairing reagents in APCI
  30. APCI Considerations
  31. E-Lab
  32. Assessment
*NEW* Vacuum Systems
  1. Learning Aims & Objectives
  2. Introduction
  3. Vacuum Systems
  4. Mean Free Path
  5. Rotary Pumps
  6. Foreline Pumps
  7. Turbomolecular Pumps
  8. Diffusion Pumps
  9. Vacuum and Flow Rate Incompatibility
  10. Establishing Vacuum and Transmission
  11. The Sampling Orifice Plate
  12. Nozzle Skimmer Region – Analyte Enrichment
  13. Second Pumping Stage
  14. Molecular Beam Theory
  15. Practical Implication of the Skimmer Position
  16. Vacuum System Troubleshooting & Maintenance
  17. Vacuum Leaks
  18. Foreline Pumps – Gas Ballasting
  19. Foreline Pumps – Exhaust Filters
  20. Foreline Pumps – Rotary Pump Oil
  21. High Vacuum Pumps
  22. Assessment
*NEW* Flow Rates and Flow Splitting
  1. Learning Aims & Objectives
  2. Introduction
  3. Interface Similarities
  4. The ESI Interface
  5. The APCI Interface
  6. Flow Rates for APCI
  7. Flow Rates for ESI
  8. Flow Rates for Pneumatically Assisted ESI
  9. Micro and Nanoflow ESI
  10. Flow Rate Incompatibility – Reasons
  11. Flow Rate Incompatibility – Solutions
  12. Flow Splitting – Overview
  13. Flow Splitting – Adjustable Flow Splitters
  14. Flow Splitting – Practicalities
  15. MS Detector
  16. Columns – Internal Diameter
  17. Columns – Low Flow Rates
  18. Capillary LC systems – Overview
  19. Capillary LC Systems – Practicalities
  20. Column Selection
  21. E-lab
  22. Assessment
MS Interpretation

*NEW* General Interpretation Strategies
  1. Learning Aims & Objectives
  2. Introduction
  3. Mass to Charge Ratio
  4. Mass Resolution
  5. High Mass Resolution
  6. Mass Accuracy
  7. High Mass Accuracy
  8. Mass Range
  9. Multiply Charged Ions
  10. Spectral Features
  11. Isotopic abundances
  12. High Mass Region – Brominated Sample
  13. High Mass Region – Chlorinated Sample
  14. High Mass Region – Dichlorinated Sample
  15. The Nitrogen Rule
  16. Interpretation Strategy
  17. Fragmentation in API
  18. Rings and Unsaturations
  19. Number of Carbons
  20. Postulating a Molecular Formula
  21. Cleavages –Ion Abundance
  22. Cleavages –Simple Mechanisms
  23. Electrospray Ionisation
  24. ESI Considerations
  25. APCI Considerations
  26. APPI Considerations
  27. LC-MS Structural Information Modes
  28. MS/MS Overview
  29. MS/MS Experiments
  30. Product Ion Scanning - Overview
  31. Product Ion Scanning – Application
  32. Precursor Ion Scanning - Overview
  33. Precursor Ion Scanning – Application
  34. Constant Neutral Loss Scanning - Overview
  35. Constant Neutral Loss Scanning – Application
  36. Single/Multiple Reaction Monitoring - Overview
  37. Single/Multiple Reaction Monitoring - Apllication
  38. MS/MS Quantitative Considerations
  39. E-Lab Part 1 - Preliminary condiderations
  40. E-Lab Part 1 - Molecular Weight
  41. Assessment
Fundamental GC-MS

Introduction
  1. Learning Aims and Objectives
  2. Definitions
  3. Instrument Fundamentals GC
  4. Instrument Fundamentals MS
  5. GC-MS Process
  6. Why and when to use GC-MS
  7. Coupling GC to MS systems
  8. Ionisation Overview
  9. Ionisation - Electron Impact (EI)
  10. Ionisation - Chemical Ionisation (CI)
  11. Ionisation - Suitable samples for GC-MS
  12. Mass analysers – Overview
  13. Mass analysers – Quadrupole
  14. Mass analysers – Time of flight (TOF)
  15. Mass analysers – Ion trap
  16. Mass analysers – Magnetic sector
  17. Tandem mass Spectrometry (MS-MS)
  18. Detectors
  19. Applications
  20. Assessment
GC Considerations
  1. Learning Aims and Objectives
  2. Introduction
  3. Carrier gas
  4. Sample introduction
  5. Split Injection – Overview
  6. Split Injection– Setting Split Ratio
  7. Split Injection – Sample Discrimination
  8. Split Injection – Injection Volume
  9. Splitless Injection – Overview
  10. Splitless Injection – Purging the Inlet
  11. Split Injection – Analyte Focusing
  12. Split Injection – Solvent Choice
  13. PTV inlets
  14. Headspace sampling
  15. Headspace Autosamplers
  16. Headspace Analysis Important Parameters
  17. Columns
  18. Stationary phases
  19. GC-MS column selection
  20. Fittings
  21. Guard columns (Retention Gap)
  22. Air leaks
  23. Ferrules
  24. Ferrules – Practicalities
  25. Septum Overview
  26. Septum – Selection
  27. Septum Considerations
  28. Contamination
  29. Assessment
*New* GC -MS Interfaces
  1. Learning Aims & Objectives
  2. Introduction
  3. Coupling GC to MS Detectors - Jet Separator
  4. Coupling GC to MS Detectors - Direct Interface
  5. Column Diameter
  6. Interface-column coupling I
  7. Interface-column coupling II
  8. Interface-column coupling III
  9. Interface-column coupling IV
  10. Assessment
Sample Preparation Channel
Solid Phase Extraction

Molecular Properties
  1. Learning Aims & Objectives
  2. Functional groups
  3. Molecular Properties
  4. Functional group interactions
  5. Hydrophobic or Non-Polar Groups
  6. Hydrophobic Interactions - Solubility
  7. Hydrophobic Interactions - Sorbents
  8. Polar Groups
  9. Polar Interactions - Solubility
  10. Polar Interactions - Sorbents
  11. Ionic groups
  12. Ionic groups - pH
  13. Ionic groups - Ka
  14. Ionic groups - pKa
  15. Ionic groups -Ionic strength
  16. Ionic Interactions - Solubility
  17. Ionic Interactions - Sorbents
  18. Chelating Groups
  19. Chelating Groups - Chelating Interactions - Solubility
  20. Chelating Groups - Chelating Interactions - Sorbents
  21. Protein Binding
  22. Assessment
SPE Overview
  1. Learning Aims & Objectives
  2. Solid Phase Extraction - Overview
  3. SPE Terminology
  4. SPE Sorbent Physical Properties l
  5. SPE Sorbent Substrates
  6. SPE Sorbent Surface Chemical Nature
  7. SPE Sorbent Surface Chemical Nature
  8. Choosing Sorbent Mass
  9. Protocol Steps in SPE
  10. SPE Sample Pretreatment
  11. SPE Column Conditioning
  12. SPE Column Equilibration
  13. Sample Loading
  14. Column Washing
  15. Analyte Elution
  16. Pharmaceutical SPE
  17. Assessment
SPE Mechanisms
  1. Learning Aims & Objectives
  2. Non-Polar SPE
  3. Non-Polar Sample Pretreatment
  4. Equilibration and Sample Loading
  5. Non-Polar Sorbent Washing and Analyte Elution
  6. Polar SPE
  7. Polar Sample Pretreatment
  8. Polar Sorbent Conditioning, Equilibration and Sample Loading
  9. Polar Sorbent Washing and Analyte Elution
  10. Cation-Exchange SPE
  11. Cation-Exchange SPE Sorbents
  12. Cation-Exchange Sample Pretreatment
  13. Sorbent Conditioning, Equilibration and Sample Loading
  14. Cation-Exchange Washing and Analyte Elution
  15. Anion-Exchange SPE
  16. Anion-Exchange SPE Sorbents
  17. Anion-Exchange Sample Pretreatment
  18. Anion-Exchange Sorbent Conditioning, Equilibration and Sample Loading
  19. Anion-Exchange Washing and Analyte Elution
  20. Minimum pKa Differential
  21. Mixed-Mode SPE
  22. Mixed-Mode SPE Sorbents
  23. Mixed-Mode Sample Pretreatment
  24. Mixed-Mode Sorbent Conditioning Equilibration and Sample Loading
  25. Mixed-Mode Washing and Analyte Elution
  26. Assessment
Method Development
  1. Learning Aims & Objectives
  2. Sorbent Column Processing
  3. Centrifugal Column Processing
  4. Vacuum Column Processing
  5. Disadvantages of Vacuum Column Processing
  6. Sorbent Column Processing
  7. Sorbent Bed Geometry
  8. Disk Products for SPE
  9. Disk Based Sorbents
  10. Fines
  11. Fines - Channelling
  12. SPE Method Development - Overview
  13. Analyte Assessment I
  14. Analyte Assessment II
  15. Analyte Assessment III
  16. Mechanism Selection I
  17. Mechanism Selection II
  18. Sorbent Screening - Alternative Sorbents I
  19. Sorbent Screening - Alternative Sorbents II
  20. Sorbent Screening Process
  21. Procedure Optimization I
  22. Procedure Optimization II
  23. Procedure Optimization III
  24. Soak Steps
  25. Drying Steps
  26. Elution Optimisation
  27. Elution Optimisation - Results
  28. SPE Method Troubleshooting Overview
  29. Recovery Problems I
  30. Recovery Problems II
  31. Recovery Problems III
  32. Reproducibility Problems I
  33. Reproducibility Problems II
  34. Cleanliness Problems
  35. Cleanliness Problems - Alternative Solvents
  36. Cleanliness Problems - Poor Sorbent Selectivity
  37. Inadequate Throughput
  38. Generic Methods in SPE - Overview
  39. Mixed Mode Sorbents
  40. Generic Methods - Overview
  41. Generic Methods - Basic Protocol
  42. Generic Methods - Further Optimization
  43. Assessment
Primary Sample Preparation Techniques
  1. Primary Sample Prep Techniques
  2. Sample Dilution in Pharmaceuticals
  3. Sample Filtration and Ultrafiltration
  4. Centrifugation and Ultracentrifugation
  5. Protein Precipitation
  6. Extraction Approaches to Sample Preparation
  7. Selectivity
  8. The Partition Coefficient
  9. Liquid / liquid Extraction
  10. Support-Assisted Liquid/Liquid Extraction
  11. Solid Phase Extraction
  12. Assessment
Liquid / Liquid Extraction Techniques
  1. Liquid / liquid Extraction II
  2. Liquid / liquid Extraction III
  3. Drawbacks of Liquid/liquid Extraction
  4. Emulsions
  5. Support-Assisted Liquid / liquid Extraction II
  6. Support-Assisted Liquid / liquid Extraction III
  7. Assessment
Approaches to Automation for SPE
  1. Automation via On-line Solid Phase Extraction
  2. Mechanisms for On-line SPE
  3. Turbulent Flow Chromatography
  4. Molecular Imprinted Polymer Sorbents (MIPS)
  5. Selecting a Sample Prep Technique
  6. Summary of Sample Prep Technique Features
  7. Automation of Sample Preparation I
  8. Automation of Sample Preparation II
  9. Assessment
Sample Content
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