Short Course A

Practical LC/MS: Fundamentals, Techniques and Applications

November 5 and 6

two day course
November 5-6, 2012, 8.30-12.00; 13.00-16.30
Instructor: Jack Henion

Learn effective techniques for analyzing complex samples and characterizing organic compounds using this versatile combination of analytical techniques.

Who Should Attend
Researchers, practitioners, or others involved in identifying or analyzing organic compounds in real-world samples including drugs and metabolites within or for the pharmaceutical industry, and who work with HPLC and MS techniques. Scientists who are interested in the agrochemical and industrial chemical fields will also benefit from this course, although the focus is on pharmaceutical applications and problems.
The course is designed as a practical, comprehensive introduction, which should enable the interested student to begin using the combined analytical techniques of HPLC and MS. It is desirable, but not essential, that participants have a basic knowledge of HPLC and atmospheric pressure ionization mass spectrometry.

Key Topics You’ll Learn About
Strategies for sample preparation optimized for LC/MS and LC/MS/MS analyses
How to perform on-line LC/MS and LC/MS/MS experiments for solving real-world problems
Strategies for optimal quantitation employing LC/MS and LC/MS/MS techniques
The latest developments in the area
New developments in LC/Ion Trap MS, LC-TOF/MS, high resolution MS and chip-based LC/MS techniques

How You’ll Benefit from This Course
Get solutions to your analytical problems from a leading expert
Receive a systematic review and comparison of all current commercial approaches
Learn specific LC/MS and LC/MS/MS techniques for characterizing or identifying components in complex samples
Examine specific examples from applications in the environmental, biotechnology, pharmaceutical, and chemical industries, including trace analyses
Pick up helpful suggestions and hints to help solve your own analytical problems

About the Instructor
Jack Henion is Professor Emeritus at Cornell University in the Analytical Toxicology Section of the Diagnostic Laboratory within the College of Veterinary Medicine. He is also a co-founder, Chairman and CSO of Advion BioSciences (ABS), Inc., located in Ithaca, New York. Advion is a premier contract service laboratory specializing in LC/MS/MS analyses for the pharmaceutical and agrochemical industries. Avdion has developed proprietary chip-based nanoelectrospray devices that may be used in future ‘Lab-on-a-Chip’ strategies for LC/MS analyses. Dr. Henion’s work in LC/MS relates to the analysis of real-world samples common to pharmaceutical, environmental, and biochemical problems. The instructor has published extensively in the areas of conventional capillary GC/MS as well as LC/MS, SFC/MS, IC/MS, and CE/MS using atmospheric pressure ionization (API) technologies with quadrupole, ion trap, and time-of-flight mass spectrometers.

Overview of the course

Introduction to HPLC and MS
• HPLC pumps, injectors, columns, detectors, solvents, connections, helpful hints
• MS inlet systems, mass analysis, detectors, ionization modes
• Other ionization modes and their compatibility with LC/MS 

• Principles of tandem mass spectrometry (MS/MS)

Fundamentals of LC/MS
• Nebulization principles
• Ionization mechanisms
• Generation of small droplets and aerosols
• Effects of pH, buffers, ionic strengths, and eluents
• Application problems

Atmospheric Pressure Chemical Ionization (APCI)
• Mechanism of APCI
• Basic principles
• Gas-phase chemistries
• Applications
• Problem solving

Electrospray Introduction
• Basic chemistry concepts
• Basic technical principles
• Applications
• Techniques and strategies for success
• Quantitative analysis applications
• Application problems
• Interpretation problems

Electrospray/Ion Spray of Large Molecules
• Basic principles
• Multiple charging
• Empirical determination of high molecular weights 

• Peptide and protein characterization 

Electrospray/Ion Spray of Small Molecules
• Basic principles and strategies
• Applications
• Techniques for success
• Quantitative analysis applications
• Applications problems

Qualitative LC/MS
• Determination of compound structural features
• Collision-induced dissociation (CID)—‘up-front’ CID, tandem mass spectrometry (MS/MS)
• Positive and negative ions
• The nitrogen rule
• Types of fragmentation
• Common neutral losses
• Interpretation problem solving

Sample Preparation for LC/MS
• Types of extraction
• Sample types
• 96-well format sample preparation
• Automated sample preparation—off-line, on-line
• High-throughput analysis strategies

Quantitative LC/MS Techniques
• Basic concepts
• SIM vs. SRM
• Precision and accuracy
• Calibration curves, standards, and QC samples
• Fit-for-Purpose concepts
• Applications
• Automation
• GLP issues
• Application problems

Recent Developments in LC/MS
• Nanoelectrospray techniques
• Packed capillary LC/MS techniques
• API ion trap/MS techniques and applications
• Column switching techniques for LC/MS
• Problem solving UPLC

Future Trends
• High-throughput LC/MS
• Automation techniques
• Miniaturization strategies
• Chip-based LC/MS

Revised December 2011