LC/MS Theory and Applications:

Course Description:

This 2-day course is designed to introduce the student to liquid chromatography/ mass spectrometry (LC/MS) and it's capabilities to solve real world problems related to pharmaceutical and biochemical analysis. This beginner to intermediate level lecture course will cover fundamental and practical information for using electrospray and atmospheric pressure chemical ionization LC/ MS. Practical information regarding sample preparation, adapting LC conditions for LC/MS, developing LC separations for LC/MS, the coupling various modes of chromatography (e.g. reverse, normal, size exclusion, affinity, ion pair, and ion exchange), with column diameters ranging from 0.32 to 4.6 mm, to the LC/MS interface, qualitative data interpretation, quantitation using LC/MS, and trouble shooting the LC/MS method and instrumentation. The course will have problem sets in LC/MS methods development, spectral interpretation and quantitation to emphasize learned material.

Course Outline:

Introduction Into LC/MS

Electrospray and APCI Mass Spectrometry:

Theory of Ionization
Ionization in Solution
Nebulization and Desolvation
Ion Evaporation
Gas Phase chemistry
Sample and Solution Considerations
Sample pH
Flow Rate
Electrospray Spray Chamber Hardware Configurations
Sample Introduction (orthagonal needle configuration)
Electrospray and APCI Interfaces
When Electrospray Ionization Fails
Sample Consideration
Solution Considerations
Adduct Formation

On-Line LC/MS:

Hardware Consideration for Coupling LC to MS
Mobile Phase Considerations
Flow Rates
What to Do When Incompatibilities Exist Between LC and MS
Mobile Phase compatibilities
Post Column Addition
Starting Over: Developing a New Separation

Introduction to the Interpretation of Electrospray Mass Spectra:

Generation of Structural Information by Tandem MS
Hardware Configuration
CID Energies and Transmission
Generation of Structural Information in Electrospray Transport Region
Hardware Configuration
CID Energies and Transmission
Comparison to MS/MS
Interpretation of Low Molecular Weight Environmental Compounds

LC/MS Quantitation:

Method Validation
Internal Standards
Sources or Error

Sample Preparation for LC/MS:

Techniques of Solid Phase, Liquid-Liquid, Ultrafiltration, Immunoaffinity, Column Switching, On-Column Concentration

Examples of On-Line LC/MS:

Reverse Phase Separation Using:
4.6 mm i.d. columns, 2.1 mm i.d. columns and 1.0 mm i.d. columns
Ion Pair Separations
Normal Phase Separation
Other Modes - Ion Exchange
High Throughput Analysis
Capillary Separations (CEC, Capillary Electrophoresis, and Capillary LC)

Summary/Concluding Remarks:

Future Trends
Ion Traps, time of flight MS, chip techniques
Method Development Problems to Review Materials Covered in Course

Student Profile:

  • This course is designed for the student who has completed an LC/MS short course or who has working knowledge of LC/MS and wants to improve their qualitative interpretation skills for problem solving in LC/MS.

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