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| Course Title: |
Practical LC-MS Method Development for Small Molecules |
| Categories: |
1 - Liquid Chromatography
2 - Liquid Chromatography-Mass Spectrometry
3 - Mass Spectrometry
4 - Sample Preparation
5 - Validation |
| Instructor(s): |
Perry Wang |
Course Number: |
6 |
| Affiliation: |
US FDA |
| Course Date: |
03/06/2016 - Sunday |
Course Length: |
1 Day Course |
| Start Time: |
08:30 AM |
End Time: |
05:00 PM |
| Fee: |
$550 ($750 after 2/12/16) |
Textbook Fee: |
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Course Description
| This course offers practical training for the practical scientists. It will take the participants step-by-step through the concepts and techniques to develop LC-MS methods. The emphasis is on practical issues associated with developing LC-MS method for small molecules. It also emphasizes problem-solving skills with examples encountered in the pharmaceutical industry and other fields. This course will provide the participants with an updated overview and a solid working knowledge of LC-MS. The participants will learn useful theoretical concepts, instrumental fundamentals and operating principles, column basics and selection guide, and key applications. After this course, the participants will be able to independently develop their LC-MS methods for small molecules. New technologies and techniques, such as monolithic chromatography and hydrophilic interaction liquid chromatography (HILIC) will be presented. Since the pharmaceutical industry is regulated by GLP and GMP, some regulation and validation concepts will be introduced. |
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Target Audience
| This one-day course is intended for analytical chemists, supervisors, lab managers, and researchers using LC-MS. It will benefit the scientists ranging from college graduates to professionals in the analytical field. |
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Course Outline
1. Introduction and Overview
History, advantages, limitations, and modes
Major modules and their function
2. Key Concepts
Retention time (tR),
Retention factor (k’)
Separation factor (α)
Column efficiency (N)
Resolution (R)
3. Mobile Phase Factors
Mobile phase selection
Organic modifiers
pH buffers
pKa/pKb of analytes
4. Operating Parameters
Flow rate
Gradient time
Column temperature (T)
5. Column Fundamentals and Selection
Packed columns (support type, dimensions and particle size and pore size)
Monolithic columns
HILIC columns
6. van Deemter Equation
Introduction of UPLC
Introduction of UHPLC
7. Mass spectrometer (MS)
Fundamental – mass resolution and mass accuracy
Triple quadropole, TOF, Ion trap and Q-exactive
How to tune and optimize an MS
8. Method Development Approaches
Gathering pertinent analyte and sample information
Finding or estimating pKa or pKb of the analytes
Defining method type (reversed phase or normal phase or HILIC)
Estimating buffer pH
Scouting gradient to get the first chromatogram
Fine-tuning and optimizing the method – solvent type and strength
9. Method Validation
Accuracy
Precision
Linearity
Weighting factors
10. Sample Preparation and Introduction of High Throughput Analysis
Protein precipitation
Liquid-liquid extraction
Solid phase extraction
Turbo-Flow technique
11. Special Topics
Monolithic chromatography
Hydrophilic interaction liquid chromatography
Core-shell technology |
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Course Instructor's Biography
| Dr. Perry G. Wang is a research chemist in the Office of Regulatory Science, the US Food and Drug Administration (US FDA). His interests include analytical method development and validation for drugs and constituents of dietary supplements and cosmetic products using GC-MS/MS and LC-MS/MS. His expertise focuses on high-throughput bioanalytical method development and validation for the pharmaceutical industry by LC-MS/MS. He received his B.S. degree in chemistry from Shandong University. He earned his M.S. and Ph.D. degrees from Oregon State University. |
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