Short Course Listings
 
Short Course

Course Information
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: 3
Affiliation: USFDA
Course Date: 03/05/2017 - Sunday Course Length: 1 Day Course
Start Time: 08:30 AM End Time: 05:00 PM
Fee: $575 ($775 after 2/18/17) Textbook Fee:

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.

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.

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

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.