E17: LC-MS Method Development for Small Molecule Pharmaceuticals

One-Day Course

Dr. Perry G. Wang, LC-MS Technical Expert, College Park, MD

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 molecule pharmaceuticals. It also emphasizes problem-solving skills with examples encountered in the pharmaceutical industry and academic research. 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 in the pharmaceutical industry and other field. 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.

This one-day course is intended for analysts, lab managers, regulators and researchers using LC-MS in the analytical laboratories. It will benefit the scientists ranging from college students to professionals in the pharmaceutical industry and other analytical field.

1. Introduction and Overview
     * History, advantages, limitations, and modes
     * Major modules and their function
     * Separation theories
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 to HPLC
     * Introduction to UHPLC
7.  Mass Spectrometer (MS)
     * Full scan
     * Product scan
     * Precursor scan
     * Selected ion monitoring (SIM)
     * Selected reaction monitoring (SRM)
     * Matrix effects – ion suppression and enhancement 
8.  Method Development Approaches
     * Gathering pertinent analyte (API) and sample information
     * Finding or estimating pKa or pKb of the analyte 
     * 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

Dr. Perry G. Wang is a research chemist in the Office of Regulatory Science, CFSAN, the US Food and Drug Administration (US FDA). Prior to joining the FDA, Dr. Wang worked in the pharmaceutical and medical-device industries. He received his Ph.D. degree at Oregon State University.  His expertise in the pharmaceutical field focuses on high throughput drug analysis and validation by LC-MS/MS. His current research at the FDA includes developing analytical methods for chemical constituents and contaminants in cosmetics and personal care products by GC-MS/MS and LC-MS/MS.