In Person

Practical LC-MS Method Development for Small Molecules

About the Course

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 methods 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. Sample preparation and new techniques, such as monolithic chromatography and hydrophilic interaction liquid chromatography (HILIC) will be presented.

Recommended textbook: M. W. Dong, HPLC and UHPLC for Practicing Scientists, 2nd Ed., Wiley, Hoboken, NJ, 2019

What You Will Learn

The participants will learn:

  • useful theoretical concepts, instrumental fundamentals
  • operating principles
  • column basics and selection guides
  • key applications
  • how to independently develop LC-MS methods

Who Should Attend

Analytical chemists, supervisors, lab managers, and researchers using LC-MS. It will benefit the scientists ranging from college graduates to professional in the analytical field. 

 

Course Outline

  • Key Concepts

    • Retention time (tR),
    • Retention factor (k’)
    • Separation factor (α)
    • Column efficiency (N)
    • Chromatographic resolution (R)
    • pKa/pKb of analytes
    • van Deemter Equation
    • Fundamentals of mass spectrometry
    • Atmospheric pressure ionization (API) in mass spectrometry
    • Common ionization modes: ESI, APCI and APPI
    • Mass analyzers: quadrupole, time of flight, ion trap and orbitrap
    • Mass resolution and mass accuracy
    • Matrix effects
  • Column and Solvent Selection

    • What kind of columns should be selected?
    • How column physical property affects the resolution
    • How column chemical property affects the resolution
    • How pH affects the separation
    • How to transfer HPLC methods to UHPLC/UPLC methods
    • Which mode should be selected – isocratic or gradient
    • How to select the best solvents for LC- MS
    • How to optimize a gradient profile
    • Separation mechanism: reversed-phase or HILIC or normal-phase
    • Mobile phase selection and organic modifiers
    • How pKa/pKb affect separation
    • How to eliminate and compensate matrix effects of MS
    • Validation consideration
  • Operating Parameters

    • Flow rate
    • Gradient time
    • Column temperature (T)
    • Packed columns (support type, dimensions, particle size and pore size)
    • Monolithic columns
    •  HILIC columns
  • Method Development Approaches

    • 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)
  • Method Validation

    • Accuracy
    • Precision
    • Linearity
    • Weighting factors
  • Sample Preparation and Introduction of High Throughput Analysis

    • Protein precipitation
    • Liquid-liquid extraction
    • Solid phase extraction
    • Turbo-Flow technique
  • Special Topics

    • Monolithic chromatography
    • Hydrophilic interaction liquid chromatography
    • Core-shell technology

Dates, Locations, and Prices

Five for four! Register five people for one course, one person for five courses, or any combination in between and your fifth registration is free. The free registration will be the course of the lowest price. Please note: This discount cannot be combined with any other discount offered. 

About the Instructor(s)

Perry G. Wang

Chemist, FDA

Dr. Perry G. Wang worked in the pharmaceutical and medical-device industry from 2000 to 2008. He received his Ph.D. at Oregon State University. Dr. Wang currently works at FDA as a chemist. His expertise in the pharmaceutical field focuses on high throughput drug analysis and validation by LC-MS.