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LC-MS/MS: Practical Method Development for the Pharmaceutical Industry

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Course Director

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Course Fee

$1650.00 Regular Registration

$1450.00 Early Bird Pricing (Register 30 Days in Advance)

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Course Brochure

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Course Description

This 10-hour course offers practical training for the practicing scientists. It will take the participants step-by-step through the concepts and techniques to develop LC-MS/MS methods. The emphasis is on practical issues associated with developing LC-MS/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/MS.

 

The participants will learn useful theoretical concepts, instrumental fundamentals and operating principles, column basics and selection guides, and key applications. After this course, the participants will be able to independently develop their own LC-MS/MS methods. New technologies and techniques, such as monolithic chromatography and hydrophilic interaction liquid chromatography (HILIC) will be presented. Since most of the participants are from the pharmaceutical industry, which is regulated by GLP and GMP, some regulation and validation concepts will be introduced.

Who Should Attend

This course is intended for analytical chemists, supervisors, lab managers, and researchers using LC-MS/MS in the pharmaceutical industry. It will benefit the scientists ranging from college graduates to professionals in the analytical field. The course will provide the analytical scientists with a clearer understanding and a solid working knowledge of the concepts, instrumentation, columns, applications, and best practices in LC-MS/MS method development.

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  • Learning Objectives

    At the end of this course attendees will:

    • Learn useful theoretical concepts, instrumental fundamentals and operating principles, column basics and selection guides.
    • Fully understand the master resolution equation and its application.
    • Be able to independently develop their own LC-MS/MS methods. 
    • Learn how to improve the throughput by taking advantage of LC and MS.
    • Learn how to use HILIC to deal with polar molecules, such as proteins and peptides.
    • Learn how to perform a qualitative and quantitative analysis by LC-MS/MS.

  • Agenda

    Section 1: Introduction and Overview

    • History of chromatography
    • Introduction of high-performance liquid chromatography
    • Introduction of mass spectrometry

    Section 2:  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

    Section 3: What you can Learn from the Course

    • What kind of columns should be selected?
    • How column physical and 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

    Section 4: Operating Parameters and Column Selection

    • Flow rate and gradient time
    • Column temperature (T) & packed columns (support type, dimensions, particle & pore size)
    • Monolithic and HILIC columns

    Section 5: Mass Spectrometer (MS)

    • Fundamental – charged species, mass resolution and mass accuracy
    • What kind of ionization should be selected – ESI, APPI or APCI 
    • Single and Triple quadrupole, TOF, Ion trap and Q-exactive
    • How to develop MS and MS-MS methods
    • How to perform a qualitative and quantitative analysis

    Section 6: 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

    Section 7: Special Topics

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

Registrant Information:

Each person attending a course will be asked to set up an Attendee Profile Account during the registration process. Creating an Account helps you view your order history and manage your training programs. If you are registering for others, please set up an Account in the Attendee’s name. If you are registering more than one person, you’ll need to set up a separate account for each Attendee.

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