Principles of Chromatography

Instructor: Dr. Brian C. Smith – Big Sur Scientific

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Why Take This Course?

This two day (16 hour) online course teaches scientists how to use gas chromatography (GC) and high-pressure liquid chromatography (HPLC) to purify, identify, and quantify chemical species in samples. The course begins with a thorough grounding in chromatographic theory where analysts learn how to optimize their separations. We then launch into a detailed description of GC and HPLC instrumentation so attendees choose the best equipment and separation technique for their samples. The next section discusses how to calibrate chromatographs to insure quantitative accuracy. Students are then introduced to mass spectrometry (MS), perhaps the most important chromatographic detector, and see how GC-MS and LC-MS work. The course wraps up with a discussion of the applications of chromatography that is customized for each class. Current modules include the forensic and pharmaceutical applications of chromatography.

Who Should Take this Course:

Anyone who works in a chemical analysis lab, and uses or wants to use chromatography to purify, identify, and quantify the chemical species in samples.

How You Will Benefit From This Course

  1. Learn at a fundamental physical level how separations occur so you can rationally develop methods.
  2. Discover how to properly calibrate a chromatograph to optimize accuracy.
  3. See how interfacing a mass spectrometer to a chromatograph gives you a powerful technique for purifying, identifying, and quantifying the chemical species in a sample.
  4. Master the art of picking the right separation technique for your sample, saving time and money.
  5. Benefit from application modules customized to your samples.

I. Introduction to Chromatography

  • A. Terms and Definitions
  • B. The Separation Process
  • C. Elution Times and Capacity Factors
  • D. Peak Widths
  • E. Resolution
  • F. Optimizing Separations

II. Gas Chromatography (GC)

  • A. Sample Preparation
  • B. Instrumentation1.
    • Injectors
    • 2. Ovens
    • 3. Columns
    • 4. Mobile Phases
    • 5. Detectors
    • C. Method Development
    • D. GC-FTIR
    • E. Troubleshooting

III. High Pressure Liquid Chromatography (HPLC)

  • A. Sample Preparation
  • B. Instrumentation
    • 1. Injectors
    • 2. Pumps
    • 3. Columns
    • 4. Mobile phases
    • 5. Separation Types & Mechanisms
    • 6. Detectors
  • D. Method Development
  • E. Troubleshooting

IV. Calibrating Chromatographs for Quantitative Analysis

  • A. Accuracy and Precision
  • B. Standards
  • C. Calculating calibration lines
  • D. Method Validation
  • E. UV-Vis Detection: Beer’s Law

V. Introduction to Mass Spectroscopy

  • A. How a Mass Spectrometer Works
  • B. Instrumentation
  • C. Ionization mechanisms
  • D. Mass Separation schemes
  • E. Determining Molecular Weights and formulas
  • F. Determining Molecular Structures

VI. Gas Chromatography-Mass Spectrometry (GC-MS)

  • A. Instrumentation
  • B. GC-MS interfaces
  • C. Understanding GC-MS data

VII. High Pressure Liquid Chromatography-Mass Spectrometry (LC-MS)

  • A. Instrumentation
  • B. Interface Types
  • C. Understanding LC-MS Data

Application Modules Determined by Class Interest
VIII. Forensic Applications of Chromatography
IX. Pharmaceutical Applications of Chromatography