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Virtual On-Site: LC Basics, Equipment, & Troubleshooting

Who should take this course?

If you use HPLC or UHPLC on the job and would like a better understanding of how it works, then "LC Basics, Equipment, & Troubleshooting" is the course for you. It's designed for chemists and biochemists who use LC as a regular part of their jobs, but technicians with some LC experience will also find the course valuable. No previous formal LC training is assumed, but much of the course will appeal to experienced chemists who want a firm grounding in the basics of LC.

What does it cover?

"LC Basics, Equipment, and Troubleshooting" explains chromatography in practical terms from the ground up. Here's what you'll learn:

  • The basic principles of LC
  • The five fundamental LC modes
  • The easy and logical way to adjust experimental variables to get a good separation
  • The best approach to use for various chromatographic applications
  • How to maximize column life
  • Special techniques such as gradient elution, quantitation, and sample pretreatment
  • The operating principles of each module in an LC system
  • Proven techniques for systematic problem-solving and instrument maintenance
  • The most effective, time-saving, money-saving approaches to preventing common hardware problems and method failures
. . . And much, much more

What will I get from this course ?

You will acquire a good understanding of the fundamentals of chromatography and learn simple tips and guidelines to make your chromatography work easier and more efficient. What you learn will demystify your instrument. You'll find that all of the perplexing and frustrating problems your experience have simple and logical solutions. And, better yet, you'll learn that most of these problems are preventable. The techniques you learn from this course will make you more effective at work and save you big headaches! If you've never taken a formal course or if you need a refresher, it's time to learn HPLC the right way. Students tell us that this "extremely practical course" is a must for everyone who uses HPLC.

When is the course available

The course is presented for groups of 5 - 30 people using Citrix GoToMeeting service. Virtual On-Site is six 2.5-hour sessions (typically 2 or 3 sessions per week). The usual lead time to schedule a course is approximately 4-6 weeks. Call us at (925) 297-5374 or email to to arrange timing, pricing, and content.

What topics are covered?

 The course content can be tailored to your specific needs. A typical outline looks like this:

Section 1. Isocratic Basics
  • Mechanism of chromatography
  • Retention
  • Selectivity
  • Efficiency
  • Resolution
Section 2. Gradient Basics
  • Parallels between gradient and isocratic strategies
  • Controlling gradients: gradient steepness
  • Controlling gradients: gradient range
  • Differences between gradient and isocratic strategies
  • Maintaining "equivalent" conditions
  • Gradient dwell volume issues
  • Gradient baseline issues
  • Equilibration time issues
Section 3. Columns
  • Particle size
  • The Van Deemter Equation
  • Column Geometry
Section 4. Reversed phase chromatography
  • Mechanism of reversed-phase
  • Reversed-phase solvents
  • Reversed-phase columns
  • Bonded-phase column chemistry
  • Silica purity
  • Temperature effects
Section 5. pH control; ion-pair chromatography
  • Effect of pH on reversed-phase
  • Controlling pH: buffer selection
  • Controlling peak tailing
  • Mechanism of ion-pair chromatography
  • Controlling selectivity: IP chain length and concentration
  • Problems with ion-pair chromatography

Section 6. Normal-phase, Ion-exchange, and size-exclusion chromatography

  • Normal-phase mechanism
  • Normal-phase vs. reversed-phase selectivity
  • Polar bonded-phase columns
  • Ion exchange capacity
  • Ionic strength, pH, effects
  • SEC, GFC, GPC overview
  • The SEC calibration curve
  • Column pore size selection
  • Minimizing secondary interactions
Section 7. Reservoirs and pumps
  • Degassing
  • Filtering
  • Bubbles and priming problems
  • Check valve issues
  • Seal lifetime
Section 8. Tubing and injectors
  • Tubing selection; extra-column volume
  • Teflon tubing issues
  • Stainless steel tubing issues
  • PEEK tubing issues
  • Compression fitting issues
  • Injection valve operation
  • Autosampler operation
Section 9. Column hardware
  • Inlet fittings and frits
  • Good column hygeine
Section 10. Detectors
  • UV-VIS
  • RI
  • Fluorescence
  • ELSD and CAD
  • ECD
  • LC-MS
Section 11. Quantitation problems; baseline problems
  • Integration
  • Baseline noise
  • Baseline drift
  • Artifact peaks
Section 12. Sample Preparation
  • Liquid-Solid Extraction
  • Liquid-Liquid Extraction
  • Solid Phase Extraction
  • Automation
Section 12. UHPLC
  • What's Different?
    • Particle Size
    • Extra-Column Effects
    • Gradient Dwell Volume
  • Transferring/Scaling Methods
    • Scaling for Efficiency, Flow, Pressure
    • Isocratic vs. Gradient
  • Possible Surprises
    • Gradient Distortion
    • Autosampler Delay
    • Temperature Issuses
    • Chromatographic Hygiene
Section 14. System Qualification
  • A suite of component-by-component and complete system tests to verify operating performance of your HPLC system.

Section 15. Troubleshooting Tips & Diagnostic Checklist
  • No peaks
  • Missing peaks
  • Extra peaks
  • Wrong retention time
  • Poor efficiency or peak shape
  • Poor resolution
  • Poor precision
  • Baseline noise or drift
  • Pressure problems
  • Leaks
  • Column lifetime

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