LC Basics, Equipment, & Troubleshooting

Custom Content

To find out more about customizing this course to meet your specific needs, give us a call at
(925) 297-5374

or email us at info@lcresources.com

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 info@lcresources.com 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
HPLC vs UHPLC

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
HILIC
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