Advanced Topics in HPLC

4-Day Course with 10 Hands-on Lab Experiments

I promise you will leave this class with the knowledge to take on any chromatography challange.

This course has a prerequisite.  You must have taken either the LC/GC Bootcamp or a previous course at Axion Labs.  Once you have a firm grasp of those HPLC fundamentals, you are ready for the advanced HPLC course which offers more breadth of LC topics…and 10 opportunities to get some hands-on experience with the new topics.  You’ll get a chance to 

  • Utilize HILIC to measure sugar content in adulterated fruit juices
  • Develop a method to simultaneously measure caffeine, artificial sweeteners and preservatives in soft drinks;
  • Convert HPLC methods to UPLC …and cut the analysis time in half!
  • Identify unknowns with the help from the 3-D capabilities of the Diode Array Dectctor
  • Analyze Hemp samples to determine cannabinoid concentrations
  • Tear apart and rebuild a modern HPLC Pump

And you’ll get to do all of this while having fun…the Axion way!

Course Curriculum

Day 1

9:00 Review of HPLC Resolution and Theory

10:45 Lab 1 – Advanced HPLC Parameters 

12 Lunch (provided by Axion)

1 PM Lab Review

1:30 Column Hardware

2:30 Lab 2 – Column Dimensions 

4:00 Lab Review

4:15 Solid Phase Micro Extraction (SPME)

5 PM End of Day

Day 2

9:00 Reversed Phase Mechanism and pH Effects (and HILIC?)

10:15 Lab 3 – pH Effects – Run pH and Caffeine standards at pH of 5.5, 4.7 and 4.0 

11:30 Quantitative Analysis

12 Lunch (provided by Axion)

1 PM Method Development and Gradients

2:00 Lab 4:  Qual/Quant of Soft Drinks

3:30 Lab Review

3:45 Lab 5 – HILIC HPLC of Sugars

5 PM End of Day

Day 3

9:00 Ion Chromatography – Dionex Sugars

9:30 Lab 6 – Dionex Sugars 

10:00 Sample Preparation

11:00 Lab 7 Amino Acids by Pre-Column Derivatization

12 Noon Lunch (provided by Axion)

1 PM Lab 8 – Method Optimization 

2:30 Troubleshooting

4:00 End of Day

Day 4

9:00 Troubleshooting HPLC

10:30 Lab 9 – Pump Maintenance 

11:30 Lab 10 – Detector and Injector Maintenance

1:00 End of Course

*Extra Discussion Topics and Demonstrations:  LC/MS, SPE Separation of Kool Aid, DAD

Have you taken the LC/GC Bootcamp?

The LC/GC Bootcamp is the best chromatography training to start with for most people.

Take the LC/GC Bootcamp first, then either (or both) of the Advanced Topics courses. If you follow that learning path, we promise you will gain technical mastery in both HPLC and GC. You will feel confident knowing you can solve almost any HPLC or GC challenge you face.


4-Day Advanced HPLC Course

Hands-On Experience With New Topics

Where: Axion Labs and Training Institute, Chicago Illinois

Instuctor: Dr. Lee N. Polite

Choose Your Date

Aug 15-18, 2023

Nov 13-16, 2023

Sign up for the wait list


What Our Students Say


Hi I just wanted to take a minute to tell you what an amazing course Axion’s LC/GC Bootcamp is. I was fortunate enough to attend Lee’s course in the summer of 2021.  I’m a trained chromatographer with a focus in mass spectrometry.  Despite having a PhD in the field with years of study behind me, I was able to garner new information to expand my knowledge base. I highly encourage anyone to attend his course if you are afforded the opportunity.

Tiffany Liden, Ph.D.
Postdoc Researcher, Adjunct Professor  Department of Chemistry and Biochemistry, The University of Texas at Arlington
{Over the course of my career, I have attended several GC and HPLC trainings. This was the first training in which I left the course with a level of understanding that I could maximize the equipment. Rather than leaving with a tip or a trick for method development, Dr. Polite provided a complete roadmap to method development from theory to application in the training. It was enjoyable to go from the art of method development to the science of method development. For anyone looking to learn GC/HPLC, increase their lab skill sets, or make the transition from experienced user to expert; I would highly recommend this course.
Joe Block
{I'm glad I had the opportunity to attend two of the courses Axion offers. Dr. Lee Polite is one of the best teachers I ever met and his knowledge over the LC subject is amazing. I definitely recommend anyone that wants to learn about chromatography to attend the courses offered by this institute.
Klleber Oliveira
Senior Scientist, Quality Control USANA Health Sciences Inc.
{The team at Axion Labs offers a level of personalized and high-quality instruction that is nearly impossible to find anywhere else. Given how challenging of a technique chromatography is, I was blown away by the ability of every instructor to break it down into lessons that were easy to understand and retain. I came into an Axion class knowing the bare minimum about chromatography and left with the tools/confidence needed to solve various chromatography problems in the lab.  
Stephen Braunewell
Associate Scientist II bluebird bio
{...was a game changer for my career. It single handedly helped me get every job I have ever had.  I was in metrology at APP pharmaceuticals. It helped me move to AD at a startup and later a third party field service tech and now owning my own service business. I am very thankful for my experience.
Michele McClain
Owner LakeShore Scientific
{Clearly explained topic with excellent detail, gave examples of applications relevant to my job application.
Cathleen Roush
Air Pollution Specialist Air Resource Board, California
{Great Instructor and staff! I enjoyed the "hands-on" part of this course. This course really broke down GC and the fundamentals.
Carla Reguen
Forensic Chemist Wilmington Police Dept. Crime Lab
{This course was extremely helpful in teaching the fundamentals of GC. The instructor presented the material in a fashion that was easily understandable.
Monika Kothari
Lab Technician Nalco Water

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