Meet one of the developers of UHPLC, Jason Anspach, Ph.D.- Research and Development, Senior Scientist here at Phenomenex.
Ever read about advancements in science and wonder where and how it all started? When it comes to developments in Ultra High Performance Liquid Chromatography (UHPLC), Jason Anspach doesn’t have to wonder, because he was there. Towards the beginning of his career, Dr. Anspach was a part of the team that developed advanced chromatography column technology for UHPLC and capillary electrochromatography.
Jason started out in school wanting to be pre-med, as many in the industry do. However, he quickly learned that it was not the path for him and gravitated towards another passion of his–chemistry. During his time in academia he found himself asking, “why are we synthesizing this molecule? What can we use it for?” only to receive the answer, “because we can.”
This didn’t work for Jason. He wanted to develop methods, work towards creating something new, and find out how the things he was testing could be used. He wanted answers.
Dr. Anspach has since been developing and building chromatography columns for the last 20 years. Yet, he is always looking for the next thing, the next breakthrough in analytical sciences. To him, core-shell technology has been the biggest development in the last ten years in the field and will continue to be the leading advancements in chromatography.
“Miniature particles were an evolution, not revolution. The mass spectrometry industry will drive advancements in micro and nano columns as well.”
While we all wait in anticipation for the next breakthrough in chromatography, we asked Dr. Anspach some questions about a past breakthrough, UHPLC.
What are the main differences between UHPLC and HPLC?
A lot of people think that UHPLC stand for “Ultra High Pressure Liquid Chromatography” when in fact that is false. UHPLC stands for “Ultra high performance liquid chromatography”. The main difference between UHPLC and HPLC is the column efficiency, typically stated as the number of theoretical plates per meter. HPLC typically has efficiencies in the 100,000 – 150,000 plates per meter range, whereas a loose definition of UHPLC would be columns having efficiencies > 200,000 plates/meter.
What benefits are there to UHPLC?
The biggest benefit to UHPLC is time savings. You get the same resolving power in a shorter column which leads to shorter run times. While when it was first introduced people touted that you could get more efficiency per column if you continued to use the same length of column, however, the pressure limitations of even the Ultra High pressure UHPLC systems makes this rather impractical unless you are using core-shell columns.
What are some top UHPLC tips you would give to chromatographers?
1. The system matters!
Just because you have a UHPLC system does not mean that you are going to get UHPLC performance. Many UHPLC systems are not configured and plumbed to give UHPLC performance. If you can’t reproduce the chromatogram that is shipped with the column, there is a good chance your system isn’t performing up to par.
2. Ask yourself–How old is your buffer?
UHPLC columns typically have smaller interparticle spaces because of the small particles used to pack them. This makes them much less tolerant to particulates and other contaminants in the sample and mobile phase. A good rule of thumb is that buffers in the pH 3-9 range are good for a week, 92 are good for a month. Its also a good practice to flush your system with IPA weekly, especially if working at pH 6 – 8 where bacteria likes to grow.
3. Don’t forget about selectivity!
While having higher plates can help your resolution, nothing is going to help your separation more than having good selectivity.