This almost sounds like a line from Shakespeare, but HPLC or high-performance liquid chromatography and ultra-high-performance liquid chromatography (UHPLC) came a little after the ‘Bard of Avon.’ Both HPLC and UHPLC are common techniques in the analytical laboratory, but recent years have seen a shift from HPLC to UHPLC. Why is this? What are the differences between the two and how do you transfer a method from HPLC to UHPLC? These are topics I will briefly discuss in this article.
These are the top 5 differences between HPLC and UHPLC:
These are the common differences between HPLC and UHPLC, but they can also be broadly separated by application area. HPLC is commonly used in routine environments, whilst UHPLC is more common in research and development, but this is not exclusive.
As alluded to earlier in the article, there has been a gradual shift to UHPLC from HPLC over the last decade. So why is this? There are a number of reasons:
If UHPLC offers lower costs, increased speed and throughput and better results then why hasn’t everyone switched over from HPLC? Possibly because of two reasons: firstly, to achieve the higher backpressures and maintain accuracy and precision at lower flow rates then higher performance pumps and components are required which add cost to the instrument. However, even with a relatively low sample volume the higher throughput and reduced solvent costs will soon compensate for the initial higher investment in a UHPLC instrument. Secondly, as HPLC preceded UHPLC, most methods have been developed on HPLC and to transfer these over to UHPLC takes time and resources to optimise.
Transferring a method from HPLC to UHPLC is not as daunting as many fear if a few considerations are taken. Many methods can be quickly and easily transferred, and as shown above, the benefits of moving to UHPLC can be considerable. The topic of method transfer is an article in itself (and more), so here I would like to direct you to two on-demand webinars covering this topic entitled ‘From HPLC to UHPLC: How Fast Can I Be, and Is the Fastest Always the Best’ and ‘From HPLC to UHPLC: What are the Instrument Requirements and Pitfalls?’ These on-demand webinars are available in both English and German languages.
This is a question that gets asked frequently: If I have a UHPLC instrument, can I also still use it for HPLC methods? The answer is simply ‘yes’ as there should be no technical limitations to prevent this. One consideration though, in a small number of cases, is the tubing as this is likely to be narrow UHPLC tubing that could need to be exchanged for standard HPLC tubing depending on the sample solvent used. One very interesting application note caught my attention recently that showed it was possible to do both HPLC and UHPLC on the same instrument in parallel. Using the Dual LC workflow on the Vanquish Duo UHPLC System means you can have both, and at the same time.
In summary, there are many advantages to switching from HPLC to UHPLC, but frequently the barriers are the initial investment and the transfer of methods. However, the lifetime cost of UHPLC is typically lower and there are many sources of help and advice to help transfer methods from HPLC to UHPLC. Finally, a UHPLC system does offer some flexibility to perform both HPLC and UHPLC applications and with the Vanquish Duo you can run both in parallel.
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