What HPLC Operators Need to Know Part I
Everything Needed to Understand Mobile Phase, Pumping Systems and Autosamplers
All operators of (U)HPLC instrumentation should possess significant knowledge and understanding in order to operate their instrumentation in the most efficient and effective manner. This mini-series of two webcasts will detail all critical parameters in setting up the instrument for operation, performing the analysis and ensuring the system has been left in a fully functioning capacity and ready for its next user.
Correct (U)HPLC set-up is essential, not only for the immediate analysis , but to ensure instrument down-time is reduced and up-time is increased. Good instrument operation always starts with appropriate mobile phase preparation. The majority of methods rely on the mobile phase being pH modified and controlled to facilitate acceptable peak shape and selectivity. Knowledge of how and why mobile phase pH’s are adjusted and effectively controlled is imperative to method robustness and system performance. How, when and why mobile phases are degassed and ensuring mobile phase compatibility with the prior system solvent is critical for baseline stability and peak response reproducibility. Correct mobile phase filtration is also pivotal to guard against pump wear and tear and avoid blockages.
The pump truly is the ‘heart’ of the instrument and its correct performance is vital for precise and accurate separations. Instrument operators should understand the components of the modern (U)HPLC pump and the role each element plays. The difference between isocratic, quaternary and binary pumps should be understood and specific modes by which each design function appreciated. Each design not only have their own individual benefits but also areas that may require additional set-up and optimization or maintenance. Finally, as the pump is hardest working, mechanically speaking, part of the instrument, it requires the greatest amount of maintenance. An effective preventative maintenance schedule is desired such that as much maintenance is planned prior to part component failure so that further components are alleviated and important chromatographic analysis are not compromised.
The injector introduces the sample into the mobile phase stream however the means by which this is accomplished is often overlooked by instrument operators. An understanding of all components and their individual and specific roles in this operation are beneficial as only when they’re fully appreciated can they be effectively optimized and maintained.
A good and competent (U)HPLC operator will have a sound understanding of mobile phase preparation and instrument set-up. They will also possess a good understanding of the pump and autosampler, not only they are optimized, but of any necessary maintenance, even if they are not directly involved in performing this.
Are you ‘fit for purpose’? This series of webcasts will ensure you are!
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