Since its inception UHPLC has gained popularity for enabling fast and efficient chromatography which allows the demands of modern day analyses to be met; namely increased sensitivity and higher throughput. However, moving from conventional HPLC systems to UHPLC is not without its problems and the requirement to adjust to a new way of working. Fundamental practices such as careful sample and solvent preparation and the use of appropriate fittings and tubing to reduce extra column volume and maximize peak efficiency will be discussed. We will also consider important principles such as pressure/selectivity effects, the practical implication of frictional heating, and instrument parameters such as system solvent wash out volume (as opposed to gradient dwell volume). Finally, initial conditions for method development in UHPLC and some simple, easy to use equations which can be utilized to facilitate method transfer from HPLC to UHPLC will be provided.

Topics include:

• Critical differences in UHPLC
• Making the right connections - fittings, ferrules, and tubing
• Extra column volume – you CAN make a difference
• Column selection – small particles or core shells – the debate
• Eluent preparation for UHPLC
• Selectivity and pressure – they are related
• Frictional heating - practical implications
• Practical detector considerations
• Taking the guess work out of initial method development
• Problem free method transfer – including wash out volume
• Assessing performance improvements when moving to UHPLC – just how U is your UHPLC?