Introduction
Being one of the main workhorses in today’s analytical research, the output of many laboratories is dominated by the amount of time that LC-MS systems are running smoothly. With a lot of determining factors, it is often quite a challenge to keep these systems up and running. Next to technical issues that can occur with the LC-MS system’s hard- and software, LC column failure is one of the most frequent causes of LC-MS system down time. Typical observations of LC column failure are an increase in column backpressure, deviating peak shapes or gradual shifts in retention time.
As an alternative to classical packed-bed LC columns, PharmaFluidics offers micromachined nano LC chip columns or micro pillar array columns (μPAC™) that distinguish themselves by several features. The inherent high permeability and low ‘on-column’ dispersion obtained by the perfect order of the separation bed makes μPAC™ based chromatography unique in its kind. The peak dispersion originating from heterogeneous flow paths in the separation bed is eliminated (no A-term contributions) and therefore components remain much more concentrated (sharp peaks) during separation [1]. The freestanding nature of the pillars also leads to much lower backpressure allowing the use of very long columns [2]. The result is a top performing nano LC column that is very robust and is much less prone to sample related column failure. To demonstrate this, a single μPAC™ column was operated under standard bottom-up proteomics conditions over a period of 6 months. Sequential injection of a HeLa cell tryptic digest, a blank and a Cytochtome C tryptic digest sample was performed to evaluate column robustness. In addition, several sample sets that are perceived as challenging have been injected to demonstrate the column’s resilience to sample related column failure.
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