High-quality chromatography for bioanalysis of small molecule pharmaceuticals

Assessing the performance and robustness of the ExionLC 2.0 system

Ian Moore, Adrian M Taylor
SCIEX, Canada


During drug development, LC-MS/MS methods are required to deliver sufficient sensitivity and selectivity as well as high robustness such that large sample sets can be interrogated. High reproducibility is also required to meet bioanalytical requirements. Here, the performance of the SCIEX ExionLC 2.0 system on the SCIEX Triple Quad 5500+ system was investigated for linearity, precision and carryover for small molecule drug analysis.



Robust LC-MS/MS methods are essential to support pharmaceutical drug development through all phases from discovery to clinical trials. The bioanalytical scientist is constantly challenged to achieve sufficient sensitivity and selectivity in method development, while maintaining robustness such that the final method can be used across large sample sets. At all concentration levels across the calibration range and from many different sources of matrix, the method must provide high reproducibility in order to produce consistent results.

An important aspect of a successful mass spectrometry (MS) experiment in achieving selectivity, reproducibility and robustness is combining it with some form of high-quality, up-front sample separation. In the majority of cases, this is achieved using liquid chromatography (LC). In this technical note, key performance of the SCIEX ExionLC 2.0 system was investigated, specifically linearity, precision and carryover, for small molecule drug analysis in rat plasma. A SCIEX Triple Quad 5500+ system was used for detection.

Figure 1. Retention time precision. Retention time variability of the ExionLC 2.0 system for fluoxetine for over 50 injections is shown. It exhibited good retention time stability, well within a 2% RSD (dashed lines).

Key features of the ExionLC 2.0 system 

  • High-pressure, dual, serial piston pump rated to 860 bar at flow rates of 0.001 to 2 mL/min for maximum flexibility
  • Precise and stable solvent flow delivering less than 1% RSD retention time variation
  • Accurate and precise quantification results with linear coefficient of determination performance (r2) > 0.99 and precision <15% coefficient of variation for concentrations