Innovative Solutions - Discovery Bioanalysis

Small Polar Analytes

Rapid LC-MS/MS quantification required for a series of small polar analytes in dried blood spots (DBS). Standard generic chromatography routinely used in early screening discovery bioanalysis was incompatible due to the physicochemical properties of the analyte series.


The incompatibility of generic reverse-phase chromatography necessitated the rapid development of an LC-MS/MS method using hydrophilic interaction chromatography (HILIC). The new method employed a conventional HILIC column and resulted in good retention and chromatographic peak shape for all analytes under investigation.

However, for some of the compounds, interference from co-eluting metabolites necessitated further method development, resulting in the use of a fused core pentafluoro- phenyl (PFP) column with an isocratic mobile phase of 95% acetonitrile/5% ammonium formate (200mM). In this way, excellent retention, separation and peak shape was achieved for all analytes and all samples were turned around within four working days of compound receipt.


The use of a HILIC system, supplemented by an innovative PFP separation enabled the generation of robust, reliable quantitative LC-MS/MS data for a range of small polar analytes in DBS. Rapid method development ensured delivery of project-critical PK data in a timeframe compatible with the early discovery program.