The Application and Validation of HybridSPEPrecip

Authors: Merisa Moriarty Aoife Lee Brendan O’Connell Mary Lehane Helen Keeley Ambrose Furey

Publish Date: 2012/10/02

Volume: 75, Issue: 21-22, Pages: 1257-1269

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Abstract

Phospholipids have been shown to cause matrix effects particularly in liquid chromatography–mass spectrometry LC–MS analysis of small molecules This results in suppression of the analyte signal This study provides a versatile validated method for the analysis of serotonin in serum along with dopamine and melatonin using LC–MS/MS It utilises HybridSPEPrecipitation cartridges for the cleanup of serum samples This technology involves a simple protein precipitation step together with a fast and robust SPE method that is designed to remove phospholipids Serotonin and dopamine are major neurotransmitters in the brain which affect various functions both in the brain and in the rest of the body Melatonin plays an important role in the regulation of circadian sleep–wake cycle Good linear calibrations were obtained for the multiplex assay of analytes in serum samples 0021–3268 μmol L−1 R 2  = 09983–09993 Acceptable intra and interday repeatability was achieved for all analytes in serum Excellent limits of detection LOD and limits of quantitation LOQ were achieved with LODs of 32–235 nmol L−1 and the LOQs of 154–705 nmol L−1 for these analytes in serum The sample cleanup procedure that was developed provided efficient recovery and reproducibility while also decreasing preparation time and solvent use A sample storage protocol was established this was achieved by investigation of sample stability under different storage conditions Evaluation of matrix effects was also carried out and the influence of ion suppression on analytical results reported This cleanup protocol was then applied to the analysis of clinical serum samplesWe gratefully acknowledge funding from the Irish Research Council for Science Engineering and Technology IRCSET funding M Moriarty The Council of Directors Technological Sector ResearchStrand III 2006 Grant Scheme awarded to Dr A Furey is also acknowledged for funding the formation of the ‘Team Elucidate’ research group The Higher Education Authority Programme for Research in ThirdLevel Institutions Cycle 4 PRTLI IV National Collaboration Programme on Environment and Climate Changes Impacts and Responses is also acknowledged

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