Determination of Ceramides in Plasma by Solid Phase Extraction-Ultra High Performance Liquid Chromatography-Tandem Orbitrap High Resolution Mass Spectrometry
A method was established for the simultaneous detection of 10 ceramides in serum by solid phase extraction-ultra high performance liquid chromatography-orbitrap high resolution mass spectrometry.The sample was purified by a solid phase extraction column and separated by C30 chromatographic column after methanol extraction.The peak area was extracted by the exact mass of the parent ion for quantification,and the corresponding product ion for qualitative analysis.The standard curve was established by an isotope internal standard method.The solid phase extraction column and chromatographic column were selected,and the elution conditions of solid phase extraction,chromatographic conditions and mass spectrometry conditions were optimized.The matrix effects of different purification methods were compared.The 10 analytes showed good linearity within the specified range(r2＞0.99).The limit of detection(LOD)ranged from 0.5 ng/mL to 1.2 ng/mL while the limit of quantification(LOQ)ranged from 1.5 ng/mL to 3.5 ng/mL.The recovery ranged from 73.1％to 106.9％,and the relative standard deviation(RSD)ranged from 1.0％to 13.1％.This method has the advantages of easy operation,short operation time and high accuracy,which provides a novel technical support for the determination of ceramide and related research.
PlasmaCeramideSolid phase extractionHigh resolution mass spectrometry
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