Analysis of phospholipids in bio-oils and fats by hydrophilic interaction liquid chromatography–tandem mass spectrometry

• Scheduled MRM method for semi-quantitation of 14 phospholipid classes.
• Novel approach for HILIC separation of phospholipid classes.
• Phospholipid class selective internal standards for quantitation of the analytes.
• Partial validation of the method is presented.
• Method is successfully applied for the analysis of vegetable oils and fats.

A new, sensitive and selective liquid chromatography–electrospray ionization–tandem mass spectrometric (LC–ESI-MS/MS) method was developed for the analysis of Phospholipids (PLs) in bio-oils and fats. This analysis employs hydrophilic interaction liquid chromatography–scheduled multiple reaction monitoring (HILIC–sMRM) with a ZIC–cHILIC column. Eight PL class selective internal standards (homologs) were used for the semi-quantification of 14 PL classes for the first time. More than 400 scheduled MRMs were used for the measurement of PLs with a run time of 34 min. The method's performance was evaluated for vegetable oil, animal fat and algae oil. The averaged within-run precision and between-run precision were ≤10% for all of the PL classes that had a direct homologue as an internal standard. The method accuracy was generally within 80–120% for the tested PL analytes in all three sample matrices.