Novel characterisation of minor α-linolenic acid isomers in linseed oil by gas chromatography and covalent adduct chemical ionisation tandem mass spectrometry

•We identified 4 minor isomers of α-linolenic acid (ALA) in linseed oil.•Isomers were resolved by GC using a new and highly polar phase column (SLB-IL111).•The positional 9-12-15 C18:3 configuration was achieved by CACI-MS/MS.•Geometrical configuration was established with standards based on relative retention.•We postulated preferred and unfavoured isomerisation pathways of ALA.

Discrimination between polyunsaturated fatty acid isomers with three double bonds is a great challenge, due to structural similarities and similar polarities. In this study, we report the identification of four minor geometrical isomers of α-linolenic acid (ALA) present in linseed oil samples: (9E,12Z,15E)-, (9Z,12Z,15E)-, (9Z,12E,15Z)- and (9E,12Z,15Z)-octadeca-9,12,15-trienoic acids, chromatographically resolved by gas chromatography (GC) using a new and highly polar ionic phase column (SLB-IL111). Gas chromatography–electron ionisation mass spectrometry (GC–EIMS) determined that the four unknown compounds were C18:3 n–3 isomers. The positional 9-12-15 C18:3 configuration was achieved by covalent adduct chemical ionisation tandem mass spectrometry (CACI-MS/MS) while geometrical configuration was established with analytical standards based on relative retention. We hypothesised that these isomers are formed during linseed oil deodorisation and postulate preferred and unfavoured isomerisation pathways of ALA.