Abstract
Silver-ion high-performance liquid chromatography (Ag-HPLC) coupled to mass spectrometry provides a unique capability to separate molecules according to the number, positions, and geometry of double bonds (DBs), which can be applied for characterization of the DB arrangement in triacylglycerols, fatty acid methyl esters, and other unsaturated molecules with low to medium polarity. Ag-HPLC is the most reliable technique for the regioisomeric separation of triacylglycerols. Ag-HPLC provides a high orthogonality to reversed-phase HPLC, therefore various configurations of two-dimensional coupling of silver ion and reversed-phase modes give a higher peak capacity for the comprehensive analysis of glycerolipids. The governing factor for the retention order in Ag-HPLC is the total number of DBs, which causes the separation of complex mixtures into distinct groups differing in the DB number. The second factor is the DB geometry, where the retention is significantly lower for transisomers and finally the smallest effect on the retention is observed for regioisomers.
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