Picolinyl ester fragmentation mechanism studies with application to the identification of acylcarnitine acyl groups following transesterification.

Acyl picolinyl esters provide excellent data to identify the structures of acyl groups. However, the mechanisms for the formation of fragment ions from picolinyl esters are unsettled. Proposed structures for fragment ions have focused on long-chain groups and may not accommodate results from medium- and short-chain acyl groups. Using deuterium-labeled organic acids, we have investigated the mechanisms for the formation of fragment ions. Based on these studies, we propose a new mechanism that is consistent with the experimental data. We then tested the mechanisms by analyzing selected acylcarnitines. Transesterification of acylcarnitines was performed by reaction with 3-pyridylcarbinol and potassium tert-butoxide in dichloromethane to produce acyl picolinyl esters. The picolinyl esters were separated and detected by gas chromatography/electron ionization-mass spectrometry. Each mass spectrum contained a series of peaks with m/z differences of 12, 13, or 14 u depending on the acyl group's chemical structure. The position of an unsaturated bond or branched methyl in the acyl group of acylcarnitine can be readily determined.