Liquid chromatography with dual parallel mass spectrometry and 31P nuclear magnetic resonance spectroscopy for analysis of sphingomyelin and dihydrosphingomyelin. II. Bovine milk sphingolipids

Abstract

Liquid chromatography coupled to atmospheric pressure chemical ionization (APCI) and electrospray ionization (ESI) mass spectrometry (MS), in parallel, was used for simultaneous detection of bovine milk sphingolipids (BMS). APCI-MS mass spectra exhibited mostly ceramide-like fragment ions, [Cer-H(2)O+H](+) and [Cer-2H(2)O+H](+), which were used to identify individual molecular species of BMS according to fatty acyl chain length:degree of unsaturation and long-chain base (LCB). ESI-MS was used to confirm the molecular weights of BMS species. Both sphingomyelin (SM) and dihydrosphingomyelin (DSM) molecular species were identified, with DSM species constituting 20% of BMS. Approximately 56 to 58% of DSM species contained a d16:0 LCB, while 34 to 37% contained a d18:0 LCB. Approximately 26 to 30% of SM species contained a d16:1 LCB, while 57 to 60% contained a d18:1 LCB. BMS species contained both odd and even carbon chain lengths. The most abundant DSM species contained a d16:0 LCB with a 22:0, 23:0 or 24:0 fatty acyl chain, while the most abundant SM species contained a d18:1 LCB with a 16:0 or 23:0 fatty acyl chain. (31)P NMR spectroscopy was used to conclusively confirm that DSM is a dietary component in BMS.