Identification of molecular species of monoalkyldiacylglycerol from the squid Berryteuthis magister using liquid chromatography-APCI high-resolution mass spectrometry.

Monoalkyldiacylglycerol (MADAG) is an important lipid class in mollusks, corals, starfishes, and some species of zooplankton. Up to 80% of liver oils of squids and sharks are comprised of MADAG. Except for one fish species, there are no data on the composition of MADAG molecular species of marine organisms. The molecular species of MADAG obtained from digestive glands of the deep-sea squid Berryteuthis magister were identified. High-performance liquid chromatography (HPLC) and tandem high-resolution mass spectrometry (HRMS) with atmospheric pressure chemical ionization (APCI) were applied. The mass spectra of each molecular species contained positive quasi-molecular ion [M+H] with fragment ions [M-R'CO], [M-R''CO], and [M-OR'''], as well as negative cluster ion [M+2HOH] without fragments. The exact masses of the negative ions allowed determination of the molecular formulas and the number of double bonds; the positive fragment ions were used to determine the location of acyl and alkyl groups in MADAG molecules. Ninety molecular species of MADAG were revealed. The regiospecific composition was identified for 72 individual molecular species. Eleven pairs of positional isomers of acyl groups were found. Four molecular ions were produced by a mixture of acyl/alkyl isomers. Most of molecular species contained 16:0e or 18:1e alkyl groups in the sn-1 position, whereas polyunsaturated fatty acids mainly esterified the sn-2 position of MADAG from B. magister. These structural features are supposed to be characteristic for biologically active MADAG.