Zwitterionic hydrophilic interaction solid-phase extraction and multi-dimensional mass spectrometry for shotgun...

Zwitterionic hydrophilic interaction liquid chromatography (ZIC-HILIC) material was used as solid-phase extraction sorbent for purification of phospholipids from Hypophthalmichthys nobilis. The conditions were optimized to be pH 6, flow rate 2.0mL·min(-1), loading breakthrough volume ⩽5mL, and eluting solvent 5mL. Afterwards, the extracts were analyzed by multi-dimensional mass spectrometry (MDMS) based shotgun lipidomics; 20 species of phosphatidylcholine (PC), 22 species of phosphatidylethanoamine (PE), 15 species of phosphatidylserine (PS), and 5 species of phosphatidylinositol (PI) were identified, with content 224.1, 124.1, 27.4, and 34.7μg·g(-1), respectively. The MDMS method was validated in terms of linearity (0.9963-0.9988), LOD (3.7ng·mL(-1)), LOQ (9.8ng·mL(-1)), intra-day precision (<3.64%), inter-day precision (<5.31%), and recovery (78.8-85.6%). ZIC-HILIC and MDMS shotgun lipidomics are efficient for studying phospholipids in H. nobilis.