Zwitterionic hydrophilic interaction solid-phase extraction and multi-dimensional mass spectrometry for shotgun lipidomic study of Hypophthalmichthys nobilis

•Phospholipids were purified using a ZIC-HILIC SPE cartridge.•Molecular species of each phospholipid are studied by shotgun lipidomics.•Phospholipids with polyunsaturated acyl chains were detected in H. nobilis.•The method was validated and shown to be reliable and efficient.

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 2.0 mL·min−1, loading breakthrough volume ⩽ 5 mL, and eluting solvent 5 mL. 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 34.7 μg·g−1, respectively. The MDMS method was validated in terms of linearity (0.9963–0.9988), LOD (3.7 ng·mL−1), LOQ (9.8 ng·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.