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.