A sensitive UHPLC-QqQ-MS/MS method for the simultaneous analysis of MDA/4-HHE/4-HNE in edible oils and nuts was developed by derivatization optimization and SPE purification

A simultaneous detection of MDA/4-HHE/4-HNE method is developed by UHPLC-QqQ-MS/MS. • Optimized derivatization and purification reduced interference from lipid impurities. • This validated method showed high sensitivity/precision with low LOD/LOQ. • Micrograms per kilogram level sensitivity capably monitors lipid peroxidation in oily food. A novel and sensitive method using ultra-high performance liquid chromatography-tandem triple quadrupole mass spectrometry (UHPLC-QqQ-MS/MS) was developed for the simultaneous determination of three lipid oxidation markers: malondialdehyde (MDA), 4-hydroxy-2-hexenal (4-HHE), and 4-hydroxy-2-nonenal (4-HNE) in edible oils and nuts. Optimization of 2,4-dinitrophenylhydrazine (DNPH) derivatization and solid-phase extraction (SPE), combined with isotope compensation to ensure high analytical performance: linearity ( R  ≥ 0.9990), precision (RSD ≤ 18.75%), accuracy (SD ≤ 19.24%), recovery (80.34–119.78%). Limit of detection (LOD) and limit of quantification (LOQ) ranged from 0.25 to 0.50 μg/kg and 0.50–2.50 μg/kg, respectively. This validated method was further applied to monitor lipid peroxidation levels, revealed that thermal processing significantly increases MDA/4-HHE/4-HNE contents, which reached 7.80 mg/kg from 26.50 μg/kg in oils and 0.98 mg/kg from 8.27 μg/kg in nuts. Notably, the accumulation of MDA and 4-HHE in edible oils was approximately tenfold higher than in nuts. A novel and sensitive method using UHPLC-QqQ-MS/MS was developed for the simultaneous determination of three lipid oxidation markers: MDA, 4-HHE and 4-HNE in edible oils and nuts. Optimization of DNPH derivatization and SPE, combined with isotope compensation to ensure high analytical performance. Download: Download high-res image (119KB) Download: Download full-size image