Simultaneous Determination of 8, 11, 12, 20-HETEs and 13 s-HODE in Paraquat and Diquat Poisoning Patients Via the UPLC–MS/MS Method

Abstract An Ultra Performance Liquid Chromatography - Mass Spectrometry/Mass Spectrometry (UPL­MS/MS) method for simultaneous determination of 8, 11, 12, 20- hydroxyeicosatetraenoic acids (HETEs) and 13 s-hydroxyoctadecadienoic acid (13 s-HODE) in plasma was developed and optimized. The chromatographic separation of 8, 11, 12, 20-HETEs and 13 s-HODE was achieved on a Peptide Bridged Ethylene Hybrid Particles (BEH) C18 column (2.1 mm × 150 mm, 1.7 μm) with mobile phase A (Water containing 0.1% formic acid) and mobile phase B (acetonitrile containing 0.1% formic acid). HETEs and 13 s-HODE were quantified using multiple reaction monitoring mode. The developed UPLC-MS/MS method was applied to determine the plasma samples of 32 paraquat (PQ) poisoning patients, 20 diquat (DQ) poisoning patients and 38 healthy subjects. The results showed that the 8, 11, 12, 20-HETEs and 13 s-HODE have good linearity (R2 > 0.99) from 0.1–500 ng/mL. The levels of HETEs and 13 s-HODE were dramatically increased in PQ and DQ poisoning patients. There were high correlation between 8, 11, 12, 20-HETEs and 13 s-HODE. Orthogonal partial least squares discrimination analysis showed that three groups were distributed in different areas and separated well. In conclusion, a sensitive UPLC-MS/MS method had been developed and validated for determination of 8, 11, 12, 20-HETEs and 13 s-HODE in plasma of PQ and DQ patients.