Determination of four omega-3 polyunsaturated fatty acids by UPLC-MS/MS in plasma of hyperlipidemic and normolipidemic subjects

• Published in: Journal of chromatography. B, Analytical technologies in the biomedical and life sciences Vol. 1126-1127; p. 121762
• Main Authors: Zhou, Beibei, Lin, Chao, Xie, Saili, Zhou, Xingxing, Zhang, Fan, Ye, Xuemei, Lin, Feiyan, Hu, Lufeng, Huang, Aifang
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier B.V 15.09.2019
• Subjects: acetonitrile; Adult; alpha-linolenic acid; ammonia; blood lipids; blood plasma; chemical species; Chromatography, High Pressure Liquid - methods; Determination; docosahexaenoic acid; docosapentaenoic acid; eicosapentaenoic acid; Fatty Acids, Omega-3 - blood; Female; hematologic tests; Human; Humans; Hyperlipidemias - blood; ionization; kidneys; Limit of Detection; Linear Models; liver; Male; mass spectrometry; Middle Aged; monitoring; omega-3 fatty acids; Plasma; Polyunsaturated fatty acids; Reproducibility of Results; standard deviation; Tandem Mass Spectrometry - methods; ultra-performance liquid chromatography; UPLC-MS/MS; Human; Plasma; UPLC-MS/MS; Determination; Polyunsaturated fatty acids
• ISSN: 1570-0232; 1873-376X; 1873-376X
• DOI: 10.1016/j.jchromb.2019.121762

Omega-3 polyunsaturated fatty acids (PUFAs), including alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and docosapentaenoic acid (DPA), play critical roles in numerous biochemical reactions. Our aim is to develop a rapid and sensitive method for simultaneous determination of ALA, EPA, DHA and DPA in the plasma of hyperlipidemic and normolipidemic subjects. An ultra-high-performance liquid chromatography-tandem mass spectrometric (UPLC-MS/MS) method of ALA, EPA, DHA, and DPA was developed with chlorzoxazone as the internal standard (IS). The analytes were separated on an Acquity BEH C18 column (2.1 mm × 100 mm, 1.7 μm) with gradient elution by acetonitrile and 0.1% ammonia water. ALA, EPA, DHA, DPA, and IS were determined by negative electrospray ionization (ESI-) with multiple reaction monitoring (MRM) at m/z 277.42/259.05, 301.20/257.00, 327.30/283.40, 329.24/285.32, and 168.03/132.02. A total of 80 normolipidemic subjects and 83 hyperlipidemic subjects, who underwent testing for plasma lipids, liver and kidney functions, and blood routine blood test (BRT), were enrolled. There was good linearity for ALA within 1–10 μg/mL, and EPA, DHA and DPA were within 0.125–10 μg/mL. The relative standard deviation (RSD) of precision was below 15%. The concentrations of ALA, EPA, DHA and DPA were 3.47 ± 2.58, 0.41 ± 0.26, 2.93 ± 1.39 and 0.25 ± 0.21 μg/mL, respectively, in normolipidemic subjects, increasing to 4.14 ± 3.71, 0.57 ± 0.46, 3.43 ± 2.13, 0.27 ± 0.25 μg/mL, respectively in hyperlipidemic subjects. Among them, only the EPA concentration was significantly different between two groups. There was a high correlation between ALA, EPA, DHA and DPA. We developed a rapid and sensitive method for simultaneously determination of ALA, EPA, DHA and DPA in hyperlipidemic and normolipidemic subjects. In hyperlipidemic and normolipidemic subjects, concentrations of ALA were highest, followed by DHA, EPA and DPA; there were high degrees of correlation between each value. •A rapid and sensitive UPLC-MS/MS method for simultaneously determination of ALA, EPA, DHA and DPA has been established.•The validated UPLC-MS/MS method was applied successfully in plasma of hyperlipidemic and healthy subjects.•In hyperlipidemic and healthy subjects, levels of ALA were highest, followed by DHA, EPA and DPA.•There were high degrees of correlation between ALA, EPA, DHA and DPA.