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KMID : 0370220210650060466
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Yakhak Hoeji
2021 Volume.65 No. 6 p.466 ~ p.475
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Analysis and Clinical Application of Polyunsaturated Fatty Acids in Human Serum Using One-step Extraction and Ultra-High-Performance Liquid Chromatography-Tandem Mass Spectrometry
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Lee Hye-Jin
Yoon Hye-Ran
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Abstract
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Omega-3 and omega-6 fatty acids are reported to alleviate various inflammatory reactions such as asthma,chronic obstructive pulmonary disease, rheumatoid arthritis, and inflammatory bowel disease. We have developed a newmethod for their quantitation in serum, utilizing ultra-high-performance liquid chromatography-electrospray ionizationtandemmass spectrometry (UHPLC-ESI-MS/MS) in multiple reaction monitoring (MRM) mode, under negative ionization.
MRM transitions were detected for five omega fatty acids, as follows: α-linolenic acid, m/z=277.20→277.20; eicosapentaenoicacid, m/z=301.10→257.30; docosahexaenoic acid, m/z=327.00→283.15; arachidonic acid, m/z=303.20→259.25; docosapentaenoicacid, m/z=329.10→285.25. After comparing several different solvents, we selected butanol:methanol (1:1, v/v) as theoptimal extraction solvent. Chromatographic separations were carried out using an Acclaim RSLC 120 C18 column(150×2.1 mm, 2.2 μm) at 40oC. For the mobile phase, solvent A consisted of water containing 5% acetonitrile and 5%methanol, and solvent B consisted of methyl tert-butyl ether:acetonitrile:methanol (10:20:70, v/v/v) containing 2.5 mMammonium acetate. The calibration curve showed excellent linearity within the range of 0.01-20 μg/mL (R2=0.9996-0.9999). The analytical method was validated and shown to have excellent sensitivity (LOD; 0.001-0.005 μg/mL, LOQ;0.01-0.2 μg/mL), making it suitable for targeted analyses of omega fatty acids. Recovery ranged from 78.0 to 103.9%(RSD 0.5-5.6%). This new method uses relatively small amounts of serum and extraction solvent and is expected to besuitable for analysis of various clinical specimens, such as plasma, urine, and cerebrospinal fluid.
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KEYWORD
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Omega fatty acid, One-step extraction, Negative ionization, UHPLC-MS/MS
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