High-Performance Liquid Chromatographic Determination of Eicosapentaenoic Acid in Serum by a Chemiluminescence Labeling Method

• Published in: Chemical & pharmaceutical bulletin Vol. 36; no. 5; pp. 1905 - 1908
• Main Authors: YUKI, HIDETAKA, AZUMA, YUTARO, MAEDA, NORIO, KAWASAKI, HIDEKI
• Format: Journal Article
• Language: English
• Published: Tokyo The Pharmaceutical Society of Japan 1988; Maruzen; Japan Science and Technology Agency
• Subjects: Analytical biochemistry: general aspects, technics, instrumentation; Analytical, structural and metabolic biochemistry; Biological and medical sciences; Chromatography, High Pressure Liquid; Eicosapentaenoic Acid - blood; Fatty Acids, Nonesterified - blood; Fundamental and applied biological sciences. Psychology; Humans; Luminescent Measurements; unsaturated fatty acid; Characterization; Human; Reversed phase chromatography; HPLC chromatography; Unsaturated fatty acid; Serum; Chemiluminescence; Chemical labelling; Fatty acids; Eicosapentaenoic acid
• ISSN: 0009-2363; 1347-5223
• DOI: 10.1248/cpb.36.1905

A sensitive method for the determination of eicosapentaenoic acid (EPA) in human serum and the detection of some other fatty acids was developed. Fatty acids were labeled with N-(4-aminobutyl)-N-ethyl-isoluminol (ABEI), and separated by reversed-phase high-performance liquid chromatography. The eluate was mixed with 0.2 μM microperoxidase solution and 90 mM hydrogen peroxide solution successively, and the chemiluminescence was detected in a flow cell placed in front of a photomultiplier. The calibration curve of EPA was linear in the range of 2 pmol to 2 nmol. The recovery of EPA added to human serum was nearly 100%, and the detection limit was 200 fmol. Eleven fatty acids were detected in human serum samples by this method.