Determination of Eicosapentaenoic, Docosahexaenoic, and Arachidonic Acids in Human Plasma by High-Performance Liquid Chromatography with Electrochemical Detection.
A high-performance liquid chromatography with electrochemical detection (HPLC-ECD) system was developed for the simultaneous determination of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and arachidonic acid (AA) in human plasma. In the present HPLC-ECD system, EPA, DHA, and AA were separated using a reverse-phase C30 column and detected based on the voltammetric reduction of 3,5-di-tert-butyl-1,2-benzoquinone (DBBQ). Chromatographic peak areas were proportional to the concentration of EPA, DHA, and AA from 0.75 μM to 0.1 mM (r > 0.998). The concentrations of EPA, DHA, and AA in plasma from healthy human subjects after overnight fasting were determined, and the ratio of EPA to AA was obtained by the present HPLC-ECD method, which required 40 μL of human plasma and a simple procedure of sample preparation using diethyl ether extraction. Moreover, changes in EPA, DHA, and AA concentrations in a human subject were monitored before and after fish oil supplement administration by the present HPLC-ECD system.
Department of Analytical Chemistry, School of Pharmacy, Tokyo University of Pharmacy and Life Sciences.
Akira
A
Kotani
Mizuki
M
Watanabe
Kazuhiro
K
Yamamoto
Fumiyo
F
Kusu
Hideki
H
Hakamata
D000328:Adult; D001095:Arachidonic Acids / Q000097:blood*; D001774:Blood Chemical Analysis / Q000379:methods*; D002851:Chromatography, High Pressure Liquid; D004281:Docosahexaenoic Acids / Q000097:blood*; D015118:Eicosapentaenoic Acid / Q000097:blood*; D004563:Electrochemistry; D005260:Female; D006801:Humans; D057230:Limit of Detection; D008297:Male; D013997:Time Factors; D055815:Young Adult
D001095:Arachidonic Acids; D004281:Docosahexaenoic Acids; D015118:Eicosapentaenoic Acid