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SIMULTANEOUS DETERMINATION OF DOCOSAHEXAENOIC ACID AND EICOSAPENTAENOIC ACID BY LC-ESI-MS/MS FROM HUMAN PLASMA

A sensitive and rapid method based on liquid chromatography/tandem mass spectrometry (LC/MS/MS) with simple single step protein precipitation has been developed and validated for the quantitative determination of docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA) in human plasma. After addition of internal standard to human plasma, samples were extracted by simple protein precipitation using acetonitrile as the precipitating agent. The extracts were analyzed by HPLC with the detection of the analyte in the multiple reaction monitoring (MRM) mode. This method for the simultaneous determination of DHA and EPA is accurate and reproducible, with limits of quantitation of 50.00 ng/mL in plasma. The standard calibration curves for both DHA and EPA are linear (r > 0.99) over the concentration ranges 50.00–7498.50 ng/mL in human plasma, respectively. The intra- and inter-day precision over the concentration range for DHA and EPA are less than 10.16 and 6.72 (relative standard deviation, %RSD), and accuracy is between 91.17–104.74% and 95.81–108.33%, respectively.