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. 2023 Jun 13;28(12):4728.
doi: 10.3390/molecules28124728.

Validation of a Liquid-Liquid Extraction Method to Study the Temporal Production of D-Series Resolvins by Head Kidney Cells from Atlantic Salmon (Salmon salar) Exposed to Docosahexaenoic Acid

Pedro Araujo  1 , Sarah Iqbal  1   2 , Aleksander Arnø  1   3 , Marit Espe  1 , Elisabeth Holen  1
Affiliations

Validation of a Liquid-Liquid Extraction Method to Study the Temporal Production of D-Series Resolvins by Head Kidney Cells from Atlantic Salmon (Salmon salar) Exposed to Docosahexaenoic Acid

Pedro Araujo et al. Molecules. .

Abstract

A simple and rapid method for the extraction of D-series resolvins (RvD1, RvD2, RvD3, RvD4, RvD5) released into Leibovitz's L-15 complete medium by head kidney cells from Atlantic salmon and the further determination of liquid chromatography triple quadrupole mass spectrometry is proposed. A three-level factorial design was proposed to select the optimal concentrations of internal standards that were used in the evaluation of the performance parameters, such as linear range (0.1-50 ng mL-1), limits of detection and quantification (0.05 and 0.1 ng mL-1, respectively), and recovery values ranging from 96.9 to 99.8%. The optimized method was used to determine the stimulated production of resolvins by head kidney cells exposed to docosahexaenoic acid, and the results indicated that it is possible that the production was controlled by circadian responses.

Keywords: Atlantic salmon; head kidney cells; liquid chromatography quadrupole mass spectrometry; liquid–liquid extraction; resolvins.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Extracted ion chromatograms, in increasing order of retention times, to indicate the selectivity of the analysis towards the five analyzed resolvins, after implementing the proposed LLE protocol.
Figure 2
Figure 2
Principal component analysis of the released resolvins in L-15 media by head kidney cells with and without exposure to DHA, after implementing the proposed LLE protocol.
Figure 3
Figure 3
Concentration/time ratio for the different resolvins in the control and DHA group. The former group shows a continuous decrease in production and the latter a plausible regulated production by a circadian clock.
See this image and copyright information in PMC

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