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. 2021 Jan 11;11:410. doi: 10.1038/s41598-020-79716-x

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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LC–MS/MS analysis of 4-OXO-DHA in rat plasma. ESI negative, transition m/e = 341 → 135. Panel A represents the synthetic standard (5 ng in ethanol), while Panel B represents the detection of 4-OXO-DHA in un-spiked plasma sample from rats that received DHA. In Panel A, 4-OXO-DHA (1 mg) was dissolved in 1 ml ethanol and farther diluted to yield 1 µg/1 ml solution; 5 µl was analyzed. In Panel B, plasma sample (500–700 µl) from each rat (n = 5) mixed with 500 µl of buffer (1 M sodium acetate, pH 6) and then was extracted 3 times with 2 ml of a mixture consisted with ethyl acetate:hexane:acetic acid (75:24:1). The extract was dried over sodium sulfate, filtered and then evaporated to dryness. The residue was reconstituted in 150 µl of ethanol and 5 µl was analyzed. CPS = counts per second.