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The functional role of eicosanoids in metabolic-syndrome-related diseases has been extensively studied. However, their role in neuroinflammation and the development of neurodegenerative diseases is still unclear. The aim of this study was the development of a sample pretreatment protocol for the simultaneous analysis of PUFAs and eicosanoids in mouse liver and brain. Liver and brain samples of male wild-type C57BL\/6J mice (11\u2013122 mg) were used to investigate conditions for tissue rinsing, homogenization, extraction, and storage. A targeted liquid chromatography\u2013negative electrospray ionization tandem mass spectrometry method was applied to quantify 7 PUFAs and 94 eicosanoids. The final pretreatment protocol consisted of a 5-min homogenization step by sonication in 650 \u03bcL<jats:italic>n<\/jats:italic>-hexane\/2-propanol (60:40 v\/v) containing 2,6-di-<jats:italic>tert<\/jats:italic>-butyl-4-methylphenol at 50 \u03bcg\/mL. Homogenates representing 1 mg tissue were extracted in a single step with<jats:italic>n<\/jats:italic>-hexane\/2-propanol (60:40 v\/v) containing 0.1% formic acid. Autoxidation was prevented by addition of 2,6-di-<jats:italic>tert<\/jats:italic>-butyl-4-methylphenol at 50 \u03bcg\/mL and keeping the samples at 4 \u00b0C during sample preparation. Extracts were dried under nitrogen and reconstituted in liquid chromatography eluent before analysis. Recovery was determined to range from 45% to 149% for both liver and brain tissue. Within-run and between-run variability ranged between 7% and 18% for PUFAs and between 1% and 24% for eicosanoids. In liver, 7 PUFAs and 15 eicosanoids were quantified; in brain, 6 PUFAs and 21 eicosanoids had significant differences within the brain substructures. In conclusion, a robust and reproducible sample preparation protocol for the multiplexed analysis of PUFAs and eicosanoids by liquid chromatography\u2013tandem mass spectrometry in liver and discrete brain substructures was developed.<\/jats:p>","DOI":"10.1007\/s00216-019-02170-w","type":"journal-article","created":{"date-parts":[[2019,12,21]],"date-time":"2019-12-21T08:03:22Z","timestamp":1576915402000},"page":"2211-2223","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":14,"title":["Tissue pretreatment for LC\u2013MS\/MS analysis of PUFA and eicosanoid distribution in mouse brain and liver"],"prefix":"10.1007","volume":"412","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-3864-6424","authenticated-orcid":false,"given":"Madlen","family":"Reinicke","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Juliane","family":"Dorow","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karoline","family":"Bischof","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Judith","family":"Leyh","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ingo","family":"Bechmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Uta","family":"Ceglarek","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2019,12,21]]},"reference":[{"key":"2170_CR1","doi-asserted-by":"publisher","first-page":"860","DOI":"10.1038\/nature05485","volume":"444","author":"GS Hotamisligil","year":"2006","unstructured":"Hotamisligil GS. 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We performed this study in accordance with the European Commission Recommendation 2007\/526\/CE concerning the protection of laboratory animals. The animal experiments were authorized by the local ethics committee of the state of Saxony (Landesdirektion Sachsen, Leipzig, approval no. TVV 35\/12). Male wild-type C57BL\/6J mice were kept in the local animal facility under a 12 h light and dark cycle with free access to food and water. Mice were fed with a regular chow diet (V1124-300, ssniff Spezialdiaeten, Soest, Germany) for 16 weeks. Mice were killed under deep anesthesia.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Compliance with ethical standards"}},{"value":"The authors declare that they have no competing interests.","order":2,"name":"Ethics","group":{"name":"EthicsHeading","label":"Conflict of interest"}}]}}