Prostaglandin F2-alpha (PGF2-α); 6-keto-prostaglandin F1-alpha (6-keto-PGF1-α); PGA1, PGB2, PGD2, PGE2, 8-isoPGF2-α, 15-deoxy-PGJ2; thromboxane B2 (TXB2); leukotriene B4 (LTB4); leukotriene E4 (LTE4); 5-hydroxyeicosateraenoic acid (5-HETE); 5,6-dihydroxyeicosatrienoic acid (5,6-DHET); 12-HETE; 8-HETE; 9-HETE; 5-oxoeicosatetraenoic acid (5-OxoETE); 15-HETE; 16-HETE; 20-HETE; 11,12-DHET, 14,15-DHET; 8,9-epoxy-5Z,8Z,11Z-eicosatrienoic acid (8,9-EET); 11,12-EET; 14,15-EET; 5-hydroxyeicosapentaenoic acid (5-HEPE); 15-HEPE; 9,10-dihydroxy-9Z-octadecenoic acid (9,10-DiHOME); 12,13-DiHOME; 9,10-epoxyoctadecenoic acid (9,10-EpOME); 12,13-EpOME; 9-hdroxyoctadecadienoic acid (9-HODE); 13-HODE; 1-cyclohexyl-dodecanoic acid urea (CUDA); arachidonic acid (ARA); PGF2-α-d4; TXB2-d4; 12,13-EpOME-d4; AA-d8; 9-HODE-d4; and linoleic acid (LA), were obtained from Cayman Chemical (Waterloo, Australia). Formic acid, hexane, acetonitrile and methanol of high-performance liquid chromatography (HPLC) grade were purchased from Sigma Co. (Santa Cruz, CA, USA).
12 13 epome d4
Manufactured by Cayman Chemical
Sourced in United States
12,13-EpOME-d4 is a deuterium-labeled fatty acid derivative. It is used as an internal standard and reference compound in analytical methods.
Automatically generated - may contain errors
Lab products found in correlation
Arachidonic acid, by Cayman Chemical (1 mentions)
Linoleic acid, by Cayman Chemical (1 mentions)
8 iso pgf2α, by Cayman Chemical (1 mentions)
Txb2 d4, by Cayman Chemical (1 mentions)
Pgf2α d4, by Cayman Chemical (1 mentions)
9 hode d4, by Cayman Chemical (1 mentions)
13 hode, by Cayman Chemical (1 mentions)
Aa d8, by Cayman Chemical (1 mentions)
Methanol, by Merck Group (1 mentions)
Formic acid, by Merck Group (1 mentions)
Hexane, by Merck Group (1 mentions)
Acetonitrile, by Merck Group (1 mentions)
12 13 dihome d4, by Cayman Chemical (1 mentions)
Oasis hlb column, by Waters Corporation (1 mentions)
Xbridge c18 column, by Waters Corporation (1 mentions)
Nexera lc 30ad, by Shimadzu (1 mentions)
Lcms 8040, by Shimadzu (1 mentions)
Labsolutions software, by Shimadzu (1 mentions)
Deuterated standard, by Cayman Chemical (1 mentions)
Acetonitrile solvents, by Merck Group (1 mentions)
3 protocols using 12 13 epome d4
1
Quantification of Lipid Mediators
2
Quantitative Analysis of Oxidized Fatty Acids
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Total lipid fractions were extracted from purified beef tallow samples by single-step deproteinization using methanol. The oxidized fatty acid fraction was purified from the lipid fractions by solid-phase extraction with Oasis HLB columns (Waters Corporation, MA, USA). Hydroxy fatty acids were separated using a high-performance liquid chromatography system (Nexera LC-30AD, Shimadzu Corporation, Kyoto, Japan) equipped with an XBridge C18 column (particle size 3.5 µm, length 150 mm, inner diameter 1.0 mm; Waters) and analyzed on a triple quadrupole mass spectrometer (LC-MS-8040; Shimadzu, Kyoto, Japan).
Mass spectrometric analysis of hydroxy fatty acids was performed in negative ion mode with an injection volume of 15 µL containing 0.5 mg of the oxidized fatty acid fraction and 1500 pg of the internal standard (12,13-diHOME-d4, 13S-HODE-d4, 13-KODE-d3, 12,13-EpOME-d4; Cayman chemicals, Ann Arbor, Michigan USA). The quantification of hydroxy fatty acids was identified and quantified by multiple-reaction monitoring as reported for the determination of other lipid metabolites [47 (link)]. For quantitation, calibration curves were prepared for each compound, and recoveries were monitored using deuterated internal standards. Data analysis was performed using LabSolutions software (Shimadzu, Kyoto, Japan).
Mass spectrometric analysis of hydroxy fatty acids was performed in negative ion mode with an injection volume of 15 µL containing 0.5 mg of the oxidized fatty acid fraction and 1500 pg of the internal standard (12,13-diHOME-d4, 13S-HODE-d4, 13-KODE-d3, 12,13-EpOME-d4; Cayman chemicals, Ann Arbor, Michigan USA). The quantification of hydroxy fatty acids was identified and quantified by multiple-reaction monitoring as reported for the determination of other lipid metabolites [47 (link)]. For quantitation, calibration curves were prepared for each compound, and recoveries were monitored using deuterated internal standards. Data analysis was performed using LabSolutions software (Shimadzu, Kyoto, Japan).
3
Oxylipidomic Measurements Utilization
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Chemicals used for this study were of following origin: Arachidonic acid (AA) and standards of 15-HETE, 12-HETE, 5-HETE, 12-HEPE, 15-HEPE, 14-HDHA, 17-HDHA, 13-HODE, 13-HOTrE) from Cayman Chem.; sodium borohydride from Life Technologies, Inc. (Eggenstein, Germany); restriction enzymes from ThermoFisher (Schwerte, Germany); isopropyl-β-thiogalactopyranoside (IPTG) from Carl Roth GmbH (Karlsruhe, Germany); E. coli (strain Rosetta2 DE3 pLysS) from Novagen (Merck-Millipore, Darmstadt, Germany). Oligonucleotide synthesis was carried out at BioTez Berlin Buch GmbH (Berlin, Germany). Nucleic acid sequencing was performed at Eurofins MWG Operon (Ebersberg, Germany). HPLC grade solvents and water were purchased from Fisher Scientific (New Hampshire, USA). The origin of other chemicals employed in this study is specified in the description of the methods. The following chemicals were used for oxylipidomic measurements: Deuterated standards (LTB4-d4, 20-HETE-d6, 15-HETE-d8, 13-HODE-d4, 14,15-DHET-d11, 9,10-DiHOME-d4, 12,13-EpOME-d4, 8,9-EET-d11, PGE2-d4; 10 ng/ml each) from Cayman Chem. (Ann Arbor, USA); acetonitrile, solvents from Merck (Darmstadt, Germany) and Fisher Scientific (Schwerte, Germany).
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