Page 12 - LifeScience Solution for Lipid and Lipidome

Analysis of Lipid Mediators in Human Blood Serum

Data Subcutaneous Tissue Mass Imaging of
Lipid mediator is the collective term for all lipids with bioactivity. In particular, it prostaglandins
refers to molecules that are released from cells and act by bonding with the cell HO COOH arachidonic acid membrane
OH
The 158 components in the standard mixture of lipid mediators were grouped according to 16 internal standard components. membrane receptors of other cells. Typical lipid mediators include O PGD 2 O COOH COOH phospholipid
Peaks can be identified accurately by applying retention time correction to the closely spaced retention times of the internal prostaglandin, leukotriene, platelet-activating factors (PAFs), endogenous HO HO COOH OH PGE 2 Analysis of
HO OH
standard peaks. Chromatograms detected from mouse brain tissue with 8-iso-PGF 2α and 8-iso-15(R)-PGF 2α identified or PGE 1 and cannabinoid, lysophosphatidic acid, and sphingosine 1-phosphoric acid. PGF 2α thromboxane Comprehensive
PGD 1 identified are shown below. As a comparison, chromatograms of corresponding standard mixtures are provided above the Lipid mediators (prostaglandin) that caused uterine contractions were first leukotrienes COOH Glycerophospholipids
isoprostanes OH O O
mouse brain chromatograms. These show almost the same differences in the retention time from the internal standard samples as discovered in sperm in the 1930s. Prostaglandin and leukotriene derived from HO O COOH OH TXA 2
COOH S NH COOH
those of the biological sample, which seems to confirm the reliability of identification. fatty acids serve the function of concentrating white blood cells at areas HO OH HN NH 2
LTC 4
O COOH
infected with a pathogen, but they can also cause fever and pain. 8-iso PGF 2α OH OH
COOH
Standard mixture Standard mixture Consequently, due to their many potential bioactivity effects, lipid mediators, DHA LTB 4 EPA
353.2>193.1(-) 353.2>235.1(-) their related compounds, and their receptors are actively being researched as a
8-iso-PGF 2α PGD 1 HO
PGF 2α OH Technology
8-iso-15(R)-PGF 2α PGE 1 target for drug discovery. OH COOH AEA COOH
11-β-PGF 2α 8-iso-PGE 1 RvD 1 OH Supercritical Fluid
HO HO H HO LXA 5 Lipid Analysis Using
PGE 2 -d4 resolvin N lipoxin
O
PGF 2α -d4 PGD 2 -d4
8.5 9.0 9.5 10.0 min 10.0 10.5 11.0 11.5 min
Brain Brain
353.2>193.1(-) PGF 2α 353.2>235.1(-) PGE 1
8-iso-PGF 2α Analysis of
PGE 2 -d4
PGD 1
8-iso-15(R)-PGF 2α2 Glycerophospholipids
PGF 2α -d4 PGD 2 -d4PG
8.5 9.0 9.5 10.0 min 10.0 10.5 11.0 11.5 min UFMS Technology
Fig. 1 MRM Chromatograms of 8-iso-PGF2α (left) and PGD 1 (right) from a Standard Sample Solution and Mouse Brain Lipid Extract Solution
The LC/MS/MS Method Package for Lipid Mediators Ver. 2
After excising brain, liver, and spleen tissue from a mouse and quickly freezing them in liquid nitrogen, each was weighed. Then includes methods for simultaneous analysis of 158 compounds LC/MS/MS Method Package Blood Serum
lipids were extracted by crushing the tissue and adding methanol and an internal standard mixture solution. The extract solutions of lipid mediators and related substances derived from the for Lipid Mediators Ver. 2 Analysis of Lipid
were diluted with formic acid and then purified and concentrated by solid phase extraction before simultaneous analysis of lipid arachidonic acid cascade. It allows all components to be Mediators in Human
mediators. The profiling results for 78 components quantitatively detected from the three types of tissue are shown in Fig. 2. The monitored with a 20-minute chromatogram. Therefore, it is
quantitation values obtained after creating calibration curves for all components are marked logarithmically on the vertical axis. possible to start analyses without the tedious tasks normally
Values on the vertical axis indicate the content per unit of tissue weight (fg/mg tissue). The PGE 2 concentration in the liver was required for simultaneous analysis, such as determining
0.10 pg/mg tissue and the PGD 2 concentration in the brain was 143 pg/mg tissue. It shows that profiling with a wide dynamic analytical conditions, optimizing MS parameters for each
range was possible in areas with low concentrations. compound, and so on. Compounds registered in the package
for simultaneous analysis include 87 components derived from of Glycolipids
Other Fatty Acid Arachidonic Acid EPA DHA arachidonic acid, 18 components derived from EPA, 16 Structural Analysis
metabolites metabolites metabolites metabolites components derived from DHA, 11 ethanolamides, and 26
200000
other compounds, such as fatty acid metabolites and
platelet-activating factors (PAFs).
20000
2000

200 Analysis of Fatty Acid
20 TXB1 PGE1 PGD1 TXB2 PGE2 PGD2 LTC4 PGA2 PGJ2 PGB2 TXB3 PGD3 AEA PAF Content of Human ES Cells Composition in Overall Lipid
12,13-DiHOME 9,10-DiHOME 9-HOTrE 13-HOTrE 13-HODE 9-HODE 9-HpODE 13-KODE 13-HpODE 9-KODE 15-HEDE 15-KEDE 1a1b-dihomo-PGF2 15-HETrE 6-keto-PGF1 8-iso-15(R)-PGF2 8-iso-PGF2 5-iPF2a-VI 8-iso-15-keto-PGF2 PGF2 8-iso-PGE2 15-keto-PGF2 5S,14R-LXB4 15-keto-PGE2 5S,6R-LXA4 13,14-dihydro-15-keto-PGE2 13,14-dihydro-15-keto-PGD2 8,15-DiHETE 5,15-DiHETE 14,15-DHET 12-HHT 11,12-DHET 19-HETE 15-deoxy-delta-12,14-

Brain 6,15-diketo-13,14-dihydro-PGF1 13,14-dihydro-15-keto-PGF2
Liver
LCMS-8060
Spleen LCMS-8030 LCMS-8040 LCMS-8050 LCMS 8060
Fig. 2 Lipid Mediator Profiling in Mouse Brain, Liver, and Spleen Esters by GC Acid Methyl Analysis of Fatty

Reference: Technical Report “Development of a Comprehensive Detection Method of Eicosanoids and Platelet Activating Factor Using Ultra-High Performance
Liquid Chromatography/Mass Spectrometry” (C146-E219), LC/MS/MS Method Package for Lipid Mediators Ver. 2 flyer (C146-E225) Reference: LCMS-8030 brochure (C146-E141), LCMS-8040 brochure (C146-E188), LCMS-8050 brochure (C146-E232), LCMS-8060 brochure (C146-E286)

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