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Eicosanoids Analysis Service

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  • Case Study

What are Eicosanoids and their functions?

Eicosanoids include prostaglandins (PG), thromboxanes (TX), leukotrienes (LT), and lipoxins (LX). Prostanoids refer to PGs and TXs. The names of prostanoids are given according to the number of carbon-carbon double bonds in the molecule. Because of the presence of two carbon-carbon double bonds, most of the biologically active prostaglandins and thromboxanes are series 2 molecules. The eicosanoids with four carbon-carbon double bonds are series 4 molecules. Prostaglandins are first shown being produced in the prostate gland. Thromboxanes are first shown being synthesized in platelets (thrombocytes). And leukotrienes are first identified from leukocytes. Therefore, they were given their respective names. The lipoxins are inflammation involved with eicosanoids, which are produced through lipoxygenase interactions. In response to ingestion of aspirin, the synthesis of lipoxins can be increased according. Therefore, lipoxins are powerful inflammation modulating eicosanoid compounds. Other eicosanoid derivatives include resolvins (Rv) and the protectins (PD).

Eicosanoids Analysis Service

Eicosanoids play an important role in the inflammatory responses of the joints, skin and eyes, on the induction of labor and on the intensity and duration of pain and fever. They also assist in the inhibition gastric acid secretion, regulation of blood pressure through vasodilation or constriction, and inhibition or activation of platelet aggregation and thrombosis. The eicosanoids of greatest biological importance to humans are a large number of molecules arachidonic acid derivatives in the reaction pathway from linoleic acid to arachidonic acid. There is also a small group of eicosanoids comes from eicosapentaenoic acid derived from α-linolenic acid. The main source of arachidonic acid is cellular stores release. Inside the cell, arachidonic acid localized primarily at the C–2 positions of membrane phospholipids and the release is triggered by the activation of PLA2.

Eicosanoids can be synthesized in all mammalian cells except erythrocytes. These molecules are rather powerful, minimum amount of eicosanoids can exert potent physiological effects. At the site of synthesis, eicosanoids exert their roles locally through receptor-mediated G-protein linked signaling pathways. There are two main pathways are associated with the biosynthesis of eicosanoids. Prostaglandins and thromboxanes are generated in the cyclic pathway, while the leukotrienes are produced in the linear pathway.

Eicosanoid Quantitative Analysis Services

Since eicosanoids play a crucial role in large number of disorders like asthma, cardiovascular disease, cancer and chronic obstructive pulmonary disease, the detection and quantification of these compounds are of great interest. Since the endogenous eicosanoids are of rather low concentration, sensitive and specific analytical methods are required for the reliable quantification of these compounds. HPLC-MS/MS has emerged as one of the main techniques used for eicosanoid quantification. Creative Proteomics has established sensitive, reliable, and accurate LC-MS/MS method for quantification of eicosanoids.

Platform

  • LC-MS/MS

Summary

  • Identification and quantification of eicosanoids by LC-MS/MS.

Technical Advantages

Platform advantages: orbitrap mass analyzer, ultra-high resolution mass spectrometry, high-quality data

Wide applicability: no species restriction, no standard product restriction

High throughput: detect dozens of lipid molecules at one time, saving samples and costs

Strong quantitative ability: the sensitivity can reach ppm level, and the linear range can reach 5-6 orders of magnitude

State of art facilities

Constantly optimized protocol and analytical software

Professional experiment design

Quick turnaround time

Sample requirement

Sample TypeMinimum Sample Amount
CellsNot less than 1x10^7 cells
TissuesNot less than 250 mg
Serum, PlasmaNot less than 300 μL
Feces and Intestinal ContentsNot less than 200 mg
Microorganisms (Bacteria, Fungi, etc.)Not less than 500 mg, not less than 10^7 CFU/mL
Other Liquid Samples"Urine not less than 1 mL, Saliva not less than 500 μL,
Amniotic Fluid, Bile, Cerebrospinal Fluid, Lymph Fluid, etc., not less than 300 μL"

Report

  • A detailed technical report will be provided at the end of the whole project, including the experiment procedure, GC-MS instrument parameters
  • Analytes are reported as uM or ug/mg (tissue), and CV's are generally<10%
  • The name of the analytes, abbreviation, formula, molecular weight and CAS# would also be included in the report.
Eicosanoids Quantified in This Service
(±)10-HDoHE(±)11,12-DHET(±)11,12-EET
(±)11-HDoHE(±)11-HETE(±)12(13)-EpOME
(±)12-HETE(±)13-HDoHE(±)13-HODE
(±)14,15-DHET(±)14,15-EET(±)16-HDoHE
(±)16-HETE(±)17-HDoHE(±)17-HDoHE
(±)17-HETE(±)18-HETE(±)20-HDoHE
(±)20-HETE(±)4-HDoHE(±)5,6-DHET
(±)5-HETE(±)7-HDoHE(±)8,9-DHET
(±)8,9-EET(±)8-HDoHE(±)9(10)-EpOME
(±)9-HETE11,12-EEQ11-dehydro TXB2
11-dehydro TXB311-deoxy PGF2α11-HEPE
12-HEPE12-keto-LTB4/12-oxo-LTB412-OxoETE
13,14-dihydro PGE113,14-dihydro PGF1α13,14-dihydro-15-keto PGA2
13,14-dihydro-15-keto PGE113,14-dihydro-15-keto PGF1α13,14-dihydro-15-keto PGF2α
13,14-dihydro-15-keto Prostaglandin D113,14-dihydro-15-keto Prostaglandin D213,14-dihydro-15-keto Prostaglandin E2
13-OxoETE13-OxoODE14 15 ,15-EEQ
14,15-DiHETE15 (S)--HPETE15/6-keto-PGF1α
15-deoxy-Δ12,14-Prostaglandin D215-deoxy-Δ12,14-Prostaglandin J215-HEPE
15-HETE15-keto PGA115-keto PGE1
15-keto PGE215-Keto-PGF2a16,17-EDP
17,18-DiHETE17,18-EEQ18-HEPE
19(R)-HETE19(R)-hydroxy prostaglandin E219(R)-hydroxy prostaglandin F2a
19,20-EDP20-carboxy LTB420-OH-LTB4
5 (S)-HpETE5(6)-EET5(S),14(R)-Lipoxin B4
5(S),6(R)-DiHETE5(S),6(R)-Lipoxin A45,6-DiHETE
5-HEPE5-OxoETE6-keto PGE1
6-keto Prostaglandin F1α6-trans leukotriene E48,9-EEQ
8-HEPE8-HETE8-iso PGE2
8-iso PGF2α8-iso Prostaglandin A18-iso Prostaglandin A2
8-iso-15-keto PGE28-iso-prostane9-HEPE
9-HETE9-HODE9-OxoODE
Arachidonic acidDocosahexaenoic AcidDocosapentaenoic Acid
Eicosapentaenoic AcidlecithinLeukotriene B4
LTB4LTC4LTD4
LTE4Maresin-1N-acetyl leukotriene E4
PGA2PGE1PGF1α
PGF2αPGF3αPGG2
PGH2PGJ2Prostaglandin B2
Prostaglandin D2Prostaglandin D3Prostaglandin E2
Prostaglandin E3Resolvin-D1Resolvin-D2
Thromboxane B2TXB3Δ12-PGD2
Δ12-PGJ2

Ordering Procedure:

Ordering Procedure

With integrated set of separation, characterization, identification and quantification systems featured with excellent robustness & reproducibility, high and ultra-sensitivity, Creative Proteomics provides reliable, rapid and cost-effective eicosanoids targeted lipidomics services.

Phospholipids and lysophosphatidic acids in relation to ulcerative colitis

Lipidomic Trajectories Characterize Delayed Mucosal Wound Healing in Quiescent Ulcerative Colitis and Identify Potential Novel Therapeutic Targets

Journal: International Journal of Biological Sciences

Published: 2022

Abstract:

In previous studies, only a handful of lipids have been revealed to play a role in human wound healing. However, due to the lack of suitable human models, research on phospholipids and class-specific fatty acids in human colonic mucosal wound healing remains underexplored. This study utilized ultra-high-performance liquid chromatography-mass spectrometry (UPLC-MS) to analyze the composition of phospholipids and class-specific fatty acids in two cohorts. Several phospholipids and class-specific fatty acids, particularly LPC, PC, LPA-PG, PI, PGD2, and PGE1, were identified in inflamed colonic tissues, representing a potential novel therapeutic approach for restoring inflammation in UC and subsequent epithelial healing, revealing new treatment targets for ulcerative colitis.

Methods: Targeted lipidomics

Ultra-high-performance liquid chromatography-mass spectrometry (UPLC-MS) was used to identify phospholipids and class-specific fatty acids. Clinical tissue samples were weighed and extracted with internal standards, BHT, and methanol. Samples were separated using a reverse-phase C18 column, detected under electrospray ionization (ESI) mode, and data were collected in multiple reaction monitoring (MRM) mode. Negative ion mode quantified some phospholipids and class-specific fatty acids, while positive ion mode quantified other types of phospholipids.

Results and Discussion:

A total of 98 phospholipids were detected and identified. Statistical analysis revealed significant differences in all detected phospholipid categories between day 0 and days 2 and 7 in the UC group, while there were no significant differences between day 0 and day 7 in the control group. A total of 33 class-specific fatty acids were detected and identified. It was observed that, compared to day 0, all groups showed significant reductions on days 2 and 7, with some being specific to UC, such as PGE1, 13,14-2OH-15keto-PGD2, PGJ2, 5(S)-HEPE, 13-HODE, and 14,15-di-HETE, while others were specific to the control group, such as 5(S)-HETE, 8-HETE, and 5(6)-EET. However, no differences were found when comparing UC and control groups at the three time points.

Eicosanoids Analysis Service

Based on the phospholipid OPLS-DA model, significant changes were observed between the UC and control groups, particularly with a higher number of affected phospholipids in the UC group (p < 0.05). On day 7, significant differences were noted between the UC and control groups, with various phospholipid classes, including phosphatidylserine (PS), phosphatidylinositol (PI), phosphatidylglycerol (PG), phosphatidylcholine (PC), phosphatidic acid (PA), and lysophosphatidic acid (LPA), all showing a significant decrease in levels within the UC group.

Eicosanoids Analysis Service

Eicosanoids Analysis Service

Based on the OPLS-DA model for classifying lysophosphatidic acids (LPAs), it was observed that the UC group had a greater number of significantly affected class-20 sulfonic acids compared to the control group (p < 0.05). However, there were no significant differences between the UC and control groups. Significant temporal variations in the levels of class-20 sulfonic acids were discovered during the injury and wound healing process, but there were no significant differences in metabolic trajectories between the UC and control groups.

Eicosanoids Analysis Service

Finally, based on the OPLS-DA model, a correlation analysis was conducted between wound scores (x-matrix) and phospholipids (y-matrix). The wound scores showed a high correlation with the phospholipid profile, especially with changes in sphingomyelins (SM) and phosphatidylethanolamines (PE), which were positively correlated with wound scores. In contrast, the levels of phosphatidylglycerol (PG) and lysophosphatidic acids (LPA) were negatively correlated with wound scores.

Eicosanoids Analysis Service

This study provides the first dynamic description of metabolic pathway disturbances during colonic mucosal wound healing in healthy individuals and quiescent UC patients. The analysis identified sustained inflammatory responses and delayed epithelial recovery during UC wound healing, along with UC-specific perturbations involving lipid mediators. Significant changes over time were observed in several phospholipids and arachidonic acid metabolites. Therapeutic interventions aimed at improving the low levels of LPC, PC, LPA, PG, PI, PGD2, and PGE1 in inflamed colonic tissues may represent a novel potential treatment approach for restoring inflammatory UC and subsequent epithelial healing. Such interventions hold promise for improving clinical practice and may contribute to the development of personalized medicines for a wide range of diseases, including UC.

* For Research Use Only. Not for use in diagnostic procedures.
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