| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
| Targets |
Endogenous Metabolite; The biological actions of (±)8-HETE are primarily mediated through interactions of its constituent enantiomers 8(R)-HETE and 8(S)-HETE with various signaling pathways. 8(S)-HETE has been shown to act as a selective activator of peroxisome proliferator-activated receptor alpha (PPARα) at concentrations as low as 0.3 μM and also activates protein kinase C (PKC) with an IC₅₀ of 100 μM in mouse keratinocytes. Furthermore, HETE family members play important roles in regulating transmembrane ion transport and protein kinase activity, which are crucial for cell signaling.
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| ln Vitro |
8-HETE is a known human metabolite of arachidonic acid. Plasma and urine samples were collected after four weeks and then eleven potential biomarkers (15-HETE, 12-HETE, TXA2, 5-HETE, AA, PGI2, PGF2α, 8-HETE, PGD2, PGE2 and LTB4) were identified and quantified by UHPLC-MS/MS. The chromatographic separation was carried out with gradient elution using a mobile phase comprised of 0.05% formic acid aqueous solution (pH=3.3) (A) and acetonitrile: methanol (80:20, V/V) (B), and each AA metabolites was measured using electrospray ionization source with negative mode and multiple reaction monitoring. The eleven biomarkers in BPH group rat plasma and urine were significant higher than those in sham group rats. Using the potential biomarkers as a screening index, the results suggest that ZSP can potentially reverse the process of BPH by partially regulating AA metabolism through refrain the expression of cyclooxygenase (COX) and lipoxygenase (LOX). This study demonstrates that a metabolomic strategy is useful for identifying potential BPH biomarkers and investigating the underlying mechanisms of a TCM in BPH treatment[1].
In vitro studies demonstrate that 8-HETE plays an important role in modulating human neutrophil function. In chemotaxis assays, 8-HETE (as an 85:15 mixture with 9-HETE) achieves peak chemotactic responses at 5 μg/mL, with a rank order of chemotactic potency of 5-HETE > 8-HETE:9-HETE > 11-HETE = 12-L-HETE. At optimal chemotactic concentrations, 8-HETE does not stimulate superoxide generation, alter IgG-Fc receptor expression, or evoke lysosomal enzyme release, indicating functional selectivity in neutrophil activation. Methyl esterification of 8-HETE significantly reduces its chemotactic activity (to less than 12% of the parent compound) and competitively inhibits the chemotactic activity of the free acid. |
| ln Vivo |
Zishen pill (ZSP) is a traditional Chinese medicine (TCM) used to treat benign prostatic hyperplasia (BPH). The study used a metabolomic approach based on UHPLC-MS/MS to profile arachidonic acid (AA) metabolic changes and to investigate the interventional mechanisms of ZSP in testosterone- induced BPH rats. In order to explore the potential therapeutic effect of ZSP, rat models were constructed and orally administrated with ZSP. Plasma and urine samples were collected after four weeks and then eleven potential biomarkers (15-HETE, 12-HETE, TXA2, 5-HETE, AA, PGI2, PGF2α, 8-HETE, PGD2, PGE2 and LTB4) were identified and quantified by UHPLC-MS/MS.[1]
In vivo studies demonstrate that 8-HETE production is closely associated with skin inflammation and tumor promotion processes. Following a single application of the complete tumor promoter TPA (12-O-tetradecanoylphorbol-13-acetate) to mouse back skin, 8-HETE was detected as a major arachidonic acid metabolite in epidermal extracts. 8-HETE production begins after a lag phase of approximately 3 hours following TPA treatment and reaches a maximum at 24-48 hours. Non-promoting phorbol esters are approximately 10-fold less efficient at inducing in vivo lipoxygenase responsible for 8-HETE production, while the Ca²⁺ ionophore A23187 is 100-fold less efficient. Three-day-old neonatal mice cannot be induced to generate 8-HETE in response to TPA, whereas 8-day-old mice show an extremely strong response, suggesting developmental stage-dependent regulation of this metabolite. In a rat model of benign prostatic hyperplasia, plasma and urine levels of 8-HETE were significantly elevated in the BPH group compared to sham-operated rats. |
| Enzyme Assay |
Cell-free assays for (±)8-HETE primarily employ chromatographic and spectroscopic analytical methods. A standard protocol includes: 1) Stereochemical assignment by chiral HPLC with UV detection (235 nm), utilizing hydroxypropyl-γ-cyclodextrin-modified micellar electrokinetic chromatography for simultaneous separation of (S)- and (R)-enantiomers of 8-HETE; 2) UV spectrophotometric detection at 237 nm wavelength, corresponding to the characteristic absorption maximum of its conjugated diene chromophore; 3) Quantitative analysis by LC-MS/MS using negative ion mode with multiple reaction monitoring, calibrated with deuterated internal standards such as d8-8-HETE; 4) For receptor binding studies, surface plasmon resonance or fluorescence polarization techniques can be used to evaluate interactions with nuclear receptors such as PPARα.
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| Cell Assay |
The in vitro cell assay protocol for (±)8-HETE is as follows: 1) Isolate and resuspend target cells (such as human neutrophils, keratinocytes, or macrophages) in appropriate buffer (e.g., HBSS or RPMI 1640); 2) Treat cells with various concentrations of (±)8-HETE (typically ranging from 0.1-10 μg/mL); 3) For chemotaxis assays, use Boyden chambers or Transwell systems (5 μm pore size), place cells in the upper chamber with medium containing gradient concentrations of (±)8-HETE in the lower chamber, incubate for 30-120 minutes at 37°C, and count migrated cells; 4) For chemokinesis assays, assess random migration in the absence of a concentration gradient; 5) Detect changes in C3b receptor expression by flow cytometry; 6) Measure superoxide generation by cytochrome c reduction assay and assess lysosomal enzyme release by enzymatic methods; 7) Include enantiomeric controls such as 8(S)-HETE and 8(R)-HETE to evaluate stereospecificity.
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| Animal Protocol |
In vivo studies of (±)8-HETE can follow the protocol below: 1) Use 6-8 week old male mice (such as CD-1 or C57BL/6 strains); 2) Induce 8-HETE production by a single topical application of TPA (20 nmol per mouse) to the back skin; 3) Euthanize animals at various time points (0, 3, 6, 12, 24, 48 hours) and collect back skin; 4) Prepare cell-free epidermal extracts and add exogenous arachidonic acid as substrate; 5) Purify metabolites by sequential thin-layer chromatography and HPLC; 6) Perform structural identification by GC-MS and ¹H-NMR spectroscopy; 7) For drug intervention studies, administer compounds by oral gavage or intraperitoneal injection (refer to formulation: 5% DMSO + 30% PEG300 + 5% Tween 80 + 60% saline), collect plasma and urine samples after 2-4 weeks of consecutive dosing for LC-MS/MS quantification.
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| ADME/Pharmacokinetics |
Metabolism / Metabolites
8-HETE is a known human metabolite of arachidonic acid. Systematic pharmacokinetic data for (±)8-HETE are limited in publicly available literature. As an endogenous arachidonic acid metabolite, its in vivo levels are regulated by multiple factors including substrate (arachidonic acid) availability and lipoxygenase/cyclooxygenase activities. Studies have shown that 8-HETE levels in plasma and urine are significantly elevated under pathological conditions such as benign prostatic hyperplasia. This compound is produced via non-enzymatic oxidation pathways and can also be generated by microsomal cytochrome P-450. Storage conditions: powder is stable for 3 years at -20°C and 2 years at 4°C; solutions are stable for 6 months at -80°C and 1 month at -20°C. The compound remains stable for several days at room temperature during shipping. |
| Toxicity/Toxicokinetics |
According to the Material Safety Data Sheet, (±)8-HETE is classified as a non-hazardous substance or mixture with no GHS hazard classification identified. This product is for research use only and is not intended for human or veterinary use. The compound is classified as a dangerous good for transport and requires additional shipping charges. Systematic toxicological data for (±)8-HETE (including acute toxicity, genotoxicity, and reproductive toxicity) are not fully reported in the available literature. As an endogenous lipid mediator, it generally does not exhibit obvious cytotoxicity at physiological concentrations. Appropriate personal protective equipment should be used during handling and storage, following standard chemical safety practices.
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| References |
[1]. Beneficial effects of eicosapentaenoic acid on the metabolic profile of obese female mice entails upregulation of HEPEs and increased abundance of enteric Akkermansia Muciniphila. Biochim Biophys Acta Mol Cell Biol Lipids. 2022 Jan; 1867(1): 159059.
[2]. Arachidonic acid metabolomic study of BPH in rats and the interventional effects of Zishen pill, a traditional Chinese medicine. J Pharm Biomed Anal . 2016 Sep 5:128:149-157. |
| Additional Infomation |
8-HETE is a HETE containing an 8-hydroxyl group and (5Z)-, (9E)-, (11Z)-, and (14Z)- double bonds. It is a mouse metabolite. It is the conjugate acid of 8-HETE(1-).
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| Molecular Formula |
C20H32O3
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|---|---|
| Molecular Weight |
320.47
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| Exact Mass |
320.235
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| CAS # |
79495-84-4
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| Related CAS # |
98462-03-4; 79495-84-4; 70968-93-3; 105500-09-2
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| PubChem CID |
11976122
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| Appearance |
Typically exists as Colorless to light yellow liquids
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| Density |
1.0±0.1 g/cm3
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| Boiling Point |
471.1±45.0 °C at 760 mmHg
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| Flash Point |
252.8±25.2 °C
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| Vapour Pressure |
0.0±2.7 mmHg at 25°C
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| Index of Refraction |
1.514
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| LogP |
5.45
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
14
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| Heavy Atom Count |
23
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| Complexity |
392
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CCCCCC=CCC=CC=CC(CC=CCCCC(=O)O)O
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| InChi Key |
NLUNAYAEIJYXRB-HEJOTXCHSA-N
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| InChi Code |
InChI=1S/C20H32O3/c1-2-3-4-5-6-7-8-9-10-13-16-19(21)17-14-11-12-15-18-20(22)23/h6-7,9-11,13-14,16,19,21H,2-5,8,12,15,17-18H2,1H3,(H,22,23)/b7-6-,10-9-,14-11-,16-13+
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| Chemical Name |
(5Z,9E,11Z,14Z)-8-hydroxyicosa-5,9,11,14-tetraenoic acid
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| Synonyms |
8-Hete; 79495-84-4; 8-hydroxy-5Z,9E,11Z,14Z-eicosatetraenoic acid; ( inverted exclamation markA)8-HETE; 70968-93-3; (5Z,9E,11Z,14Z)-8-hydroxyicosa-5,9,11,14-tetraenoic acid; (+/-)8-HETE; 5,9,11,14-Eicosatetraenoicacid, 8-hydroxy-, (5Z,9E,11Z,14Z)-;
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.1204 mL | 15.6021 mL | 31.2042 mL | |
| 5 mM | 0.6241 mL | 3.1204 mL | 6.2408 mL | |
| 10 mM | 0.3120 mL | 1.5602 mL | 3.1204 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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