| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Ethylglucuronide-d5 is a stable isotope-labeled internal standard that mimics the activity of its non-labeled parent, ethyl glucuronide (EtG). The primary target of EtG is its use as a biomarker for ethanol (alcohol) consumption. It is not a pharmacologically active substance with a classical drug target, but rather serves as an indicator of recent drinking. In the body, it is generated in the liver as a direct conjugate of ethanol and glucuronic acid via UDP-glucuronosyltransferase (UGT) enzymes. While EtG itself is a metabolite, it has been recently identified as a ligand for the immune receptor Toll-like Receptor 4 (TLR4). In vitro, it has been shown to act as an agonist of TLR4, potentially contributing to alcohol-induced neuroinflammation. The labeled version (d5) is used as a tracer to study EtG without altering the analytical signal.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers that influence measurement during the drug development process. It's possible that the pharmacokinetics and functional range of medications contribute to the concern over mutagenesis [1].
The in vitro biological activity of Ethylglucuronide-d5 is presumed to be the same as its unlabeled parent, ethyl glucuronide (EtG). In cell-based assays, EtG has been shown to act as a weak agonist of Toll-like receptor 4 (TLR4). In microglial cell lines (e.g., BV-2 cells), treatment with EtG (1-100 uM) led to the dose-dependent activation of the NF-kappaB pathway and increased the expression of pro-inflammatory cytokines such as TNF-alpha, IL-1beta, and IL-6. This suggests that EtG may contribute to the neuroinflammatory effects associated with chronic alcohol consumption. However, the potency of EtG is significantly lower than that of its parent compound, ethanol. EtG is primarily used as a diagnostic biomarker for alcohol intake, rather than as a biologically active molecule. Ethylglucuronide-d5 is used as an internal standard to accurately measure the concentrations of EtG in such cell culture experiments. |
| ln Vivo |
Ethylglucuronide-d5 itself is not used for in vivo activity studies. The in vivo activity of its non-labeled counterpart, ethyl glucuronide (EtG), is primarily as a biomarker of alcohol intake. When ethanol is consumed, a small fraction (0.02-0.1%) is metabolized to EtG by UDP-glucuronosyltransferase enzymes in the liver. EtG is then distributed into the bloodstream and eliminated into the urine. Unlike ethanol, which is rapidly eliminated, EtG has a longer detection window. In humans, it can be detected in urine for up to 80 hours after the complete elimination of ethanol from the blood. EtG also accumulates in hair, where it serves as a marker for long-term alcohol exposure (weeks to months). In animal studies (rats), intravenous administration of EtG (10 mg/kg) has been shown to have no obvious pharmacological effects such as sedation or motor incoordination. Ethylglucuronide-d5 is used as an internal standard in these studies to quantify EtG concentrations.
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| Enzyme Assay |
A generic non-cell-based assay for Ethylglucuronide-d5 involves its use as an internal standard in an isotope dilution mass spectrometry method. First, prepare a standard stock solution of unlabeled Ethyl glucuronide (EtG) in methanol or water at 1 mg/mL. Prepare a separate stock solution of the internal standard (Ethylglucuronide-d5) at the same concentration. Prepare calibration standards by spiking the unlabeled analyte into a blank matrix (e.g., synthetic urine or blank human urine) to achieve concentrations ranging from 0.05 ug/mL to 10 ug/mL. Add a fixed concentration of the internal standard (e.g., 0.5 ug/mL) to each calibration standard. Also prepare blank and double-blank samples. For sample preparation, dilute the samples with water and load onto a solid-phase extraction (SPE) cartridge (e.g., mixed-mode anion exchange). After washing, elute the analytes with a suitable solvent. Evaporate and reconstitute in mobile phase. Analyze the samples by LC-MS/MS in negative ion mode. Monitor the mass transitions: m/z 221 → 85 for EtG, and m/z 226 → 90 for EtG-d5. Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration.
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| Cell Assay |
Ethylglucuronide-d5 is an analytical internal standard, and a standard cell-based assay for its unlabeled parent involves measuring TLR4 activation. Culture BV-2 mouse microglial cells in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin. Seed the cells in 6-well plates at a density of 5×10⁵ cells per well and allow them to grow to 80% confluency. Treat the cells with various concentrations (0.1, 1, 10, 50, 100 uM) of unlabeled ethyl glucuronide (EtG) or LPS (positive control, 1 ug/mL) for 6-24 hours. After the treatment, collect the cell culture supernatant for cytokine measurement (e.g., TNF-alpha, IL-6 ELISA). Lyse the cells to extract total RNA for qRT-PCR analysis of inflammatory gene expression (e.g., Il-1beta, Tnf-alpha, Il-6). For validation, collect a separate set of culture media samples, add Ethylglucuronide-d5 as the internal standard, and analyze by LC-MS/MS to determine the precise concentration of EtG present in the cell culture medium during the experiment, ensuring accuracy of the dose-response relationship.
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| Animal Protocol |
A typical in vivo animal protocol for Ethylglucuronide-d5 involves a study of alcohol consumption in rats. Use male Wistar rats (200-250 g). Administer a single dose of ethanol (2-4 g/kg) via oral gavage to mimic human binge drinking. Include a control group that receives an equal volume of water. After dosing, place the rats in individual metabolic cages for 24-hour urine collection. Collect blood samples from the tail vein at various time points (0, 1, 2, 4, 8, 12, 24 h). Determine blood alcohol concentration (BAC) using an enzymatic assay or a breathalyzer. For EtG analysis, centrifuge the urine and plasma samples. Add a fixed amount of Ethylglucuronide-d5 as the internal standard to each sample. Process the samples for LC-MS/MS analysis. Calculate the total EtG excreted in urine over 24 hours. Correlate the EtG levels with the BAC and the time since the last dose. This protocol is essential for validating the use of EtG as a long-term alcohol consumption biomarker.
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| ADME/Pharmacokinetics |
Ethylglucuronide-d5 is a stable isotope-labeled internal standard. Its non-labeled parent, ethyl glucuronide (EtG), is a phase-II metabolite of ethanol. Pharmacokinetically, EtG is produced in the liver primarily by UDP-glucuronosyltransferase (UGT) enzymes, particularly UGT2B7. After ethanol consumption, plasma levels of EtG peak later (2-4 hours) than ethanol (0.5-1 hour) and remain elevated for a much longer period. The half-life of EtG in blood and urine is approximately 2-3 hours. It is distributed primarily in the blood and excreted exclusively via renal filtration, with no active secretion. There is no enterohepatic circulation. The labeling with deuterium (d5) in Ethylglucuronide-d5 does not alter its physicochemical or PK properties, making it an ideal internal standard for the accurate determination of these parameters via mass spectrometry, even at low concentrations, which is crucial for its clinical application as an abstinence marker.
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| Toxicity/Toxicokinetics |
Ethylglucuronide-d5 is not intended for human use and is classified as a research chemical. The toxicity of its unlabeled parent, ethyl glucuronide (EtG), is considered minimal. EtG is an endogenous metabolite; it is the direct conjugate of ethanol and glucuronic acid. Because EtG is produced naturally as part of ethanol metabolism, it does not exhibit significant toxicity. Its primary use is as a biomarker for alcohol consumption. Even at high concentrations, it is not known to cause adverse physiological effects. However, EtG is a known TLR4 agonist, and research suggests it might contribute to low-grade neuroinflammation following chronic alcohol consumption. For laboratory personnel, the compound is considered low-hazard. Standard procedures for handling chemicals, such as wearing gloves and a lab coat, are sufficient. It should be stored at 4degC in a sealed container, away from moisture.
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| References | |
| Additional Infomation |
Ethylglucuronide-d5 (EtG-d5) is the stable isotope-labeled (deuterated) version of ethyl glucuronide, a minor direct metabolite of ethanol. It is used exclusively as an internal standard for the precise quantification of ethyl glucuronide (EtG) in biological specimens (urine, blood, hair) by LC-MS/MS. EtG is a specific and sensitive biomarker for alcohol intake because it is produced only after ethanol consumption. It can detect alcohol use for up to 80 hours after complete elimination of ethanol, making it valuable for abstinence monitoring in clinical settings (e.g., for liver transplant candidates) and in forensic toxicology (e.g., driver‘s license reinstatement). It also accumulates in hair, allowing detection of chronic alcohol consumption over months. Ethylglucuronide-d5 is a research compound and not for diagnostic or therapeutic use.
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| Molecular Formula |
C8H9D5O7
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|---|---|
| Molecular Weight |
227.22
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| Exact Mass |
227.105
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| CAS # |
1135070-98-2
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| Related CAS # |
Ethyl glucuronide;17685-04-0
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| PubChem CID |
71316593
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| Appearance |
White to off-white solid powder
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| Melting Point |
145-147°C (dec.)
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| Flash Point |
9℃
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| LogP |
-1.4
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
7
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| Rotatable Bond Count |
3
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| Heavy Atom Count |
15
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| Complexity |
231
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| Defined Atom Stereocenter Count |
5
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| SMILES |
[2H]C([2H])([2H])C([2H])([2H])O[C@H]1[C@@H]([C@H]([C@@H]([C@H](O1)C(=O)O)O)O)O
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| InChi Key |
IWJBVMJWSPZNJH-XTJTXKPGSA-N
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| InChi Code |
InChI=1S/C8H14O7/c1-2-14-8-5(11)3(9)4(10)6(15-8)7(12)13/h3-6,8-11H,2H2,1H3,(H,12,13)/t3-,4-,5+,6-,8+/m0/s1/i1D3,2D2
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| Chemical Name |
(2S,3S,4S,5R,6R)-3,4,5-trihydroxy-6-(1,1,2,2,2-pentadeuterioethoxy)oxane-2-carboxylic acid
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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 | 4.4010 mL | 22.0051 mL | 44.0102 mL | |
| 5 mM | 0.8802 mL | 4.4010 mL | 8.8020 mL | |
| 10 mM | 0.4401 mL | 2.2005 mL | 4.4010 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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