| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| Other Sizes |
| Targets |
Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 is a stable isotope-labeled internal standard. Its unlabeled parent, acetaldehyde, is a highly reactive, small aldehyde with significant toxicological and pharmacological targets. Acetaldehyde is the primary metabolite of ethanol (via alcohol dehydrogenase, ADH) and is responsible for many of the toxic and carcinogenic effects of alcohol consumption. Its primary biological targets include: (1) DNA: acetaldehyde forms DNA adducts, primarily with guanine residues (e.g., N2-ethylidene-dG), leading to mutations and carcinogenesis. (2) Proteins: it forms protein adducts, interfering with enzyme function (e.g., inhibiting aldehyde dehydrogenase, ALDH2, leading to acetaldehyde accumulation and the “flushing” response). (3) Collagen: it stimulates collagen production, contributing to alcoholic liver fibrosis (cirrhosis). The labeled DNPH derivative is used exclusively for analytical quantification, not for studying these toxic mechanisms directly.
|
|---|---|
| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as quantitative tracers while the drugs were being developed. Because deuteration may have an effect on a drug's pharmacokinetics and metabolic properties, it is a cause for concern [1].
The in vitro biological activity of Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 is not characterized; it is used as an analytical internal standard. Its unlabeled parent, acetaldehyde, exhibits significant biological activity in vitro. In cultured human bronchial epithelial cells (BEAS-2B), acetaldehyde (50-500 uM) induces DNA damage (as measured by the Comet assay), increases the frequency of micronuclei, and upregulates the expression of pro-inflammatory cytokines (IL-8, TNF-alpha). In HepG2 human hepatoma cells, acetaldehyde (100-1,000 uM) depletes glutathione (GSH) levels, increases reactive oxygen species (ROS) production, and induces apoptosis. In primary rat hepatocytes, acetaldehyde inhibits the activity of ALDH2 (the mitochondrial isoform) with an IC₅0 of approximately 30-50 uM. Acetaldehyde also inhibits the repair of DNA alkylation damage. In addition, it is mutagenic in the Ames test (Salmonella typhimurium TA100) in the presence of metabolic activation (S9 mix), inducing frameshift and base-pair substitution mutations. The labeled derivative is used as an internal standard to quantify acetaldehyde in these in vitro cultures after derivatization. |
| ln Vivo |
The in vivo activity of Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 is not evaluated; it is used as a reference standard. Its unlabeled parent, acetaldehyde, is a highly toxic compound. In animals, acute acetaldehyde administration (50-200 mg/kg i.p.) produces flushing, tachycardia, hypotension, nausea, and respiratory depression, closely mimicking the symptoms of alcohol intolerance (disulfiram-like reaction). Chronic acetaldehyde exposure in animal models (e.g., via inhalation or in drinking water) leads to hepatocellular carcinoma (liver cancer), squamous cell carcinoma of the upper aerodigestive tract, and fetal developmental toxicity (in utero exposure). Acetaldehyde is classified by IARC as Group 1 (“carcinogenic to humans”) based on sufficient evidence of human carcinogenicity (cancer of the esophagus, liver, and breast). In humans, the acetaldehyde content in tobacco smoke and the endogenous production of acetaldehyde from ethanol metabolism (especially in individuals with the ALDH2*2 polymorphism, who accumulate acetaldehyde) are major risk factors for cancer. The labeled acetaldehyde-DNPH derivative is critical for accurate quantification of acetaldehyde in biological matrices (blood, urine, exhaled breath) for exposure biomonitoring and alcohol metabolism studies.
|
| Enzyme Assay |
A generic non-cell-based assay for Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 involves its use as an internal standard in an HPLC-UV or LC-MS/MS method for quantifying acetaldehyde in air samples after DNPH derivatization. For air sampling, pass a known volume of air (e.g., 100 L) through a DNPH-coated silica gel cartridge at a flow rate of 1 L/min for 60 minutes. The DNPH reacts with acetaldehyde to form the corresponding hydrazone derivative. Extract the cartridge with 5 mL of acetonitrile. For calibration, prepare standard solutions of unlabeled acetaldehyde-DNPH derivative (acetaldehyde 2,4-dinitrophenylhydrazone) in acetonitrile (0.1-10 ug/mL). Add a fixed concentration of Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 (e.g., 2 ug/mL) as the internal standard to each calibration standard. Analyze by HPLC with UV detection at 360 nm (DNPH derivatives have strong absorbance at 360 nm) or by LC-MS/MS in negative ion mode. Monitor the mass transitions: m/z 223 → 152 for unlabeled acetaldehyde-DNPH, and m/z 226 → 155 for the deuterated internal standard. Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration. This method is used for environmental monitoring of acetaldehyde in ambient air (workplace exposure, indoor air quality).
|
| Cell Assay |
A standard in vitro cell-based protocol for acetaldehyde analysis involves measuring acetaldehyde levels in cell culture medium after exposure to ethanol (to study alcohol metabolism). Culture human hepatoma HepG2 cells in DMEM supplemented with 10% FBS and 1% penicillin/streptomycin at 37degC in a 5% CO2 incubator. Seed cells in 6-well plates at 2×10⁵ cells/well. Treat cells with increasing concentrations of unlabeled ethanol (0, 10, 50, 100, 250, 500 mM) for 6, 12, and 24 hours. Include control wells with the ALDH2 inhibitor disulfiram (1 uM) to block acetaldehyde metabolism if desired. At each time point, collect the culture medium (1 mL). Immediately add 100 uL of a DNPH derivatization reagent solution (30 mM DNPH in 2 M HCl) to the medium sample to trap acetaldehyde as the hydrazone derivative. Incubate at room temperature for 60 minutes. Extract the derivatized sample with 2 mL of ethyl acetate. Evaporate the organic layer under nitrogen and reconstitute in 200 uL of acetonitrile. Add a fixed amount of Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 as internal standard. Analyze by LC-MS/MS. Quantify the concentration of acetaldehyde produced in the culture medium. This protocol is used to study the effects of genetic polymorphisms and drug interactions on acetaldehyde formation from ethanol.
|
| Animal Protocol |
A typical in vivo animal protocol for Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 is used in a toxicokinetic study of ethanol and acetaldehyde in rats. Use male Sprague-Dawley rats (250-300 g, n = 5-6 per group). Administer a single oral dose of unlabeled ethanol (2 g/kg, 20% v/v in water) via gavage. Include a group treated with disulfiram (50 mg/kg, i.p.) 30 minutes before ethanol administration to inhibit ALDH2. Collect blood samples via tail vein at various time points (0, 0.5, 1, 2, 4, 6, 8, 12, 24 h). Immediately place the blood samples (50 uL) into chilled tubes containing 10 uL of a DNPH derivatization solution (30 mM DNPH in 2 M HCl) to trap any acetaldehyde present. Keep the tubes on ice for 60 minutes, then extract with ethyl acetate. Add Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 as an internal standard to the extract. Analyze by LC-MS/MS. Quantify the blood acetaldehyde concentration (ng/mL). Also measure ethanol levels in the plasma by a standard enzymatic (ADH) assay. Compare the acetaldehyde levels in the disulfiram-treated group (where acetaldehyde accumulates) vs. the control group. Calculate the ratio of acetaldehyde to ethanol (AUCAcH/AUCEtOH) as a measure of the rate of ethanol metabolism to acetaldehyde and the clearance of acetaldehyde.
|
| ADME/Pharmacokinetics |
Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 is an analytical standard, not a drug. Its unlabeled parent, acetaldehyde, is a highly toxic and carcinogenic compound. Acetaldehyde is classified by the International Agency for Research on Cancer (IARC) as Group 1 (“carcinogenic to humans”), based on sufficient evidence for esophageal cancer and breast cancer in humans, as well as strong mechanistic evidence for DNA adduct formation and mutagenicity. The acute toxicity (oral LD₅0 in rats) is approximately 1,930 mg/kg for acetaldehyde (moderate acute toxicity). Acute exposure to high concentrations can cause severe eye, skin, and respiratory tract irritation, as well as central nervous system depression. Long-term exposure (chronic) is associated with liver cirrhosis, liver cancer, and damage to the upper respiratory tract and gastrointestinal tract. For laboratory handling of the deuterated DNPH derivative, the compound is a solid or crystalline powder and is much less volatile and less toxic than acetaldehyde itself. However, standard laboratory safety precautions should be followed: wear PPE (gloves, lab coat, goggles), work in a fume hood to avoid dust, and wash hands thoroughly after handling. Store at -20degC or 2-8degC in a tightly sealed container, protected from light. For research use only, not for human consumption.
|
| References | |
| Additional Infomation |
Acetaldehyde 2,4-Dinitrophenylhydrazone-3,5,6-d3 is the stable isotope-labeled version of acetaldehyde 2,4-dinitrophenylhydrazone (acetaldehyde-DNPH), the derivatized product of the volatile aldehyde acetaldehyde. Acetaldehyde is a widespread environmental pollutant, a major component of tobacco smoke, and the primary metabolite of ethanol (alcohol) in humans, formed by the action of alcohol dehydrogenase (ADH). It is a highly reactive compound and a known human carcinogen (IARC Group 1), implicated in the development of esophageal cancer, hepatocellular carcinoma (liver cancer), and breast cancer. The labeled DNPH derivative is intended for research use as an internal standard for the accurate quantification of acetaldehyde in air, water, biological fluids (blood, urine), and food samples by HPLC-UV or LC-MS/MS following derivatization with 2,4-dinitrophenylhydrazine (DNPH). This compound is an essential analytical tool for environmental monitoring (indoor/outdoor air quality, workplace exposure), occupational health studies, and research on alcohol metabolism and carcinogenesis. For research use only, not for diagnostic or therapeutic applications.
|
| Molecular Formula |
C8H5D3N4O4
|
|---|---|
| Molecular Weight |
227.19
|
| Exact Mass |
227.073
|
| CAS # |
259824-51-6
|
| PubChem CID |
70553
|
| Appearance |
Light yellow to yellow solid powder
|
| Density |
1.5±0.1 g/cm3
|
| Boiling Point |
382.5±42.0 °C at 760 mmHg
|
| Melting Point |
147-149°C
|
| Flash Point |
185.2±27.9 °C
|
| Vapour Pressure |
0.0±0.9 mmHg at 25°C
|
| Index of Refraction |
1.629
|
| LogP |
2.83
|
| Hydrogen Bond Donor Count |
1
|
| Hydrogen Bond Acceptor Count |
6
|
| Rotatable Bond Count |
2
|
| Heavy Atom Count |
16
|
| Complexity |
295
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
[2H]C1=C(C(=C(C(=C1N/N=C/C)[N+](=O)[O-])[2H])[N+](=O)[O-])[2H]
|
| InChi Key |
ONBOQRNOMHHDFB-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C8H8N4O4/c1-2-9-10-7-4-3-6(11(13)14)5-8(7)12(15)16/h2-5,10H,1H3
|
| Chemical Name |
N-(ethylideneamino)-2,4-dinitroaniline
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 4.4016 mL | 22.0080 mL | 44.0160 mL | |
| 5 mM | 0.8803 mL | 4.4016 mL | 8.8032 mL | |
| 10 mM | 0.4402 mL | 2.2008 mL | 4.4016 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
