| Size | Price | Stock | Qty |
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| 5mg |
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| Other Sizes |
| Targets |
N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 is a stable isotope-labeled internal standard. Its unlabeled parent compound, AAMA, is a detoxification metabolite of the genotoxic carcinogen acrylamide. AAMA itself has no known pharmacological or therapeutic targets. Instead, it serves as a major biomarker for assessing exposure to acrylamide in humans. The metabolic pathway involves the conjugation of acrylamide (which is electrophilic) with glutathione (GSH) via glutathione S-transferase (GST), followed by enzymatic conversion to a mercapturic acid derivative (N-acetyl-S-(carbamoylethyl)-L-cysteine). The formation of AAMA represents a detoxification route, removing the toxic acrylamide from the body. The labeled version is used exclusively as an internal standard to precisely quantify this biomarker, thus indirectly assessing the activity of detoxification enzymes like GSTs.
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| 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].
N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 itself is not used in in vitro activity assays. However, its unlabeled parent, AAMA, can be used as a marker. AAMA is not a bioactive molecule with an independent mechanism of action; it is simply a metabolite. In vitro studies have shown that acrylamide itself, not AAMA, is toxic. For example, in HepG2 human liver cells, acrylamide (100 uM-10 mM) induces cytotoxicity, DNA damage (Comet assay), and increased reactive oxygen species (ROS). Adding the labeled internal standard (N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3) to the culture medium does not alter these effects. Instead, it is used in the analytical LC-MS protocol to accurately measure the conversion of acrylamide to AAMA in the culture medium. This provides a quantitative measure of the detoxification capacity of the cells. This protocol is often used to test the effect of chemopreventive agents on detoxification pathways. |
| ln Vivo |
The in vivo biological activity of N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 is not characterized as it is an analytical standard. Its unlabeled parent, AAMA, is the major urinary metabolite used to monitor acrylamide exposure. In animal studies, when rats or mice are orally administered acrylamide (5-50 mg/kg), AAMA appears in the urine within hours and accounts for approximately 50-60% of the total acrylamide dose excreted. The measurement of AAMA serves as a reliable exposure biomarker. The labeled compound (d3) is used as an internal standard to quantify these levels. Furthermore, studies have shown that the levels of AAMA in urine correlate well with the degree of acrylamide-induced neurotoxicity (e.g., hind limb splay) and genotoxicity (micronucleus frequency) in these models. Thus, while the labeled compound has no activity, it is critical for quantifying the activity of the parent toxicant.
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| Enzyme Assay |
A generic non-cell-based assay for N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 (AAMA-d3) involves its use as an internal standard in an isotope dilution mass spectrometry method for urine analysis. Prepare standard stock solutions of the unlabeled AAMA in water (1 mg/mL). Prepare a separate stock solution of the internal standard (AAMA-d3) at the same concentration. Prepare calibration standards by spiking the unlabeled analyte into a blank matrix (e.g., synthetic urine) to achieve concentrations ranging from 5 ng/mL to 500 ng/mL. Add a fixed concentration of the internal standard (e.g., 100 ng/mL) to each calibration standard. Also prepare blank and double-blank samples. For sample preparation, dilute urine samples (1:1) with water. Perform solid-phase extraction (SPE) using a weak anion exchange (WAX) cartridge. Load the sample, wash with methanol, and elute with 5% ammonium hydroxide in methanol. Evaporate the eluate under nitrogen and reconstitute in mobile phase. Analyze the samples by LC-MS/MS in positive ion mode. Monitor the mass transitions: m/z 235 → 104 for AAMA, and m/z 238 → 107 for AAMA-d3. Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration.
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| Cell Assay |
The in vitro cellular assay for N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 is not for its own activity but for quantifying the metabolism of acrylamide. Culture HepG2 human hepatocarcinoma cells in DMEM with 10% FBS and antibiotics. Seed cells in 12-well plates at 5×10⁵ cells/well and allow them to attach for 24 hours. Treat the cells with different concentrations of unlabeled acrylamide (100 uM, 500 uM, 1 mM) for 24 hours. Collect the cell culture supernatant. Prepare a set of calibration standards and quality control (QC) samples by spiking known amounts of unlabeled AAMA into cell culture medium. To each sample (100 uL of supernatant), add a fixed amount of the internal standard N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 (AAMA-d3). Mix with 200 uL of acetonitrile to precipitate proteins. Centrifuge at 12,000g for 5 minutes. Transfer the supernatant to an autosampler vial. Inject the sample into the LC-MS/MS system. Quantify the concentration of AAMA in each well using the calibration curve. This protocol determines the detoxification capacity of the liver cells (how much acrylamide is converted to AAMA).
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| Animal Protocol |
A typical in vivo animal protocol for N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 involves an oral toxicokinetic study of acrylamide. Use male Sprague-Dawley rats (200-250 g, n=5 per time point). Administer a single oral dose of unlabeled acrylamide (10 mg/kg) dissolved in saline. Place the rats in individual metabolic cages to collect 24-hour urine samples. Collect blood samples via the tail vein at various time points (0, 0.5, 1, 2, 4, 8, 12, 24 h) into heparinized tubes. Centrifuge the blood to obtain plasma. For bioanalysis, mix 50 uL of plasma or urine with 50 uL of internal standard solution (containing a fixed amount of N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3). Precipitate proteins with acetonitrile. Centrifuge and inject the supernatant into the LC-MS/MS. Quantify the levels of AAMA. Calculate the pharmacokinetic parameters of AAMA formation, such as Cmax, Tmax, and AUC, which are used to describe the rate and extent of acrylamide metabolism and elimination. This method is critical for human risk assessment of acrylamide.
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| ADME/Pharmacokinetics |
N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 (AAMA-d3) itself has no pharmacologic action, so its PK is not studied. It is used as an internal standard to measure its unlabeled parent, AAMA. AAMA is a phase II metabolite of acrylamide. In humans, following dietary exposure, acrylamide is rapidly absorbed and extensively distributed. The half-life of acrylamide in blood is approximately 2-4 hours. AAMA is the major metabolite, accounting for ~40-50% of the dose, with a formation half-life of ~2 hours. AAMA has a longer elimination half-life (8-12 hours) than the parent acrylamide and is almost exclusively excreted in the urine. In kinetic studies, the area under the curve (AUC) of AAMA in urine is highly correlated with the internal dose of acrylamide. The deuterated standard is critical for accurately quantifying these parameters in mass spectrometry assays, especially given the relatively small sample volumes (e.g., 100 uL) typically used.
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| Toxicity/Toxicokinetics |
N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 (AAMA-d3) is a stable isotope-labeled research compound and is not a pharmaceutical drug. The toxicity of its unlabeled parent, AAMA, is negligible; it is a detoxification product. The toxic agent is acrylamide. Acrylamide is classified as a probable human carcinogen (Group 2A by IARC) and is genotoxic. In animal studies, acrylamide causes various tumors, neurological damage, and reproductive toxicity. The oral LD₅0 of acrylamide in rats is 150-200 mg/kg. N-Acetyl-S-(carbamoylethyl)-L-cysteine (AAMA) has no known carcinogenic, mutagenic, or toxic properties. For laboratory handling of the labeled AAMA-d3, standard chemical safety practices should be used. However, since it is a metabolite of a toxic compound, it should be handled with care, and any potential contamination with acrylamide should be ruled out. The compound should be stored at -20degC.
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| References | |
| Additional Infomation |
N-Acetyl-S-(carbamoylethyl)-L-cysteine-d3 (AAMA-d3) is the stable isotope-labeled version of N-Acetyl-S-(carbamoylethyl)-L-cysteine (AAMA), the major urinary metabolite of the genotoxic carcinogen acrylamide. It is intended for research use only as a sensitive and specific internal standard for the quantification of human exposure to acrylamide by LC-MS/MS. Acrylamide is formed from the reaction of asparagine and reducing sugars during high-temperature cooking (e.g., frying, baking) in a wide range of foods, including potato chips, coffee, and cereals. Measuring AAMA levels in urine is the most common method for biomonitoring and assessing human exposure in nutritional and occupational settings. The labeled compound (d3) corrects for variations in sample preparation and analysis, providing accurate risk assessment data for this widespread food contaminant.
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| Molecular Formula |
C8H11D3N2O4S
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|---|---|
| Molecular Weight |
237.29
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| Exact Mass |
237.086
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| CAS # |
1795786-57-0
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| PubChem CID |
71312851
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| Appearance |
Typically exists as solid at room temperature
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| Melting Point |
99-102°C
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| LogP |
-1.3
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| Hydrogen Bond Donor Count |
3
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| Hydrogen Bond Acceptor Count |
5
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
15
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| Complexity |
257
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| Defined Atom Stereocenter Count |
1
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| SMILES |
[C@H](C(=O)O)(NC(=O)C([H])([H])[H])CSCCC(=O)N
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| InChi Key |
GGBCHNJZQQEQRX-FYFSCIFKSA-N
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| InChi Code |
InChI=1S/C8H14N2O4S/c1-5(11)10-6(8(13)14)4-15-3-2-7(9)12/h6H,2-4H2,1H3,(H2,9,12)(H,10,11)(H,13,14)/t6-/m0/s1/i1D3
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| Chemical Name |
(2R)-3-(3-amino-3-oxopropyl)sulfanyl-2-[(2,2,2-trideuterioacetyl)amino]propanoic 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 | 4.2143 mL | 21.0713 mL | 42.1425 mL | |
| 5 mM | 0.8429 mL | 4.2143 mL | 8.4285 mL | |
| 10 mM | 0.4214 mL | 2.1071 mL | 4.2143 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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