| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| Other Sizes |
| Targets |
Theobromine (non-deuterated) primarily targets adenosine receptors, specifically the A1 receptor (AR1), where it acts as an antagonist. By blocking adenosine A1 receptors, theobromine stimulates the central nervous system, dilates blood vessels (vasodilation), acts as a mild diuretic, and increases heart rate. Theobromine also inhibits phosphodiesterase enzymes (PDEs), leading to increased intracellular cAMP levels and smooth muscle relaxation. The deuterated version (Theobromine-d3) shares the same mechanism but is used as an internal standard for quantification.
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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].
In vitro, theobromine (non-deuterated) inhibits adenosine A1 receptor signaling, resulting in increased neuronal activity and bronchodilation. It also inhibits phosphodiesterase, leading to smooth muscle relaxation in the bronchi and blood vessels. Theobromine has been shown to have anti-inflammatory effects in cell culture models of asthma. It also exhibits antioxidant properties by scavenging free radicals. Theobromine-d3 is not used for activity studies but serves as an internal standard for quantifying theobromine levels in cell lysates and culture media by LC-MS. |
| ln Vivo |
In vivo, theobromine (non-deuterated) has been studied for its pharmacological effects including mild central nervous system stimulation (similar to caffeine but less potent), vasodilation, and mild diuresis. It has also been investigated for potential therapeutic applications in asthma due to its bronchodilatory effects and in cardiovascular disease due to its vasodilatory properties. Theobromine is naturally present in cocoa and chocolate and is consumed orally. Theobromine-d3 is not used for in vivo activity studies; its use is limited to analytical quantification as an internal standard for LC-MS/MS.
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| Enzyme Assay |
For non-cellular assays (analytical quantification), Theobromine-d3 is prepared as a stock solution in methanol (1 mg/mL). For LC-MS/MS analysis, a calibration curve for theobromine is prepared in human plasma or urine (0.1-1000 ng/mL) with a fixed concentration of theobromine-d3 (e.g., 50 ng/mL). Sample preparation: 500 uL plasma + 50 uL internal standard + 1 mL acetonitrile for protein precipitation. After centrifugation, the supernatant is evaporated to dryness, reconstituted in 100 uL mobile phase, and injected onto a C18 column with a mobile phase of 0.1% formic acid in water and acetonitrile (gradient elution). MRM transitions: theobromine 181→138, theobromine-d3 184→141.
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| Cell Assay |
For cell-based assays, cells (e.g., neuronal cells, bronchial epithelial cells, or hepatocytes) are cultured in DMEM with 10% FBS. Cells are seeded in 6-well plates (1×10⁶ cells/well) and treated with theobromine (0.1-100 uM) for 24-72 hours. Adenosine A1 receptor signaling is assessed by measuring cAMP levels (ELISA) following forskolin stimulation. Cell viability is assessed by MTT assay. For quantification of intracellular theobromine, cells are lysed with 80% methanol containing theobromine-d3 (internal standard), and samples are analyzed by LC-MS/MS. The deuterated version (Theobromine-d3) is added as internal standard before extraction.
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| Animal Protocol |
For in vivo animal experiments, theobromine-d3 is not typically administered to animals as a drug. For pharmacokinetic studies, theobromine-d3 can be used as a tracer. Rats or mice are administered theobromine (5-20 mg/kg, oral or IV). Blood samples are collected at multiple time points (0, 0.5, 1, 2, 4, 6, 8, 12, 24 hours). Theobromine-d3 (fixed concentration) is added to plasma samples as internal standard before LC-MS/MS analysis. For tissue distribution studies, organs (brain, liver, kidney, heart) are harvested and homogenized. For diagnostic applications, theobromine-d3 is used as internal standard to quantify theobromine in biological samples.
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| ADME/Pharmacokinetics |
Theobromine-d3 has a molecular weight of 183.18 and a melting point of 345degC. The compound is soluble in DMSO (1.1 mg/mL) and slightly soluble in water (1.1 mg/mL). It has a logP of -0.8, indicating moderate hydrophilicity. The deuterium labeling provides a mass shift of +3 Da, enabling clear differentiation from non-deuterated theobromine in mass spectrometry. The compound should be stored as a powder at -20degC in a sealed, dry container, protected from moisture and light, with stability for up to 3 years.
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| Toxicity/Toxicokinetics |
Theobromine-d3 is a stable isotope-labeled compound with minimal toxicity at analytical concentrations (ng-ug per sample). The non-deuterated parent compound, theobromine, is a natural methylxanthine with low acute toxicity (LD₅0 oral rat: approximately 1,000 mg/kg). At high doses, theobromine can cause nausea, headache, and cardiac arrhythmias. However, when used as an internal standard, the amount is negligible and poses no toxicity risk. Standard laboratory safety precautions for handling organic compounds should be followed.
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| References |
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| Additional Infomation |
Theobromine-d3 is an analytical standard and research tool, not an approved drug. No clinical trials have been conducted with the deuterated version. The non-deuterated parent compound, theobromine, is a natural product consumed in cocoa and chocolate and has been investigated for potential health benefits in cardiovascular disease, asthma, and cough. It is also a minor component of some over-the-counter supplements. Theobromine-d3 is used exclusively as an internal standard for quantitative LC-MS/MS analysis of theobromine in research and quality control settings, including food analysis (determination of methylxanthines in chocolate products) and pharmacokinetic studies.
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| Molecular Formula |
C7H8N4O2
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| Molecular Weight |
180.16402053833
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| Exact Mass |
183.083
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| CAS # |
65566-69-0
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| Related CAS # |
Theobromine;83-67-0
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| PubChem CID |
16120415
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| Appearance |
White to off-white solid powder
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| LogP |
-0.8
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
13
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| Complexity |
267
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| Defined Atom Stereocenter Count |
0
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| SMILES |
CN1C(NC(=O)C2N(C([H])([H])[H])C=NC1=2)=O
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| InChi Key |
YAPQBXQYLJRXSA-FIBGUPNXSA-N
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| InChi Code |
InChI=1S/C7H8N4O2/c1-10-3-8-5-4(10)6(12)9-7(13)11(5)2/h3H,1-2H3,(H,9,12,13)/i1D3
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| Chemical Name |
3-methyl-7-(trideuteriomethyl)purine-2,6-dione
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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 | 5.5506 mL | 27.7531 mL | 55.5062 mL | |
| 5 mM | 1.1101 mL | 5.5506 mL | 11.1012 mL | |
| 10 mM | 0.5551 mL | 2.7753 mL | 5.5506 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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