| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
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| Other Sizes |
| Targets |
Not applicable; Theobromine-d6 is primarily a stable isotope-labeled internal standard for analytical quantification. The unlabeled parent compound theobromine is an adenosine A1 receptor antagonist with IC₅0s of 200-280 microM in radioligand binding assays using rat brain membranes. Theobromine also increases AMPK phosphorylation, inhibits adipocyte differentiation and ERK/JNK phosphorylation, and reduces IL-6 and TNF-alpha production in 3T3-L1 preadipocytes. However, the deuterated version serves as an internal standard for quantifying theobromine, not as a pharmacological agent targeting these pathways.
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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].
Theobromine-d6 is intended for use as an internal standard and does not directly exert pharmacological effects in vitro. The unlabeled theobromine (1-1000 uM) increases AMPK phosphorylation and inhibits adipocyte differentiation, ERK and JNK phosphorylation, and IL-6 and TNF-alpha production in 3T3-L1 preadipocytes cultured in differentiation medium. Theobromine (150 microg/ml) increases AMPK phosphorylation and inhibits these inflammatory and differentiation markers. It also inhibits decreases in renal cortex SIRT1 activity and increases in NADPH oxidase-dependent ROS production in renal cells. |
| ln Vivo |
In vivo, Theobromine-d6 is used as an internal standard and is not typically administered as a test compound. The unlabeled theobromine (5 mg/kg per day) inhibits decreases in renal cortex SIRT1 activity and increases in NADPH oxidase-dependent reactive oxygen species (ROS) production, as well as reduces kidney hypertrophy and albuminuria in a spontaneously hypertensive rat model of streptozotocin-induced diabetes. Theobromine also demonstrates toxic effects in dogs with an LD₅0 value of 250 to 500 mg/kg. It exhibits anti-inflammatory and cardiovascular benefits in various in vivo models.
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| Enzyme Assay |
For quantitative LC-MS/MS analysis: Prepare a stock solution of Theobromine-d6 in DMF or DMSO at 10 mg/mL. Spike a known amount (e.g., 10-100 ng/mL) into biological samples (plasma, urine, tissue homogenates) as an internal standard. Protein precipitation with acetonitrile or methanol is followed by centrifugation. Separate on a C18 reverse-phase column using a mobile phase of 0.1% formic acid in water and acetonitrile. Detect using MRM transitions: for theobromine m/z 181 → 138; for Theobromine-d6 m/z 187 → 144 (mass shift of +6 due to six deuterium atoms). Quantify by isotope dilution method against a calibration curve.
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| Cell Assay |
For cell-based quantification of theobromine, cells (e.g., 3T3-L1 preadipocytes) are cultured in differentiation medium. Harvest cells at various time points, wash with PBS, and lyse cells in methanol/water. Add Theobromine-d6 as an internal standard at a known concentration (e.g., 50 ng/mL). Centrifuge to remove debris, filter, and analyze the supernatant by LC-MS/MS as described in the assay protocol above. Quantify endogenous theobromine levels by comparing the peak area ratio of theobromine to Theobromine-d6 against a calibration curve. This method corrects for matrix effects, extraction efficiency, and ion suppression.
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| Animal Protocol |
For in vivo pharmacokinetic studies, administer theobromine to animals (e.g., rats, mice) by oral gavage or intravenous injection at specific doses (e.g., 5 mg/kg). Collect blood samples at multiple time points (0, 0.5, 1, 2, 4, 6, 8, 12, 24 h) into heparinized tubes. Centrifuge to obtain plasma. Add Theobromine-d6 internal standard (e.g., 50-100 ng/mL) to each plasma sample. Precipitate proteins with acetonitrile, centrifuge, and analyze supernatant by LC-MS/MS. Calculate pharmacokinetic parameters including Cmax, Tmax, AUC, half-life, and bioavailability using non-compartmental analysis. This method accurately quantifies theobromine without endogenous interference.
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| ADME/Pharmacokinetics |
Theobromine-d6: Molecular formula C₇H2D₆N4O2. Molecular weight: 186.20 g/mol (unlabeled theobromine 180.16). Purity: ≥99% deuterated forms (d1-d6). Appearance: A crystalline solid. Melting point: ~357degC. UV λmax: 273 nm. Solubility: DMF 10 mg/ml, DMSO 10 mg/ml, Ethanol 1 mg/ml, PBS (pH 7.2) 1 mg/ml. Storage: Store powder at -20degC. Shipping: Wet ice in continental US; may vary elsewhere. Protect from light and moisture. It is a controlled substance and not for sale in certain territories.
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| Toxicity/Toxicokinetics |
Theobromine-d6 has low toxicity for analytical use at typical working concentrations. The unlabeled parent compound theobromine is toxic to dogs with an LD₅0 value of 250 to 500 mg/kg, but it is generally regarded as safe for human consumption in chocolate and cocoa products at normal dietary levels. Theobromine can cause adverse effects in humans at very high doses (e.g., theobromine poisoning), but the deuterated internal standard is used at trace levels (ng/mL to ug/mL) for analytical purposes, where it is considered safe. Standard laboratory safety precautions should be followed when handling the pure compound: wear gloves, lab coat, and safety glasses. Avoid inhalation of powder and contact with skin/eyes. Consult SDS for detailed safety information. Not for human therapeutic use.
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| References |
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| Additional Infomation |
Theobromine-D6 is an oxopurine.
Theobromine is a methylxanthine alkaloid and derivative of caffeine that has been found in cocoa beans and has diverse biological activities. Theobromine-d6 (3,7-Dimethylxanthine-d6) is a synthetic deuterated form where the methyl groups are labeled with deuterium. The compound is supplied as a high-purity analytical standard for research use only. It is not approved for human therapeutic use. In addition to its role as an internal standard, theobromine exhibits effects on adenosine receptors, anti-inflammatory properties, and potential cardiovascular benefits. The deuterated version is also available as a crystalline solid with a minimum purity of 99% deuterated forms (d1-d6). The product is a controlled substance and not for sale in certain territories. For research use only; we do not sell to patients. |
| Molecular Formula |
C7H2D6N4O2
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|---|---|
| Molecular Weight |
186.20
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| Exact Mass |
186.102
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| CAS # |
117490-40-1
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| Related CAS # |
Theobromine;83-67-0
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| PubChem CID |
53442231
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| Appearance |
White to off-white solid powder
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| Melting Point |
>300ºC
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| Index of Refraction |
1.737
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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 |
[2H]C([2H])([2H])N1C=NC2=C1C(=O)NC(=O)N2C([2H])([2H])[2H]
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| InChi Key |
YAPQBXQYLJRXSA-WFGJKAKNSA-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)/i1D3,2D3
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| Chemical Name |
3,7-bis(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) |
DMSO: 2.86 mg/mL (15.36 mM)
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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 | 5.3706 mL | 26.8528 mL | 53.7057 mL | |
| 5 mM | 1.0741 mL | 5.3706 mL | 10.7411 mL | |
| 10 mM | 0.5371 mL | 2.6853 mL | 5.3706 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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