| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg | |||
| Other Sizes |
| Targets |
Ibuprofen-d4 targets the same enzymes as the unlabeled parent drug, specifically cyclooxygenase-1 (COX-1) and cyclooxygenase-2 (COX-2). Ibuprofen acts as a selective COX-1 inhibitor with an IC50 value of 13 microM. By inhibiting COX enzymes, ibuprofen blocks the synthesis of prostaglandins, thereby exerting anti-inflammatory, analgesic, and antipyretic effects. Ibuprofen-d4 is not used for activity assays but for quantification.
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| ln Vitro |
Drug compounds have included stable heavy isotopes of carbon, hydrogen, and other elements, mostly as tracers for quantification throughout the drug development process. Due to its potential to alter the pharmacokinetic and metabolic characteristics of medications, deuteration has drawn attention[1].
Ibuprofen-d4 is not intended for use in activity assays, but the unlabeled ibuprofen inhibits COX-1 and COX-2 in cell-free enzyme assays. Purified ovine COX-1 or human recombinant COX-2 is incubated with arachidonic acid (substrate) in the presence of varying concentrations of ibuprofen. The production of prostaglandin E2 (PGE2) is measured by ELISA. Ibuprofen-d4 is used as an internal standard for quantification in such assays. |
| ln Vivo |
In cellular models, ibuprofen (unlabeled) inhibits LPS-induced PGE2 production in macrophages and other inflammatory cells. The compound suppresses cell proliferation, inhibits angiogenesis, and induces apoptosis in various cancer cell lines. Ibuprofen-d4 is not used in these bioactivity assays but serves as an internal standard for measuring ibuprofen concentrations in cell lysates and culture media.
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| Enzyme Assay |
COX-1 enzyme inhibition assays are performed using purified ovine COX-1 (or human recombinant). The enzyme is incubated with arachidonic acid (10 microM) and varying concentrations of ibuprofen (0.1-1000 microM) in Tris-HCl buffer (pH 7.4). After 10 minutes, the reaction is stopped with HCl, and the PGE2 produced is quantified by ELISA. The IC50 value is calculated from the dose-response curve. Ibuprofen-d4 is used as an internal standard for LC-MS/MS validation of the assay.
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| Cell Assay |
Inflammatory cells (e.g., RAW264.7 macrophages) are seeded in 96-well plates and treated with ibuprofen (1-1000 microM) for 1 hour, followed by LPS (1 microg/mL) stimulation for 24 hours. The culture supernatant is collected, and PGE2 concentration is measured by ELISA. Cell viability is assessed by MTT to exclude cytotoxicity. Ibuprofen-d4 is used as an internal standard for drug quantification in cell-based uptake studies via LC-MS/MS.
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| Animal Protocol |
In a rat model of carrageenan-induced paw edema, ibuprofen-d4 is administered orally or intraperitoneally (10-50 mg/kg) as a tracer along with unlabeled ibuprofen. Blood samples are collected at various time points (0-24 hours), and plasma is analyzed by LC-MS/MS using Ibuprofen-d4 as the internal standard for quantification. Edema volume is measured using plethysmography. Other models include adjuvant-induced arthritis and fever models in rodents.
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| ADME/Pharmacokinetics |
Ibuprofen is well absorbed after oral administration, reaching peak plasma concentrations within 1-2 hours. It is highly protein-bound (>99%) and has a half-life of approximately 2 hours. The deuterium label (d4) in Ibuprofen-d4 does not alter the pharmacokinetic properties of the drug, making it an ideal internal standard for bioequivalence and bioavailability studies. Ibuprofen-d4 is commonly used in LC-MS/MS methods to quantify ibuprofen levels.
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| Toxicity/Toxicokinetics |
Ibuprofen-d4 is used in trace amounts as an analytical standard and does not contribute to toxicity. The unlabeled ibuprofen has a well-established safety profile. Common adverse effects include gastrointestinal irritation, nausea, and headache. At high doses or with long-term use, there is a risk of gastric ulcers, renal impairment, and cardiovascular events. The deuterium label does not alter the toxicity profile of the compound.
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| References |
[1]. Russak EM, et al. Impact of Deuterium Substitution on the Pharmacokinetics of Pharmaceuticals. Ann Pharmacother. 2019 Feb;53(2):211-216.
[2]. Noreen Y, et al. Development of a radiochemical cyclooxygenase-1 and -2 in vitro assay for identification of natural products as inhibitors of prostaglandin biosynthesis. J Nat Prod. 1998 Jan;61(1):2-7. [3]. Hassan Akrami, et al. Inhibitory effect of ibuprofen on tumor survival and angiogenesis in gastric cancer cell. Tumour Biol. 2015 May36(5):3237-43. [4]. Nathan D Pennock, et al. Ibuprofen supports macrophage differentiation, T cell recruitment, and tumor suppression in a model of postpartum breast cancer. J Immunother Cancer. 2018 Oct 16(1):98. [5]. M W Konstan, et al. Ibuprofen attenuates the inflammatory response to Pseudomonas aeruginosa in a rat model of chronic pulmonary infection. Implications for antiinflammatory therapy in cystic fibrosis. Am Rev Respir Dis. 1990 Jan141(1):186-92. |
| Additional Infomation |
Ibuprofen is a widely available over-the-counter and prescription NSAID approved for the treatment of pain, fever, and inflammation. Ibuprofen-d4 is a research-grade stable isotope-labeled compound used exclusively as an internal standard for analytical method development (e.g., LC-MS/MS, GC-MS) in pharmacokinetic, bioequivalence, and forensic toxicology studies. It is not intended for therapeutic use.
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| Molecular Formula |
C13H14D4O2
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|---|---|
| Molecular Weight |
210.31
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| Related CAS # |
Ibuprofen;15687-27-1
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| Appearance |
Typically exists as solid at room temperature
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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 | 4.7549 mL | 23.7744 mL | 47.5489 mL | |
| 5 mM | 0.9510 mL | 4.7549 mL | 9.5098 mL | |
| 10 mM | 0.4755 mL | 2.3774 mL | 4.7549 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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