| Size | Price | Stock | Qty |
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| 1mg |
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| Other Sizes |
| Targets |
Benzydamine N-Oxide does not directly target biological receptors; it is an inactive metabolite of the parent drug. The parent compound, benzydamine, is a locally-acting NSAID that exerts its anti-inflammatory, analgesic, and antipyretic effects by inhibiting the production of pro-inflammatory cytokines (TNF-alpha, IL-1beta, IL-6) and the activation of NF-kappaB. It also stabilizes cell membranes, inhibits the release of histamine and other inflammatory mediators, and has local anesthetic properties. Benzydamine N-Oxide has significantly reduced pharmacological activity and is used primarily as a marker compound to study the oxidative metabolism of benzydamine.
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| ln Vitro |
In vitro, Benzydamine N-Oxide is formed when benzydamine is incubated with liver microsomes or recombinant CYP enzymes (particularly CYP3A4) in the presence of NADPH. The compound itself has minimal anti-inflammatory or analgesic activity. It is used as an internal standard in LC-MS/MS methods for the quantification of benzydamine in biological matrices (plasma, urine, tissue homogenates). Benzydamine N-Oxide has a distinct mass-to-charge ratio (m/z) from benzydamine and is stable under analytical conditions, making it ideal for assay calibration. The compound is also used to study the oxidative metabolism of other structurally related nitrogen-containing drugs.
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| ln Vivo |
Benzydamine N-Oxide does not exhibit in vivo biological activity because it is an inactive metabolite. After oral, topical, or parenteral administration of benzydamine, a fraction is metabolized to benzydamine N-Oxide by hepatic CYP3A4 and FMOs. The N-oxide metabolite is then further metabolized or excreted in the urine. Measuring the ratio of benzydamine to its N-oxide in plasma or urine provides insights into the activity of drug-metabolizing enzymes (e.g., CYP3A4 activity). As an internal standard, the compound can be administered in tracer amounts (if isotopically labeled) to correct for recovery and matrix effects in analytical studies, but it is not intended to produce a biological effect.
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| Enzyme Assay |
A typical non-cellular protocol for using Benzydamine N-Oxide as an internal standard involves its incorporation into LC-MS/MS sample preparation. A stock solution of the internal standard (1 mg/mL) is prepared in methanol. For plasma samples, 50 microL of plasma is transferred to a microcentrifuge tube. Then, 10 microL of internal standard solution (diluted to 1 microg/mL) is added to achieve a final concentration of 100-200 ng/mL. Proteins are precipitated with 150 microL of acetonitrile containing 0.1% formic acid, vortexed, and centrifuged at 14,000 rpm for 10 minutes at 4degC. The supernatant (150 microL) is transferred to an autosampler vial and mixed with 150 microL of water. The sample (5-10 microL) is injected onto an LC-MS/MS system operated in positive ion mode with MRM for the transitions: benzydamine and Benzydamine N-Oxide. The analyte-to-internal standard peak area ratio is used for quantification against a calibration curve.
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| Cell Assay |
An in vitro cellular protocol for using Benzydamine N-Oxide as an analytical standard involves quantifying benzydamine in cultured cells. Primary human hepatocytes or HepG2 cells are seeded in 6-well plates at 1×10⁶ cells/well and cultured for 24 hours. Cells are treated with 10 microM benzydamine (non-labeled) for 0.5-4 hours. The culture medium is collected. Cells are washed with PBS and harvested by scraping. The cell pellet is resuspended in 200 microL of PBS. Benzydamine N-Oxide (10 microL of 1 microg/mL) is added to both medium and cell lysate. Proteins are precipitated with 800 microL of acetonitrile. After centrifugation, the supernatant is analyzed by LC-MS/MS. The concentration of benzydamine in the medium and cells is quantified using a calibration curve. The internal standard corrects for extraction efficiency and matrix effects.
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| Animal Protocol |
An in vivo animal protocol for using Benzydamine N-Oxide as an internal standard involves the quantification of benzydamine and its metabolites. Male Sprague-Dawley rats (200-250 g) are administered benzydamine via oral gavage (10-20 mg/kg) or intravenous injection (5 mg/kg). Blood samples (200 microL) are collected via the tail vein into EDTA-coated tubes at various time points (0, 0.5, 1, 2, 4, 6, 8, 12, 24 hours) post-dose. Plasma is separated by centrifugation. For extraction, 50 microL of plasma is mixed with 10 microL of Benzydamine N-Oxide internal standard (1 microg/mL) and 150 microL of acetonitrile. After centrifugation, the supernatant is analyzed by LC-MS/MS. The concentrations of benzydamine and benzydamine N-oxide are quantified using a calibration curve. The internal standard corrects for variations in sample preparation and matrix effects.
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| ADME/Pharmacokinetics |
Benzydamine N-Oxide, as an internal standard, is not characterized by typical pharmacokinetic parameters. The parent drug benzydamine has the following PK properties: After oral administration, benzydamine is rapidly absorbed with a Tmax of 1-2 hours, an elimination half-life (t½) of approximately 4-6 hours, and an oral bioavailability of about 50% due to first-pass metabolism. Benzydamine N-Oxide is formed as a major metabolite and reaches peak concentrations slightly later than the parent compound. The N-oxide metabolite is eliminated in the urine. The deuterated internal standard, when spiked into samples, does not contribute to the in vivo PK profile.
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| Toxicity/Toxicokinetics |
Toxicity data specific to Benzydamine N-Oxide are not available, as it is a metabolite and analytical standard. The parent compound benzydamine has a favorable safety profile when used topically, but oral administration can cause gastrointestinal discomfort, nausea, and dizziness. Benzydamine N-Oxide is less active than the parent drug and is not considered toxic. Standard laboratory safety precautions should be followed when handling the compound, including the use of gloves and safety glasses. The compound should be stored at -20degC, protected from light and moisture. It is for research use only and should not be used in humans for therapeutic or diagnostic purposes.
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| References | |
| Additional Infomation |
Benzydamine N-oxide is the parent compound; its structural formula is shown in the first source; it is a metabolite of benzalkonium chloride.
Benzydamine N-Oxide (CAS 36504-71-9) is a stable, analytically useful internal standard for the quantification of the non-steroidal anti-inflammatory drug benzydamine. It is the major metabolite of benzydamine, formed primarily by CYP3A4 and FMOs. The compound is valuable for LC-MS/MS method development for pharmacokinetic studies and for investigating drug-drug interactions. It is also used as a marker of oxidative metabolism. Benzydamine N-Oxide is not pharmacologically active and has no significant anti-inflammatory or analgesic properties. The molecular formula is C19H23N3O3, and the molecular weight is 341.41. It is intended for research use only. |
| Molecular Formula |
C19H23N3O2
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|---|---|
| Molecular Weight |
325.40
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| Exact Mass |
122.061
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| CAS # |
36504-71-9
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| PubChem CID |
65465
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| Appearance |
Colorless to light yellow ointment
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| LogP |
2.167
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| Hydrogen Bond Donor Count |
0
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| Hydrogen Bond Acceptor Count |
3
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| Rotatable Bond Count |
7
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| Heavy Atom Count |
24
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| Complexity |
385
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| Defined Atom Stereocenter Count |
0
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| SMILES |
C[N+](CCCOC1=NN(CC2C=CC=CC=2)C2=CC=CC=C12)(C)[O-]
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| InChi Key |
VHKKEFPZHPEYJK-UHFFFAOYSA-N
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| InChi Code |
InChI=1S/C19H23N3O2/c1-22(2,23)13-8-14-24-19-17-11-6-7-12-18(17)21(20-19)15-16-9-4-3-5-10-16/h3-7,9-12H,8,13-15H2,1-2H3
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| Chemical Name |
3-(1-benzylindazol-3-yl)oxy-N,N-dimethylpropan-1-amine oxide
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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: 50 mg/mL (153.66 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (7.68 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL. Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution. Solubility in Formulation 2: ≥ 2.5 mg/mL (7.68 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution. For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly. Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (7.68 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 3.0731 mL | 15.3657 mL | 30.7314 mL | |
| 5 mM | 0.6146 mL | 3.0731 mL | 6.1463 mL | |
| 10 mM | 0.3073 mL | 1.5366 mL | 3.0731 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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