| Size | Price | Stock | Qty |
|---|---|---|---|
| 50mg |
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| Other Sizes |
| Targets |
Although no specific target has been identified for this impurity, it is structurally related to Bumetanide, which inhibits the Na+-K+-2Cl- symporter (NKCC2) in the thick ascending limb of the loop of Henle. The impurity contains an additional butyl ester group that likely reduces binding affinity to NKCC2. Its analytical role does not require receptor binding data, and it is not intended for pharmacodynamic studies.
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|---|---|
| ln Vitro |
No quantitative in vitro activity data have been reported. Based on structural analogy to Bumetanide (IC50 ~0.1 uM for NKCC2 inhibition in oocyte expression systems), this impurity is predicted to have substantially lower inhibitory potency due to steric hindrance from the extra butyl chain. It may show <10% of the parent drug's activity, but this has not been experimentally validated.
|
| ln Vivo |
No in vivo activity studies have been conducted. As an impurity present only at trace levels (typically <0.15% in drug substance), it does not contribute to the diuretic or antihypertensive effects of Bumetanide in patients. Animal efficacy models for diuretics (e.g., urine output measurement in rats) are not applicable to this impurity.
|
| Enzyme Assay |
Non-cell structural characterization: ¹H NMR and ¹3C NMR in DMSO-d₆ or CDCl3 (400 MHz) are used to confirm the structure. LC-MS analysis on a C18 column (150 × 4.6 mm, 5 um) with mobile phase A (0.1% formic acid in water) and B (acetonitrile) at 1.0 mL/min, gradient elution: 5→95% B over 20 min. ESI positive mode detects [M+H]+ at m/z 421.2. HPLC-UV at 254 nm determines purity (>95%).
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| Cell Assay |
A general impurity cytotoxicity protocol uses HEK293 or MDCK cells. Cells are seeded in 96-well plates (1×10⁴ cells/well) and treated with 0.1-100 uM impurity for 24 h. MTT assay measures viability. No specific NKCC2 functional assay (e.g., rubidium uptake) has been published because the impurity is not intended for cell-based pharmacological testing.
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| Animal Protocol |
General impurity qualification in Sprague-Dawley rats: oral gavage at doses of 0.5, 2, and 10 mg/kg for 14 consecutive days (n=10/group). Endpoints include body weight, clinical signs, serum biochemistry (BUN, creatinine, electrolytes), and histopathology of kidney and liver. A control group receives vehicle only. This follows ICH Q3A guidelines for qualification of impurities above the identification threshold.
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| ADME/Pharmacokinetics |
No PK data specific to this impurity. For Bumetanide in humans: oral bioavailability ~80%, Tmax 0.5-2 h, plasma protein binding ~95%, t½ ~1-1.5 h. The impurity, being more lipophilic (estimated log P ~4.5 vs. 2.1 for Bumetanide), may have slower clearance, higher protein binding (>98%), and a longer half-life (~3-6 h in rats), but this is hypothetical.
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| Toxicity/Toxicokinetics |
No dedicated toxicity studies. According to ICH M7, the impurity is assessed for genotoxicity using in silico tools (Derek Nexus, Sarah Nexus). Structural alerts for aromatic amines or sulfonamides may be present. A bacterial reverse mutation (Ames) test (TA98, TA100, TA1535, TA1537, WP2uvrA) with and without metabolic activation (S9) is recommended for qualification.
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| Additional Infomation |
This compound is also known as Bumetanide USP Related Compound C. It appears as a white to off-white crystalline powder. Storage: 2-8 degC, protected from light and moisture. Soluble in DMSO, methanol, and ethanol; practically insoluble in water. Used primarily as an impurity standard in ANDA filings for generic Bumetanide products.
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| Molecular Formula |
C21H28N2O5S
|
|---|---|
| Molecular Weight |
420.52
|
| Exact Mass |
420.172
|
| CAS # |
32643-00-8
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| Related CAS # |
Bumetanide impurity 2
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| PubChem CID |
122917
|
| Appearance |
White to off-white solid powder
|
| Hydrogen Bond Donor Count |
2
|
| Rotatable Bond Count |
12
|
| Heavy Atom Count |
29
|
| Complexity |
586
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
CCCCNC1=C(C(=CC(=C1)C(=O)OCCCC)S(=O)(=O)N)OC2=CC=CC=C2
|
| InChi Key |
FUBXAOXPLXHBPG-UHFFFAOYSA-N
|
| InChi Code |
InChI=1S/C21H28N2O5S/c1-3-5-12-23-18-14-16(21(24)27-13-6-4-2)15-19(29(22,25)26)20(18)28-17-10-8-7-9-11-17/h7-11,14-15,23H,3-6,12-13H2,1-2H3,(H2,22,25,26)
|
| Chemical Name |
butyl 3-(butylamino)-4-phenoxy-5-sulfamoylbenzoate
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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 Note: This product requires protection from light (avoid light exposure) during transportation and storage. |
| 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 | 2.3780 mL | 11.8900 mL | 23.7801 mL | |
| 5 mM | 0.4756 mL | 2.3780 mL | 4.7560 mL | |
| 10 mM | 0.2378 mL | 1.1890 mL | 2.3780 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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