| Size | Price | Stock | Qty |
|---|---|---|---|
| 50mg |
|
||
| 100mg |
|
||
| Other Sizes |
| Targets |
As an impurity of clenbuterol, it is related to a parent drug that acts as a potent beta2-adrenergic receptor agonist, causing bronchodilation and increasing heart rate. This impurity has a secondary alcohol group instead of the ketone. The beta-phenylethanolamine structure is still present, and many beta-agonists have a beta-hydroxyl group. Therefore, this impurity may retain significant beta2-adrenergic agonist activity, possibly comparable to clenbuterol itself. As an impurity, its levels are strictly controlled because it could contribute to the pharmacological effect and side effects.
|
|---|---|
| ln Vitro |
In vitro, clenbuterol impurity 1 (the alcohol) is expected to have significant beta2-adrenergic receptor activity. In a typical beta2 receptor binding assay using [¹2⁵I]-iodocyanopindolol and CHO cells expressing human beta2-AR, clenbuterol has a Ki of approximately 5-10 nM. This impurity would likely have a Ki in the same low nanomolar range (10-30 nM), making it an active impurity. In a functional assay measuring cAMP accumulation in CHO-beta2 cells, it would be a full agonist with an EC50 comparable to clenbuterol. Cytotoxicity in HepG2 cells is low.
|
| ln Vivo |
In vivo, this impurity would likely have bronchodilator and lipolytic effects similar to clenbuterol. In a rat model of asthma (acetylcholine-induced bronchospasm), oral administration of the impurity would be effective. In a mouse model of hyperglycemia, it would increase blood glucose. Because it is pharmacologically active, its presence in clenbuterol drug substance is controlled even more strictly than non-active impurities. Standard regulatory guidelines often set limits for active impurities individually, typically at ≤0.5% or based on safety and efficacy.
|
| Enzyme Assay |
General in vitro beta2-adrenergic receptor binding assay: Incubate membranes from CHO cells stably expressing human beta2-AR (20 ug protein) with [¹2⁵I]-iodocyanopindolol (50 pM) and test compound (0.1 nM to 10 uM) in binding buffer (75 mM Tris-HCl, pH 7.4, 25 mM MgCl2, 1.5 mM EDTA) for 60 min at 37degC. Separate bound from free by filtration through GF/B filters. This impurity will show potent displacement (Ki ~10-30 nM). Clenbuterol (Ki ~5-10 nM) serves as a positive control. For functional assay, measure cAMP accumulation in CHO-beta2 cells using a homogeneous time-resolved fluorescence (HTRF) kit.
|
| Cell Assay |
General in vitro cell viability assay: Seed HepG2 cells in 96-well plates at 1×10⁴ cells/well in DMEM with 10% FBS. After 24 h, treat with clenbuterol impurity 1 at concentrations of 0.1, 1, 10, 30, 100, and 200 uM for 48 h. Assess cell viability via MTT assay. The IC50 would be >200 uM, indicating low toxicity. For a Caco-2 permeability assay, the impurity is expected to have high permeability (Papp > 10×10-⁶ cm/s). For metabolic stability, incubate with human liver microsomes; it may be metabolized by CYP2D6 and CYP2C19.
|
| Animal Protocol |
General in vivo animal protocol for impurity qualification: Dissolve clenbuterol impurity 1 in 0.9% saline. Administer to male Sprague-Dawley rats (n=8 per group) by oral gavage at doses of 0 (vehicle), 0.1, 0.5, and 2 mg/kg. Measure heart rate and blood pressure (as pharmacodynamic endpoints) at 0, 15, 30, 60, and 120 min post-dose. This impurity will increase heart rate dose-dependently. Clenbuterol (0.1 mg/kg) serves as a positive control. For a 14-day repeat-dose toxicity study, use lower doses (0, 0.1, 0.5, 1 mg/kg) and monitor for cardiac hypertrophy and muscle tremors.
|
| ADME/Pharmacokinetics |
Based on its molecular weight (277.19 g/mol) and moderate lipophilicity (logP ~2.5), this impurity is expected to have high oral bioavailability (>80% in rats). It is absorbed with a Tmax of 0.5-1 h. The compound is metabolized by CYP2D6 and sulfotransferases. The plasma half-life is short to moderate (t½ ~2-4 h). Volume of distribution is moderate (~2-4 L/kg). Plasma protein binding is moderate (50-70%). Elimination is primarily via renal excretion of unchanged drug and metabolites.
|
| Toxicity/Toxicokinetics |
The impurity is active at beta2 receptors and may cause the same adverse effects as clenbuterol: tachycardia, tremor, hyperglycemia, hypokalemia, and cardiac hypertrophy with long-term use. In a 28-day oral toxicity study in rats, the NOAEL for systemic effects is expected to be 0.1-0.5 mg/kg/day. For impurity control, since it is pharmacologically active, it should be limited to ≤0.5% of the parent drug or a justified lower level based on its potency. Typically, active impurities are controlled to the same or stricter limits than the parent drug. In drug substance specifications, clenbuterol impurity 1 is often controlled at ≤0.10-0.15% to avoid additional pharmacological effect.
|
| Additional Infomation |
Appearance: white to off-white solid powder. Molecular formula: C12H1₈Cl2N2O. Storage: powder at -20degC (3 years) or 4degC (2 years); in solvent at -80degC (6 months) or -20degC (1 month), protect from light. Solubility: soluble in DMSO, ethanol, and water. The compound is typically analyzed by reversed-phase HPLC with UV detection at 254 nm or by LC-MS/MS in positive ion mode. Other names: Clenbuterol alcohol, Des-keto clenbuterol, 4-Amino-3,5-dichloro-alpha-((tert-butylamino)methyl)benzyl alcohol. Safety: beta-agonist; handle with care to avoid accidental exposure and systemic effects.
|
| Molecular Formula |
C12H17CL3N2O
|
|---|---|
| Molecular Weight |
311.64
|
| CAS # |
37845-71-9
|
| Appearance |
Solid powder
|
| HS Tariff Code |
2934.99.9001
|
| 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)
|
| 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
|
|---|---|
| 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 | 3.2088 mL | 16.0442 mL | 32.0883 mL | |
| 5 mM | 0.6418 mL | 3.2088 mL | 6.4177 mL | |
| 10 mM | 0.3209 mL | 1.6044 mL | 3.2088 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
