| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
Alverine-d5 citrate is a stable isotope-labeled internal standard. The unlabeled parent compound, Alverine citrate, is a selective smooth muscle relaxant that acts as a functional antagonist at serotonin (5-HT) and muscarinic acetylcholine receptors (mAChRs). However, its primary mechanism of action is believed to be the inhibition of voltage-dependent L-type calcium channels (blocking calcium influx) and phosphodiesterase (PDE) in the smooth muscle cells of the gastrointestinal (GI) tract. It also has affinity for sigma-1 receptors and may modulate neurotransmission in the enteric nervous system. By blocking these targets, Alverine directly relaxes the smooth muscle of the colon, small intestine, and uterus, relieving spasms, pain, and discomfort without affecting normal gut motility. Alverine-d5 citrate mimics these targets for analytical tracing, not for pharmacologic effect.
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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].
The in vitro biological activity of Alverine-d5 citrate is not characterized, as it is an internal standard. However, its unlabeled parent, Alverine citrate, shows specific activity in cell-based assays. In isolated smooth muscle preparations (e.g., guinea pig ileum or rat colon), Alverine (1-100 uM) concentration-dependently inhibits contractions induced by a variety of spasmogens, including acetylcholine, histamine, serotonin, and barium chloride. In cultured colonic smooth muscle cells, Alverine (10-50 uM) has been shown to block the intracellular calcium rise induced by agonists like carbachol, as measured by fluorescent calcium indicators (e.g., Fura-2). In non-cell-based assays, Alverine binds to the 5-HT1A receptor with moderate affinity (Ki = 0.5 uM). It also inhibits type-4 phosphodiesterase (PDE4) with an IC₅0 in the low micromolar range. Alverine-d5 citrate is used as an internal standard to accurately quantify the drug in the organ bath or culture media in these experiments. |
| ln Vivo |
Alverine-d5 citrate is not used for efficacy studies in animals, but its unlabeled parent is well-studied. In vivo, Alverine citrate exhibits antispasmodic activity in various animal models. In anesthetized dogs and rats, intravenous administration of Alverine (1-5 mg/kg) markedly reduces the tone and spontaneous motility of the colon and jejunum without significantly affecting blood pressure or heart rate. In a mouse model of diarrhea (e.g., castor oil-induced diarrhea), oral administration of Alverine (10-50 mg/kg) significantly reduced the number of fecal pellets and fluid content, indicating its ability to reduce gastrointestinal hypermotility. Unlike atropine, Alverine does not cause significant anticholinergic side effects like dry mouth or blurred vision at therapeutic doses. Alverine-d5 citrate is used as an internal standard in LC-MS for the precise quantification of the drug in plasma and tissues in these animal models.
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| Enzyme Assay |
Alverine-d5 citrate is an analytical internal standard; therefore, a generic non-cell-based assay involves its use in an LC-MS/MS method for quantification of Alverine. Prepare a standard stock solution of unlabeled Alverine citrate in methanol (1 mg/mL). Prepare a separate stock solution of the internal standard (Alverine-d5 citrate) at the same concentration. Prepare calibration standards by spiking the unlabeled analyte into a blank matrix (e.g., blank human plasma) to achieve concentrations ranging from 0.1 to 500 ng/mL. Add a fixed concentration of the internal standard (e.g., 25 ng/mL) to each calibration standard. Also prepare blank and double-blank samples. For sample preparation, add 200 uL of acetonitrile to 100 uL of plasma for protein precipitation. Vortex and centrifuge at 14,000g for 5 minutes. Transfer the supernatant to an autosampler vial. Analyze by LC-MS/MS in positive ion mode. Monitor the mass transitions: m/z 282.3 → 224.2 for Alverine, and m/z 287.3 → 229.2 for Alverine-d5. Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration. This curve is used to determine the concentration of Alverine in unknown samples.
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| Cell Assay |
A standard in vitro cell-based assay for the unlabeled Alverine uses isolated rat colonic smooth muscle cells. Prepare primary smooth muscle cells from the colon of male Sprague-Dawley rats using an enzymatic digestion method (collagenase). Culture the isolated cells in DMEM supplemented with 10% FBS until they reach 80% confluence. Seed the cells in a 96-well plate and load them with the calcium-sensitive fluorescent dye Fluo-4 AM (2 uM) for 30 minutes. Wash the cells twice with Hanks‘ Balanced Salt Solution (HBSS). Measure fluorescence (Ex/Em = 494/506 nm) using a fluorescence plate reader. Add increasing concentrations of unlabeled Alverine citrate (1 nM to 100 uM) and measure baseline fluorescence. Then, stimulate the cells with 10 uM acetylcholine (ACh) or 60 mM KCl to induce calcium influx. Record the peak fluorescence (calcium transient). Calculate the percentage inhibition of the calcium signal. Determine the half-maximal inhibitory concentration (IC₅0) from the dose-response curve. Use Alverine-d5 citrate as an internal standard in an LC-MS analysis of the HBSS to accurately confirm the drug concentrations in the culture medium.
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| Animal Protocol |
A typical in vivo experiment for Alverine-d5 citrate is a pharmacokinetic (PK) study in beagle dogs. Use male beagle dogs (10-12 kg, n=4-6). Administer a single oral dose of unlabeled Alverine citrate (50 mg) via gavage. Collect blood samples (approx. 2 mL) from the jugular vein at various time points (0, 0.25, 0.5, 0.75, 1, 1.5, 2, 3, 4, 6, 8, 12, 24 h). Centrifuge the blood samples to obtain plasma and store at -80degC until analysis. For bioanalysis, spike an aliquot (100 uL) of each plasma sample with a fixed amount of the internal standard (Alverine-d5 citrate). Perform protein precipitation by adding 300 uL of acetonitrile, vortex, and centrifuge. Inject the supernatant into the LC-MS/MS system. Quantify the concentration of Alverine in each sample using the calibration curve prepared in the same matrix. Calculate the PK parameters (Cmax, Tmax, AUC(0-t), AUC(0-∞), t½, and oral clearance (CL/F)) using non-compartmental analysis with software like Phoenix WinNonlin. This protocol is essential for determining the oral bioavailability and elimination profile of Alverine.
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| ADME/Pharmacokinetics |
Alverine-d5 citrate is an internal standard, so its PK is identical to its non-labeled parent. Alverine citrate is a spasmolytic drug. Following oral administration, Alverine is rapidly absorbed, reaching peak plasma concentrations (Cmax) within 1-2 hours. However, it undergoes extensive first-pass metabolism in the liver via oxidative deamination and O-dealkylation, resulting in very low oral bioavailability (less than 1% for the parent drug in some species). Its primary active metabolite is 4-hydroxy Alverine, which is thought to contribute significantly to its therapeutic effect. The elimination half-life of Alverine is short, approximately 0.5-1 hour. The drug is primarily excreted as metabolites in the urine. The labeled Alverine-d5 is critical for accurately measuring the very low plasma concentrations of the parent drug, as well as its metabolites, thereby enabling precise PK profiling in research settings.
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| Toxicity/Toxicokinetics |
Alverine-d5 citrate is a research-grade chemical and is not for human use. The toxicity of its unlabeled parent, Alverine citrate, is well understood from its clinical use. Alverine is generally considered safe and well-tolerated. The most common side effects in humans are mild and include nausea, headache, dizziness, and gastrointestinal disturbances. Rare cases of allergic skin reactions and hepatitis have been reported. The oral LD₅0 of Alverine citrate in rats is >1000 mg/kg, indicating low acute toxicity. In reproductive toxicology studies, it was not teratogenic in rats and rabbits at clinically relevant doses. The compound does not exhibit genotoxic potential. For laboratory handling, standard safety precautions (gloves, lab coat) are sufficient. Alverine-d5 citrate should be stored as a powder at -20degC to maintain stability.
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| References | |
| Additional Infomation |
Alverine-d5 citrate is the stable isotope-labeled (deuterated) version of Alverine citrate, a smooth muscle relaxant used clinically for the treatment of irritable bowel syndrome (IBS) and other functional gastrointestinal disorders characterized by spasm. It is intended for research use only and serves as an internal standard for the accurate quantification of Alverine in biological matrices by LC-MS/MS. Alverine citrate acts directly on the smooth muscle of the gut without the anticholinergic side effects (like dry mouth) typically associated with other antispasmodics. Its exact mechanism of action is believed to involve calcium channel blockade and serotonin receptor antagonism. This deuterated standard is essential for precise pharmacokinetic and bioequivalence studies of Alverine formulations, as it corrects for matrix effects and variable extraction recoveries in mass spectrometry-based assays.
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| Molecular Formula |
C26H30D5NO7
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| Molecular Weight |
478.59
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| Exact Mass |
478.272
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| CAS # |
1215327-00-6
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| PubChem CID |
46780131
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| Appearance |
White to off-white solid powder
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| Hydrogen Bond Donor Count |
4
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
14
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| Heavy Atom Count |
34
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| Complexity |
442
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[2H]C([2H])([2H])C([2H])([2H])N(CCCC1=CC=CC=C1)CCCC2=CC=CC=C2.C(C(=O)O)C(CC(=O)O)(C(=O)O)O
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| InChi Key |
RYHCACJBKCOBTJ-LUIAAVAXSA-N
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| InChi Code |
InChI=1S/C20H27N.C6H8O7/c1-2-21(17-9-15-19-11-5-3-6-12-19)18-10-16-20-13-7-4-8-14-20;7-3(8)1-6(13,5(11)12)2-4(9)10/h3-8,11-14H,2,9-10,15-18H2,1H3;13H,1-2H2,(H,7,8)(H,9,10)(H,11,12)/i1D3,2D2;
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| Chemical Name |
2-hydroxypropane-1,2,3-tricarboxylic acid;N-(1,1,2,2,2-pentadeuterioethyl)-3-phenyl-N-(3-phenylpropyl)propan-1-amine
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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 | 2.0895 mL | 10.4474 mL | 20.8947 mL | |
| 5 mM | 0.4179 mL | 2.0895 mL | 4.1789 mL | |
| 10 mM | 0.2089 mL | 1.0447 mL | 2.0895 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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