| Size | Price | Stock | Qty |
|---|---|---|---|
| 5mg |
|
||
| Other Sizes |
| Targets |
Benzoate-d5 sodium is a stable isotope-labeled internal standard. Its unlabeled parent, sodium benzoate, has two primary pharmacological targets and mechanisms of action. First, as an antimicrobial preservative, it acts by inhibiting the growth of yeasts, bacteria, and molds in acidic environments (pH 2.5-4.0). The undissociated benzoic acid (C₆H₅COOH) enters the microbial cell, causing intracellular acidification, disruption of the proton motive force, and inhibition of the metabolism of key enzymes (e.g., alpha-ketoglutarate dehydrogenase). Second, as a drug for the treatment of urea cycle disorders (hyperammonemia), sodium benzoate acts as a nitrogen scavenger. It conjugates with the amino acid glycine in the liver (via benzoyl-CoA synthetase and benzoyl-CoA:glycine N-acyltransferase) to form hippuric acid (benzoylglycine). This reaction consumes nitrogen (in the form of glycine) and provides an alternative pathway for the excretion of waste nitrogen in the urine (as hippurate), thereby lowering plasma ammonia levels. The deuterated version (Benzoate-d5 sodium) is used as a tracer to study these mechanisms.
|
|---|---|
| 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 Benzoate-d5 sodium is presumed to be identical to its unlabeled parent, sodium benzoate. In antimicrobial susceptibility testing (minimum inhibitory concentration, MIC), sodium benzoate inhibits the growth of fungi (e.g., Aspergillus niger, Candida albicans) at concentrations of 1-5 mg/mL (pH 3.0). It is less effective against bacteria. In metabolic studies in primary rat hepatocytes, sodium benzoate (0.1-2 mM) rapidly conjugates with glycine to form hippuric acid. The rate of hippuric acid formation is dependent on the availability of glycine and the activity of the enzymes benzoyl-CoA synthetase and glycine N-acyltransferase (GLYAT). In uremic rats and humans, the conjugation capacity for benzoate is reduced. In cell-free assays, benzoate (10-100 mM) has been shown to inhibit various enzymes, including alpha-ketoglutarate dehydrogenase (IC₅0 ~ 20 mM) and other TCA cycle enzymes. The labeled Benzoate-d5 sodium is used as an internal standard to accurately quantify sodium benzoate and hippuric acid in these in vitro systems by LC-MS. |
| ln Vivo |
The in vivo activity of Benzoate-d5 sodium is not directly evaluated; it is used as an internal standard. Its unlabeled parent, sodium benzoate, is an effective antimicrobial agent in food preservation and an established drug for treating hyperammonemia. In patients with urea cycle disorders (e.g., ornithine transcarbamylase deficiency, OTC deficiency), intravenous or oral administration of sodium benzoate (250-500 mg/kg/day) reduces plasma ammonia levels by providing an alternative pathway for waste nitrogen excretion (as hippuric acid). In rats, oral administration of sodium benzoate (200-1,000 mg/kg) results in a dose-dependent increase in hippuric acid excretion in urine (within 2-4 hours). Up to 70-80% of an oral dose of sodium benzoate is excreted in urine as hippuric acid within 6-12 hours. In humans, the half-life of sodium benzoate is approximately 1-2 hours (for the disappearance of benzoate from the blood). Benzoate is rapidly converted to hippurate, which is then eliminated by the kidneys (glomerular filtration and active secretion). The labeled Benzoate-d5 sodium is used as an internal standard to accurately quantify benzoate and hippurate in plasma and urine samples in clinical and preclinical PK studies.
|
| Enzyme Assay |
A generic non-cell-based assay for Benzoate-d5 sodium involves its use as an internal standard in an LC-MS/MS method for quantifying sodium benzoate and hippuric acid in human plasma. Prepare a standard stock solution of unlabeled sodium benzoate and hippuric acid in water (1 mg/mL each). Prepare a separate stock solution of the internal standard Benzoate-d5 sodium at the same concentration. Prepare calibration standards by spiking the unlabeled analytes into a blank matrix (e.g., charcoal-stripped human plasma) to achieve concentrations ranging from 0.1 to 200 ug/mL for benzoate and 0.5 to 500 ug/mL for hippurate. Add a fixed concentration of the internal standard (e.g., 10 ug/mL) to each calibration standard. For sample preparation, add 200 uL of acetonitrile to 50 uL of plasma to precipitate proteins. Vortex and centrifuge at 12,000g for 10 minutes. Transfer the supernatant to an autosampler vial. Analyze by LC-MS/MS in negative ion mode. Monitor the mass transitions: m/z 121 → 77 for benzoate (loss of CO2), m/z 178 → 134 for hippuric acid (loss of COOH), m/z 126 → 82 for the benzoate-d5 internal standard (due to +5 Da shift), and m/z 183 → 139 for the hippuric acid-d5 (if directly labeled). Construct separate calibration curves for benzoate and hippurate by plotting the peak area ratio (analyte/IS) vs. the nominal concentration.
|
| Cell Assay |
A standard in vitro cell-based protocol for Benzoate-d5 sodium involves the assessment of benzoate conjugation capacity in primary human hepatocytes. Isolate primary human hepatocytes from donor liver tissue or purchase commercially (e.g., from Lonza). Culture the hepatocytes in Williams‘ Medium E supplemented with Matrigel, in 12-well plates at 5×10⁵ cells/well. After 24 hours of attachment, treat the cells with 0.5 mM unlabeled sodium benzoate (dissolved in medium) for 0, 1, 2, 4, 6, 8, 12, and 24 hours. At each time point, collect the culture medium and cells separately. To the medium samples, add a fixed amount of Benzoate-d5 sodium as an internal standard. For the cell lysates, homogenize the cells in 500 uL of PBS, extract with acetonitrile, and add the internal standard. Analyze by LC-MS/MS to quantify the depletion of benzoate from the medium and the production of hippuric acid. Calculate the intrinsic clearance (CL_int) for benzoate metabolism in the hepatocytes. This protocol can be used to assess the effect of hepatotoxins, genetic polymorphisms (in GLYAT), or co-administered drugs on the rate of benzoylglycine conjugation.
|
| Animal Protocol |
A typical in vivo animal protocol for Benzoate-d5 sodium involves a pharmacokinetic (PK) study in rats. Use male Sprague-Dawley rats (250-300 g, n = 5-6 per group). Administer a single oral dose of unlabeled sodium benzoate (200 mg/kg) in water by gavage. Collect blood samples via tail vein at various time points (0, 15, 30, 60, 90, 120, 180, 240, 360, 480 minutes) into heparinized tubes. Centrifuge immediately to obtain plasma. Also collect 24-hour urine samples using metabolic cages. For bioanalysis, spike 50 uL of plasma or 20 uL of urine (diluted 1:10 in water) with a fixed amount of Benzoate-d5 sodium as internal standard. Precipitate proteins with acetonitrile, centrifuge, and analyze the supernatant by LC-MS/MS. Quantify sodium benzoate and hippuric acid (the metabolite). Calculate the PK parameters (Cmax, Tmax, AUC, t½, CL) for both parent and metabolite. In the urine, determine the total amount of hippuric acid excreted over 24 hours and calculate the percentage of the administered dose recovered as the glycine conjugate. This protocol is used in preclinical research to study the influence of disease states (e.g., chronic kidney disease, liver cirrhosis) or genetic differences on the metabolism and excretion of benzoate.
|
| ADME/Pharmacokinetics |
Benzoate-d5 sodium is an analytical internal standard. Its unlabeled parent, sodium benzoate, has a well-defined pharmacokinetic profile. In humans, following oral administration of sodium benzoate (up to 5 grams), benzoate is rapidly absorbed from the small intestine (Tmax ~ 30-60 minutes). It is then taken up by the liver, where it is conjugated with glycine to form hippuric acid in a two-step enzymatic process. The capacity for hippuric acid synthesis is limited (a phenomenon known as the “glycine conjugation bottleneck”), and doses above 5-10 grams can exceed this capacity, leading to benzoate accumulation and toxicity. The elimination half-life of benzoate is approximately 1-2 hours in humans, while that of hippuric acid is about 2-4 hours. The volume of distribution of benzoate is small (~0.1-0.2 L/kg), as it is largely confined to the extracellular fluid. The major route of excretion is urinary, with hippuric acid accounting for >70% of the dose within 12 hours. The labeled Benzoate-d5 sodium is essential for accurate quantification of benzoate and hippurate in clinical pharmacokinetic and pharmacodynamic studies, especially when measuring low concentrations in tissues or studying drug interactions (e.g., with probenecid, which inhibits hippurate secretion).
|
| Toxicity/Toxicokinetics |
Benzoate-d5 sodium is a research-grade stable isotope-labeled compound, not a pharmaceutical drug. Its unlabeled parent, sodium benzoate, is generally recognized as safe (GRAS) as a food preservative at concentrations up to 0.1% (1,000 ppm) in foods and beverages. The oral LD₅0 in rats is 3,000-4,000 mg/kg, indicating low acute toxicity. However, high doses of sodium benzoate (especially in combination with ascorbic acid, vitamin C) can form benzene (a known human carcinogen) in beverages under certain conditions (high heat and light). In the context of medical use (for hyperammonemia), high doses (250-500 mg/kg/day) are used, which can cause side effects such as nausea, vomiting, hypokalemia, and, rarely, neurological symptoms (including seizures) due to benzoate-induced glycine depletion. Occupational exposure to sodium benzoate dust may cause mild eye and skin irritation. For laboratory handling of Benzoate-d5 sodium, standard chemical safety precautions (gloves, lab coat, goggles) are sufficient. The compound is stable and should be stored at room temperature or 2-8degC in a tightly sealed container, protected from moisture. It is for research use only and not for human consumption.
|
| References | |
| Additional Infomation |
Benzoate-d5 sodium (sodium benzoate-d5) is the stable isotope-labeled version of the widely used food preservative and pharmaceutical excipient sodium benzoate (C₆H₅COONa, E211). The compound contains five deuterium atoms on the benzene ring, providing a mass shift of +5 Da. It is intended for research use as an internal standard for the accurate quantification of sodium benzoate and its major glycine conjugate metabolite, hippuric acid, in biological samples (plasma, urine, tissues) by LC-MS/MS or GC-MS. Sodium benzoate has several important applications: (1) as an antimicrobial preservative in acidic foods and beverages; (2) as a drug for the treatment of hyperammonemia in patients with urea cycle disorders (as a nitrogen scavenger); and (3) as an excipient in pharmaceutical formulations (e.g., as a buffer or preservative in liquid medicines). The labeled standard is an essential analytical tool for pharmacokinetic studies, food safety analysis, and clinical monitoring of benzoate therapy. For research use only, not for human consumption.
|
| Molecular Formula |
C7D5NAO2
|
|---|---|
| Molecular Weight |
149.13
|
| Exact Mass |
149.05
|
| CAS # |
62790-26-5
|
| Related CAS # |
Sodium benzoate;532-32-1
|
| PubChem CID |
23671533
|
| Appearance |
White to off-white solid powder
|
| Melting Point |
>300ºC(lit.)
|
| Flash Point |
121ºC
|
| LogP |
0.05
|
| Hydrogen Bond Donor Count |
0
|
| Hydrogen Bond Acceptor Count |
2
|
| Rotatable Bond Count |
1
|
| Heavy Atom Count |
10
|
| Complexity |
108
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
[2H]C1=C(C(=C(C(=C1[2H])[2H])C(=O)[O-])[2H])[2H].[Na+]
|
| InChi Key |
WXMKPNITSTVMEF-GWVWGMRQSA-M
|
| InChi Code |
InChI=1S/C7H6O2.Na/c8-7(9)6-4-2-1-3-5-6;/h1-5H,(H,8,9);/q;+1/p-1/i1D,2D,3D,4D,5D;
|
| Chemical Name |
sodium;2,3,4,5,6-pentadeuteriobenzoate
|
| 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 Note: Please store this product in a sealed and protected environment, avoid exposure to moisture. |
| 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 | 6.7056 mL | 33.5278 mL | 67.0556 mL | |
| 5 mM | 1.3411 mL | 6.7056 mL | 13.4111 mL | |
| 10 mM | 0.6706 mL | 3.3528 mL | 6.7056 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
