| Size | Price | Stock | Qty |
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| 5mg |
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| Targets |
Phosphate dibasic-d1 sodium is a stable isotope-labeled internal standard. Its unlabeled parent, disodium hydrogen phosphate (Na2HPO4), is an inorganic salt, not a drug with a specific pharmacological target. In the body, the phosphate ion (HPO42-/H2PO4-) is an essential nutrient and a key component of numerous biological molecules (e.g., ATP, DNA, RNA, phospholipids, and hydroxyapatite in bones and teeth). Phosphate also plays a critical role in cellular signaling (protein phosphorylation) and acid-base buffering (in blood, the phosphate buffer system is a minor but important buffer). In pharmacology, phosphate salts are used as saline laxatives (to cleanse the bowel before colonoscopy) and as urinary acidifiers. They are also used as excipients (buffers, stabilizers) in drug formulations. The labeled Phosphate dibasic-d1 sodium is used as a tracer to study phosphate absorption, distribution, and excretion, and as an internal standard for the quantification of phosphate in various matrices (e.g., blood, urine, soil). It has no direct biological activity of its own at the concentrations used for analysis.
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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 Phosphate dibasic-d1 sodium is not characterized, as it is used as an analytical standard. Its unlabeled parent, phosphate ion (HPO42-), is an essential component of cell culture media. In cell culture, phosphate is required at concentrations of 0.5-2 mM for optimal cell growth, proliferation, and ATP production. In the presence of calcium, phosphate can form calcium phosphate precipitates (hydroxyapatite-like crystals), which can influence osteoblast differentiation. In osteoblast cell lines (MC3T3-E1), phosphate (2-10 mM) is known to induce mineralization (bone formation) in the presence of beta-glycerophosphate. In kidney cell lines (OK cells, a model for proximal tubule), phosphate uptake occurs via the sodium-dependent phosphate cotransporters (NaPi-IIa, NaPi-IIc). The labeled compound (Phosphate dibasic-d1 sodium) can be used as a tracer to measure the rate of phosphate uptake into cells (e.g., by adding ¹⁸O-labeled phosphate or D-labeled phosphate and analyzing by mass spectrometry). It is also used as an internal standard to accurately quantify the total phosphate concentration in cell culture media and cell lysates. |
| ln Vivo |
The in vivo activity of Phosphate dibasic-d1 sodium is not evaluated; it is used as a tracer and internal standard. In humans and animals, phosphate is an essential nutrient. The unlabeled disodium hydrogen phosphate, when administered orally (as a saline laxative), acts osmotically in the small intestine and colon, drawing water into the bowel lumen and inducing peristalsis (catharsis). The oral laxative dose is approximately 2-4 grams. Intravenous phosphate (as sodium or potassium phosphate) is used to treat hypophosphatemia (low serum phosphate). Phosphate is also a major component of bones and teeth, and it plays a role in pH buffering (phosphate buffer system). The labeled Phosphate dibasic-d1 sodium can be administered as a tracer (via oral gavage or intravenous injection) in animal studies to investigate the kinetics of phosphate absorption from the gut, its distribution into bone and soft tissues, and its renal excretion. By measuring the ratio of labeled to unlabeled phosphate in blood, urine, and tissues by LC-MS, researchers can calculate parameters such as fractional absorption and metabolic flux.
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| Enzyme Assay |
A generic non-cell-based assay for Phosphate dibasic-d1 sodium involves its use as an internal standard in an LC-MS method for quantifying inorganic phosphate in urine or plasma. Prepare a calibration curve using a standard solution of unlabeled disodium hydrogen phosphate in water (0.1-10 mM). Add a fixed concentration of Phosphate dibasic-d1 sodium (e.g., 0.5 mM) as internal standard to each calibration standard. For sample preparation, dilute biological fluids (e.g., plasma or urine) 10-100 fold in water. Precipitate proteins with acetonitrile (if needed) or simply filter. For phosphate analysis by LC-MS, a HILIC (hydrophilic interaction chromatography) column with an ammonium carbonate/acetonitrile mobile phase is commonly used to retain the highly polar phosphate ion. Analyze by LC-MS/MS in negative ion mode. Monitor the mass transitions: m/z 97 → 79 (loss of H2O) for H2PO4-/HPO42-, and m/z 98 → 80 for the deuterated phosphate (D-phosphate). Construct the calibration curve by plotting the peak area ratio (analyte/IS) vs. the nominal concentration. This method is used for the quantification of inorganic phosphate in clinical chemistry and environmental analysis.
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| Cell Assay |
A standard in vitro cell-based protocol for phosphate studies uses the OK cell line (opossum kidney cells) to measure sodium-dependent phosphate uptake. Culture OK cells in DMEM/F12 medium supplemented with 10% FBS, 1% penicillin/streptomycin, and 2 mM L-glutamine at 37degC in a 5% CO2 incubator. Seed cells in 24-well plates at 2×10⁵ cells/well and allow to grow for 48-72 hours until confluent. On the day of the assay, wash the cells twice with pre-warmed uptake buffer (140 mM NaCl, 5 mM KCl, 1 mM CaCl2, 1 mM MgSO4, 10 mM HEPES, pH 7.4). Pre-incubate cells with parathyroid hormone (PTH, 10 nM) or vehicle for 1 hour to downregulate NaPi-IIa expression (negative control). For the uptake assay, add 250 uL of uptake buffer containing 100 uM unlabeled KH2PO4 (or NaH2PO4) and 0.5 uCi/mL of [32P]-orthophosphate (or use stable isotope-labeled phosphate like Phosphate dibasic-d1 sodium as a tracer) for 5 minutes at 37degC. Terminate the uptake by washing the cells three times with ice-cold stop buffer (same as uptake buffer but with 1 mM HgCl2 to inhibit transporters). Lyse the cells in 0.1 N NaOH. Measure the radioactivity in the lysate using a scintillation counter (if using 32P). Alternatively, use the deuterium-labeled phosphate as a tracer and quantify the phosphate content in the lysate by LC-MS. Calculate the phosphate uptake rate (nmol/mg protein/min). This protocol is used to study the regulation of phosphate transport by hormones (PTH, FGF23) and by phosphate itself.
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| Animal Protocol |
A typical in vivo animal protocol for Phosphate dibasic-d1 sodium involves a pharmacokinetic study of phosphate absorption and distribution. Use male C57BL/6J mice (8-10 weeks old, n = 4-5 per time point). Fasting the mice overnight (12-14 hours) with free access to water. Administer a single oral dose of unlabeled disodium hydrogen phosphate (50 mg/kg, equivalent to ~25 mg/kg of phosphate) in water, spiked with a tracer dose of Phosphate dibasic-d1 sodium (1 mg/kg), via oral gavage. Collect blood samples via tail vein at various time points (0, 15, 30, 60, 120, 240, 360 minutes) into heparinized tubes. Immediately centrifuge to obtain plasma. Also collect 24-hour urine samples using metabolic cages. Euthanize the mice at the terminal time point (4 hours) and collect liver, kidney, and bone (femur) tissues. For bioanalysis, deproteinize plasma (10 uL) and urine (50 uL) with acetonitrile. For tissues, homogenize in water and then deproteinize. Analyze the samples by LC-MS (HILIC-MS) to quantify the concentration of total phosphate and the labeled Phosphate dibasic-d1 sodium. Calculate the time course of the labeled phosphate in plasma (AUC), its urinary excretion (fraction of dose), and its incorporation into bone. This protocol can be used to study the effects of genetic manipulations (e.g., knockout of NaPi-IIa or FGF23) on phosphate handling in vivo.
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| ADME/Pharmacokinetics |
Phosphate dibasic-d1 sodium is an analytical internal standard, not a drug. Its unlabeled parent, disodium hydrogen phosphate, is generally recognized as safe (GRAS) as a food additive (E339(ii)) and is widely used as a buffering agent and emulsifier in processed foods and as a pH adjuster in pharmaceutical formulations. The oral LD₅0 of disodium hydrogen phosphate in rats is >2,000 mg/kg, indicating low acute toxicity. The primary toxicological concerns for phosphate salts are related to electrolyte imbalances when taken in excessive amounts (e.g., as a laxative). Hyperphosphatemia (elevated serum phosphate) can occur in patients with chronic kidney disease (CKD) who are unable to excrete phosphate, leading to secondary hyperparathyroidism and vascular calcification. For laboratory handling of Phosphate dibasic-d1 sodium, standard safety precautions (gloves, lab coat, safety goggles) are sufficient. The compound is stable, non-flammable, and non-explosive. Store at room temperature in a tightly sealed container, protected from moisture (it may be hygroscopic). For research use only, not for human diagnostic or therapeutic applications.
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| References | |
| Additional Infomation |
Phosphate dibasic-d1 sodium (disodium hydrogen phosphate-d1) is the stable isotope-labeled version of disodium hydrogen phosphate (Na2HPO4), an inorganic phosphate salt widely used as a buffer in biochemistry, molecular biology, and pharmaceutical formulations. The compound contains a single deuterium atom bound to the phosphate group (P-OD), giving it a mass shift of +1 Da. It is intended for research use as an internal standard for the accurate quantification of inorganic phosphate in biological samples (urine, plasma, cell lysates) and environmental samples (soil, water) by LC-MS. Inorganic phosphate is an essential nutrient and a key component of ATP, nucleic acids, and bone mineral (hydroxyapatite). It also plays a critical role in cellular signaling via protein phosphorylation and in acid-base buffering. The labeled compound is also used as a tracer in metabolic studies to investigate phosphate absorption, distribution, excretion, and bone mineralization (calcium phosphate deposition). For research use only, not for diagnostic or therapeutic applications.
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| Molecular Formula |
DH2O4P.2NA
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|---|---|
| Molecular Weight |
142.96500
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| Exact Mass |
142.947
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| CAS # |
107632-22-4
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| Related CAS # |
Disodium phosphate;7558-79-4
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| PubChem CID |
102602173
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| Appearance |
White to off-white solid powder
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| LogP |
0
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
0
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| Heavy Atom Count |
7
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| Complexity |
46.5
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| Defined Atom Stereocenter Count |
0
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| SMILES |
[Na+].[Na+].[2H]OP([O-])(=O)[O-]
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| InChi Key |
BNIILDVGGAEEIG-DYCDLGHISA-L
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| InChi Code |
InChI=1S/2Na.H3O4P/c;;1-5(2,3)4/h;;(H3,1,2,3,4)/q2*+1;/p-2/i/hD
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| Chemical Name |
disodium;deuterio phosphate
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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: 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)
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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 | 6.9945 mL | 34.9724 mL | 69.9447 mL | |
| 5 mM | 1.3989 mL | 6.9945 mL | 13.9889 mL | |
| 10 mM | 0.6994 mL | 3.4972 mL | 6.9945 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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