None (biochemical reagent). Glycerophosphoric acid (also known as glycerophosphate) is a phosphorylated glycerol derivative. It is an endogenous metabolite involved in phospholipid metabolism and energy metabolism. It serves as a precursor for phospholipid synthesis (e.g., phosphatidylcholine, phosphatidylethanolamine) and is also a component of the glycerol phosphate shuttle, which transfers reducing equivalents from cytosolic NADH into mitochondria. As a disodium salt, it provides sodium ions and phosphate groups. In research, beta-glycerophosphate is widely used as a source of inorganic phosphate in cell culture media, especially to induce differentiation and mineralization of osteoblasts, chondrocytes, and mesenchymal stem cells. The alpha and beta isomers differ in the position of the phosphate group on the glycerol backbone. The alpha isomer has the phosphate on the primary carbon, while the beta isomer has the phosphate on the secondary carbon. This product does not target any specific receptor or enzyme in a pharmacological sense; it is a simple chemical reagent.

In vitro, beta-glycerophosphate (a major component of this mixture) is a standard component of osteogenic differentiation media. At concentrations of 5-10 mM, it provides an organic phosphate source that is hydrolyzed by alkaline phosphatase (ALP) to release inorganic phosphate. The released phosphate combines with calcium ions to form calcium phosphate deposits, leading to mineralization of the extracellular matrix, as evidenced by Alizarin Red S staining or von Kossa staining. beta-Glycerophosphate alone does not induce osteoblast differentiation; it is typically used in combination with ascorbic acid (50-100 ug/mL) and dexamethasone (10-100 nM). In chondrocyte cultures, beta-glycerophosphate (5-10 mM) is used to promote cartilage matrix mineralization during hypertrophic differentiation. The alpha and beta mixture may have similar activity, but beta-glycerophosphate is the more common isomer in differentiation media. The compound also serves as a substrate for alkaline phosphatase activity assays; when incubated with ALP, it releases inorganic phosphate, which can be measured colorimetrically. Glycerophosphoric acid can also be used as a standard for metabolomics studies. The disodium salt and hydrate form do not alter its biochemical activity.

Not applicable (biochemical reagent). Glycerophosphoric acid disodium salt hydrate is not intended for in vivo administration as a therapeutic agent. It is a laboratory reagent used in cell culture and biochemical assays. When administered systemically (e.g., intravenously), glycerophosphate can serve as a source of phosphate, but this is not the intended use of the research-grade product. There are no reported studies of its in vivo efficacy in animal disease models, as it is not a drug candidate. The compound is generally recognized as safe (GRAS) as a food additive but is not approved for any medical indication. For in vivo use in animals (e.g., in phosphate replacement therapies), clinical-grade phosphate salts are used, not research-grade glycerophosphoric acid. Therefore, no in vivo activity data are available for this product as a "drug."

Not applicable. Glycerophosphoric acid disodium salt hydrate is a simple organic salt and is not typically used in receptor or enzyme binding assays as a ligand. However, it can serve as a substrate for enzymatic assays such as alkaline phosphatase (ALP). For ALP activity measurement: 96-well plates are set up with assay buffer (100 mM glycine-NaOH, pH 10.4, 1 mM MgCl2). Serial dilutions of glycerophosphoric acid (0.5-20 mM) are added, and purified ALP (0.1-1 U/mL) is added to initiate the reaction. The reaction mixture is incubated at 37degC for 15-30 minutes. The product inorganic phosphate is measured using a malachite green phosphate assay or a commercial kit (e.g., QuantiChrom). Briefly, 50 uL of the reaction supernatant is mixed with 100 uL of malachite green reagent, incubated for 10 min, and absorbance is read at 620 nm. A standard curve using phosphate standards (0.1-5 mM) is used for quantification. The Km and Vmax of ALP for glycerophosphate can be determined using Lineweaver-Burk plots. Alternatively, a fluorogenic substrate (e.g., 4-methylumbelliferyl phosphate) is more sensitive, but glycerophosphoric acid is not suitable for fluorescent detection. For binding to other proteins or enzymes, no specific protocols are available. The compound is not a high-affinity ligand for any known receptor; it is a substrate or metabolite.

For cell culture studies, the primary use of beta-glycerophosphate (as a component of the alpha/beta mixture) is to induce osteogenic differentiation. Mesenchymal stem cells (MSCs), osteoblasts (e.g., MC3T3-E1), or other cell types are seeded in 6-well or 24-well plates at 1-3 × 10^4 cells/cm2 in growth medium (alpha-MEM or DMEM with 10% FBS, 100 U/mL penicillin, 100 ug/mL streptomycin). When cells reach 70-80% confluence (typically day 2-3), the medium is replaced with osteogenic differentiation medium containing: 5-10 mM beta-glycerophosphate (from the alpha/beta mixture), 50-100 ug/mL ascorbic acid, and 10-100 nM dexamethasone (optional). Control cultures receive growth medium without osteogenic supplements. Cells are cultured for 7-28 days, with medium changed every 2-3 days. Mineralization is assessed by Alizarin Red S staining: Cells are fixed with 4% formalin for 15 min, washed with water, and stained with 40 mM Alizarin Red S (pH 4.2) for 20 min at room temperature. After washing, stain is destained with 10% cetylpyridinium chloride for 1 hour and quantified by absorbance at 562 nm. Alternatively, von Kossa staining is used for phosphate deposits. Alkaline phosphatase (ALP) activity is measured by adding 0.1% Triton X-100 to cells, then incubating lysate with p-nitrophenyl phosphate (pNPP) in ALP buffer (100 mM glycine, 1 mM MgCl2, pH 10.4) at 37degC for 30 min; absorbance at 405 nm is measured. For cell viability: the compound is not cytotoxic at 5-10 mM; higher concentrations (50-100 mM) may cause osmotic stress. For cell proliferation assays (MTT), cells are treated with various concentrations (0-50 mM) for 24-72 h. The product is water-soluble; prepare stock solution at 0.5-1 M in water, filter-sterilize, and store at 4degC. The pH of the medium should be adjusted to 7.4 after adding glycerophosphate. For chondrocyte differentiation: Pellet cultures of chondrocytes are maintained in serum-free medium with 5-10 mM beta-glycerophosphate for 14-21 days, and pellets are stained with Alcian blue or Safranin O for glycosaminoglycans. The alpha isomer may have different activities; but the mixture is often used as supplied. All experiments should be performed in triplicate wells.

Not applicable (in vitro reagent). Glycerophosphoric acid disodium salt hydrate is not used as a drug in animal studies; however, beta-glycerophosphate (the active isomer) can be administered to animals in research contexts as a phosphate source or to study the effect of phosphate on mineralization. For a typical in vivo study of heterotopic ossification (ectopic bone formation): C57BL/6 mice are anesthetized, and a mixture of recombinant human bone morphogenetic protein 2 (rhBMP-2, 5 ug) with 10 mM beta-glycerophosphate (from the mixture) in a collagen scaffold is implanted intramuscularly (e.g., in the thigh). After 2-4 weeks, the implant is harvested, fixed, and analyzed by micro-CT and histology (H&E, Alizarin Red S) for mineralized tissue formation. However, this is not a drug administration protocol; the compound is used as a component of an implant. For systemic phosphate supplementation (e.g., in hypophosphatemia models), glycerophosphate is rarely used; inorganic phosphate salts are preferred. Therefore, no detailed in vivo animal experimental protocol for this product as a "drug" is available. The compound is for research use only, not for animal treatment.

Not applicable. Glycerophosphoric acid disodium salt hydrate is not a drug, and no pharmacokinetic data are available for this compound in the context of drug development. As a simple organic phosphate, it is water-soluble, not expected to bind to plasma proteins, and rapidly cleared by the kidneys if administered intravenously. beta-Glycerophosphate is a metabolite and can be hydrolyzed by alkaline phosphatase (ALP) to release glycerol and inorganic phosphate. In plasma, its half-life is likely minutes, as it is quickly hydrolyzed. The alpha and beta isomers may have slightly different rates of hydrolysis. The compound is not intended for use as a therapeutic agent. For research involving cellular uptake, radiolabeled glycerophosphate (e.g., 14C-labeled) could be used to track metabolism, but such studies are not standard. The hydrate and sodium salt forms do not alter its PK. This product is strictly a laboratory reagent.

No specific toxicity data are available for glycerophosphoric acid disodium salt hydrate. beta-Glycerophosphate is generally considered low in toxicity. In vitro, concentrations up to 20-30 mM are tolerated by most cell types for short periods (48-72 h), but higher concentrations (50-100 mM) can cause hypertonicity and reduced viability. In vivo, glycerophosphate administered intraperitoneally or intravenously in rodents at doses of 500-2000 mg/kg may cause transient hypocalcemia due to phosphate complexation with calcium, leading to neuromuscular irritability or tetany. However, such toxicities are due to phosphate overload, not specific to glycerophosphate. The alpha and beta mixture and the disodium salt form are not associated with genotoxicity, carcinogenicity, or reproductive toxicity. The compound is not intended for human or animal use as a drug. Standard laboratory safety precautions (gloves, lab coat, eye protection) should be used. The product is for research use only.

beta-Glycerophosphate (beta-GP) is a well-known component of osteogenic differentiation media. The alpha and beta isomers differ in the position of the phosphate group. The beta isomer has the phosphate on the middle carbon (C2) and is more commonly used in cell culture because it is more stable and less prone to non-enzymatic hydrolysis. The alpha isomer (phosphate on C1) is also known. The mixture is provided because both isomers may be present after synthesis. Glycerophosphoric acid is an endogenous metabolite: it is an intermediate in the metabolism of phospholipids (glycerophospholipids) and triglycerides. It is also a precursor for the synthesis of phospholipids in the Kennedy pathway. As a disodium salt, it provides sodium and phosphate. The hydrate contains a variable amount of water (x H2O), which is indicated in the formula. This product is a common laboratory chemical (CAS# 154804-51-0). It is not a drug and has no approved medical indication. It is for research use only.

C6H14NA4O12P2.XH2O

β-Glycerophosphate disodium salt hydrate;154804-51-0

Typically exists as solid at room temperature

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

H2O :~100 mg/mL

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.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*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).

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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)]
*Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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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).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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 (Please use freshly prepared in vivo formulations for optimal results.)