| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| Other Sizes |
| Targets |
5-Methyltetrahydrofolate is a cofactor for the enzyme methionine synthase (MS). It donates a methyl group to homocysteine to form methionine. This reaction is central to the methionine cycle and impacts many critical processes, including DNA methylation via S-adenosylmethionine (SAM). It also indirectly targets the enzymes involved in folate metabolism, such as methylenetetrahydrofolate reductase (MTHFR). It does not directly bind to a receptor but is a substrate for methionine synthase and a ligand for the reduced folate carrier (RFC).
|
|---|---|
| ln Vitro |
In vitro, 5-Methyltetrahydrofolic acid acts as a substrate, not an inhibitor. It is used in enzyme assays to measure the activity of methionine synthase. Its activity is measured by its ability to serve as a methyl donor in the conversion of homocysteine to methionine. In cell culture (e.g., fibroblasts), it is the predominant form of folate that enters the cell via the reduced folate carrier (RFC). It is a key regulator of intracellular folate pools and gene expression. It increases liver global DNA methylation relative to folic acid. No EC₅0 or IC₅0 values are applicable.
|
| ln Vivo |
In vivo, 5-Methyltetrahydrofolic acid is not a drug with direct activity but an essential nutrient that is absorbed, distributed, and metabolized. Dietary folates and folic acid are converted to 5-MTHF in the intestine and liver before being released into the circulation. It influences tissue-specific gene expression and increases liver global DNA methylation relative to folic acid. It is used for the management of folate deficiency and has been studied for its effect on cardiovascular risk via homocysteine lowering. It is the active form of folate used in some medical foods and supplements.
|
| Enzyme Assay |
Cell-free assays for 5-MTHF are enzyme assays. For methionine synthase, the enzyme is incubated with 5-Methyltetrahydrofolic acid, homocysteine, and a methyltransferase enzyme in a buffer (e.g., 50 mM potassium phosphate, pH 7.2). After incubation at 37degC, the reaction is stopped, and the product, methionine, is quantified by HPLC or by a radiolabeled method using [14C]-methyl-labeled 5-MTHF. The activity is measured by the rate of methionine formation, and the Michaelis-Menten constant (Km) for 5-MTHF can be determined. This is an enzyme kinetic study, not a binding assay.
|
| Cell Assay |
For cell-based studies, primary cells or cell lines (e.g., human fibroblasts) are cultured in folate-depleted medium. They are then incubated with varying concentrations of 5-Methyltetrahydrofolic acid (e.g., 0.1-100 nM). After 24-72 hours, cells are harvested. Intracellular folate polyglutamates are extracted and measured by HPLC or LC-MS/MS to assess uptake and metabolism. Global DNA methylation status can be assessed by using a methylated DNA quantification kit or by measuring the incorporation of [3H]-methyl from labeled 5-MTHF into DNA. These assays demonstrate its metabolic function.
|
| Animal Protocol |
In vivo studies are used to assess the bioavailability and physiological effects of 5-Methyltetrahydrofolic acid compared to folic acid (FA). In mouse models, animals are fed a diet with low folate. They are then supplemented with either folic acid or 5-MTHF at specific doses (e.g., 1-10 mg/kg diet) for several weeks. After the study period, blood is collected to measure serum folate and homocysteine levels. Tissues (liver, brain) are collected to measure folate content (LC-MS/MS) and DNA methylation levels (by HPLC or mass spectrometry). The endpoints are used to compare the efficacy of the two folate forms.
|
| ADME/Pharmacokinetics |
The pharmacokinetics of 5-Methyltetrahydrofolic acid are well-studied. It is absorbed in the proximal small intestine via the proton-coupled folate transporter (PCFT). It is the main transport form of folate in plasma. Once inside cells, it is converted to polyglutamate forms by folylpolyglutamate synthetase (FPGS) for tissue retention. The bioavailability of 5-MTHF is at least as high as that of folic acid. Its long-term intake increases both serum and red blood cell folate concentrations. An oral dose of 5-MTHF is absorbed with a Tmax of approximately 1-2 hours. Its half-life is not established.
|
| Toxicity/Toxicokinetics |
5-Methyltetrahydrofolic acid is generally recognized as safe (GRAS) and is the natural, active form of vitamin B9. It has a very low toxicity profile. Acute oral toxicity (LD₅0) is not established as it is a nutrient. High doses (e.g., 15 mg/day for several months) are generally well-tolerated. Potential side effects are mild and may include gastrointestinal upset. Unlike unmetabolized folic acid, 5-MTHF does not mask a vitamin B12 deficiency. The disodium salt form is water-soluble and is not considered a hazardous material under normal handling conditions. Standard laboratory safety practices are sufficient.
|
| References | |
| Additional Infomation |
5-Methyltetrahydrofolic acid disodium salt is not a drug and has no FDA/EMA approval for a specific therapeutic claim, though it is widely used as a dietary supplement and in medical foods. Its primary use is as a nutritional intervention to manage folate deficiency and hyperhomocysteinemia. It is used in research to study folate metabolism, one-carbon metabolism, and epigenetics. Unlike synthetic folic acid, 5-MTHF does not require metabolic activation and can be used in individuals with certain MTHFR polymorphisms. It is commercially available as a high-purity research standard for HPLC and LC-MS/MS analysis. It is stored at -20degC.
|
| Molecular Formula |
C20H23N7NA2O6
|
|---|---|
| Molecular Weight |
503.42
|
| Exact Mass |
503.151
|
| CAS # |
68792-52-9
|
| PubChem CID |
135445749
|
| Appearance |
Typically exists as solids at room temperature
|
| LogP |
0
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
10
|
| Rotatable Bond Count |
7
|
| Heavy Atom Count |
35
|
| Complexity |
854
|
| Defined Atom Stereocenter Count |
0
|
| SMILES |
[Na+].[Na+].O=C1C2=C(N=C(N([H])[H])N1[H])N([H])C([H])([H])C([H])(C([H])([H])N([H])C1C([H])=C([H])C(C(N([H])C([H])(C(=O)[O-])C([H])([H])C([H])([H])C(=O)[O-])=O)=C([H])C=1[H])N2C([H])([H])[H]
|
| InChi Key |
KKIWVYLOTHCGRV-UHFFFAOYSA-L
|
| InChi Code |
InChI=1S/C20H25N7O6.2Na/c1-27-12(9-23-16-15(27)18(31)26-20(21)25-16)8-22-11-4-2-10(3-5-11)17(30)24-13(19(32)33)6-7-14(28)29;;/h2-5,12-13,22H,6-9H2,1H3,(H,24,30)(H,28,29)(H,32,33)(H4,21,23,25,26,31);;/q;2*+1/p-2
|
| Chemical Name |
disodium;2-[[4-[(2-amino-5-methyl-4-oxo-3,6,7,8-tetrahydropteridin-6-yl)methylamino]benzoyl]amino]pentanedioate
|
| 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 | 1.9864 mL | 9.9321 mL | 19.8641 mL | |
| 5 mM | 0.3973 mL | 1.9864 mL | 3.9728 mL | |
| 10 mM | 0.1986 mL | 0.9932 mL | 1.9864 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
