Synonym
5-Methyltetrahydrofolate; 5-Methyltetrahydrofolic Acid; 5-methyl THF; Prefolic; Levomefolic acid; CH3-FH4; Methyl folate, N5-Methyl-tetrahydrofolic acid.
IUPAC/Chemical Name
N-[4-[[(2-amino-3,4,5,6,7,8-hexahydro-5-methyl-4-oxo-6-pteridinyl)methyl]amino]benzoyl]-L-glutamic acid
InChi Key
ZNOVTXRBGFNYRX-ABLWVSNPSA-N
InChi Code
InChI=1S/C20H25N7O6/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)/t12?,13-/m0/s1
SMILES Code
O=C(O)CC[C@@H](C(O)=O)NC(C1=CC=C(NCC2N(C)C3=C(N=C(N)NC3=O)NC2)C=C1)=O
Purity
>98% (or refer to the Certificate of Analysis)
Shipping Condition
Shipped under ambient temperature as non-hazardous chemical. This product is stable enough for a few weeks during ordinary shipping and time spent in Customs.
Storage Condition
Dry, dark and at 0 - 4 C for short term (days to weeks) or -20 C for long term (months to years).
Solubility
Soluble in DMSO
Shelf Life
>2 years if stored properly
Drug Formulation
This drug may be formulated in DMSO
Stock Solution Storage
0 - 4 C for short term (days to weeks), or -20 C for long term (months).
HS Tariff Code
2934.99.9001
Biological target:
5-Methyltetrahydrofolic acid (5-Methyl THF) is a biologically active form of folic acid and is a methylated derivate of tetrahydrofolate.
In vitro activity:
The aim of this study is to determine the effect of folates and vitamin B12 on endothelial vasoconstriction/vasodilatation parameters in cultured human endothelial cells incubated with human low density lipoproteins (LDL). Human umbilical vein endothelial cells (HUVEC) were extracted from recently delivered umbilical cords, cultured until confluence was achieved, and then incubated for 24h with folic acid (FA), 5-methyltetrahydrofolic acid (5-MTHF) or vitamin B12 (B12) in the presence or absence of LDL that was isolated from healthy volunteers. Total nitrites (as a measure of nitric oxide production), thiobarbituric acid reactive species (TBARS, a parameter of lipid peroxidation), and endothelin-1 (ET-1) were determined in the incubation media. None of the vitamins, either in the presence or absence of LDL, was able to modify nitric oxide production by HUVEC. A significant reduction of ET-1 production was observed in LDL-treated cells. This effect was not modified by FA or B12; however, 5-MTHF caused a concentration-dependent increase on ET-1 production, an effect coincidental with reduced TBARS production. This study demonstrates for the first time that 5MTHF, but not FA or B12, increases ET-1 production in LDL-treated endothelial cells. Although this effect was associated with the antioxidant properties of this folate, our results show that additional specific mechanisms involving 5-MTHF-LDL interactions may be operating to regulate endothelial function.
Reference: Nutr Metab Cardiovasc Dis. 2007 Mar;17(3):188-94. https://pubmed.ncbi.nlm.nih.gov/17367704/
In vivo activity:
Mice have been compeletly protected against the lethal effects of repeated injections of methotrexate by 5-methyltetrahydrofolic acid. Male C57BL/Bcr X IF/Bcr F1 hybrid mice were used throughout the experiment. In all experiments methotrexate was administered on 5 consecutive afternoons (days 1-5), and the test compounds were injected 5 hours earlier into the opposite flanks of the animals. While control mice dropped over 8 g. in weight before dying between 5 and 7 days after the first treatment with methotrexate, mice on the lower dose of 5-methyltetrahydrofolic acid dropped only 1*5 g. and then recovered. Subsequent experiments (ii, iii, v) showed that as little as 1 mg. of 5-methyltetrahydrofolic acid per kg. gave appreciable protection against methotrexate at 20 or 25 mg. per kg., while 3 mg. per kg. was completely protective.With a further doubling to 100 mg. of methotrexate per kg. (vi), however, some deaths still occurred even with 20 or 40 mg. of 5-methyltetrahydrofolic acid per kg. With three consecutive daily doses of 15 mg. per kg. (iii), the protective effect of 5-methyltetrahydrofolic acid was greatest (5/8 survivors) when given on days 3, 4 and 5 of methotrexate administration. Injections starting on days 1, 2, 4 or 5 failed to save more than 1 out of 8 animals. Reduction in the frequency of 5methyltetrahydrofolic acid administration thus greatly reduces its protective effect against methotrexate toxicity.
Reference: Br J Cancer. 1970 Sep;24(3):603-9. https://pubmed.ncbi.nlm.nih.gov/5312191/
|
Solvent |
mg/mL |
mM |
comments |
| Solubility |
| DMSO |
80.0 |
174.10 |
|
Note: There can be variations in solubility for the same chemical from different vendors or different batches from the same vendor. The following factors can affect the solubility of the same chemical: solvent used for crystallization, residual solvent content, polymorphism, salt versus free form, degree of hydration, solvent temperature. Please use the solubility data as a reference only. Warming and sonication will facilitate dissolving. Still have questions? Please contact our Technical Support scientists.
Preparing Stock Solutions
The following data is based on the
product
molecular weight
459.46
Batch specific molecular weights may vary
from batch to batch
due to the degree of hydration, which will
affect the solvent
volumes required to prepare stock solutions.
| Concentration / Solvent Volume / Mass |
1 mg |
5 mg |
10 mg |
| 1 mM |
1.15 mL |
5.76 mL |
11.51 mL |
| 5 mM |
0.23 mL |
1.15 mL |
2.3 mL |
| 10 mM |
0.12 mL |
0.58 mL |
1.15 mL |
| 50 mM |
0.02 mL |
0.12 mL |
0.23 mL |
Formulation protocol:
1. Ronco AM, Llanos M, Tamayo D, Hirsch S. 5-methyltetrahydrofolic acid stimulates endothelin-1 production in low density lipoprotein-treated human endothelial cells. Nutr Metab Cardiovasc Dis. 2007 Mar;17(3):188-94. doi: 10.1016/j.numecd.2005.12.005. Epub 2006 Mar 20. PMID: 17367704. 2. Fujii K, Nagasaki T, Huennekens FM. Accumulation of 5-methyltetrahydrofolate in cobalamin-deficient L1210 mouse leukemia cells. J Biol Chem. 1982 Mar 10;257(5):2144-6. PMID: 7061412.
3. Blair JA, Searle CE. Reversal of methotrexate toxicity in mice by 5-methyltetrahydrofolic acid. Br J Cancer. 1970 Sep;24(3):603-9. doi: 10.1038/bjc.1970.73. PMID: 5312191; PMCID: PMC2008620.
In vitro protocol:
1. Ronco AM, Llanos M, Tamayo D, Hirsch S. 5-methyltetrahydrofolic acid stimulates endothelin-1 production in low density lipoprotein-treated human endothelial cells. Nutr Metab Cardiovasc Dis. 2007 Mar;17(3):188-94. doi: 10.1016/j.numecd.2005.12.005. Epub 2006 Mar 20. PMID: 17367704. 2. Fujii K, Nagasaki T, Huennekens FM. Accumulation of 5-methyltetrahydrofolate in cobalamin-deficient L1210 mouse leukemia cells. J Biol Chem. 1982 Mar 10;257(5):2144-6. PMID: 7061412.
In vivo protocol:
1. Blair JA, Searle CE. Reversal of methotrexate toxicity in mice by 5-methyltetrahydrofolic acid. Br J Cancer. 1970 Sep;24(3):603-9. doi: 10.1038/bjc.1970.73. PMID: 5312191; PMCID: PMC2008620.
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2: Obeid R, Holzgreve W, Pietrzik K. Is 5-methyltetrahydrofolate an alternative to folic acid for the prevention of neural tube defects? J Perinat Med. 2013 Sep 1;41(5):469-83. doi: 10.1515/jpm-2012-0256. Review. PubMed PMID: 23482308.
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5: 5-methyltetrahydrofolate. Monograph. Altern Med Rev. 2006 Dec;11(4):330-7. Review. PubMed PMID: 17176169.
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7: Molero-Luis M, Serrano M, O'Callaghan MM, Sierra C, Pérez-Dueñas B, García-Cazorla A, Artuch R. Clinical, etiological and therapeutic aspects of cerebral folate deficiency. Expert Rev Neurother. 2015;15(7):793-802. doi: 10.1586/14737175.2015.1055322. Epub 2015 Jun 19. Review. PubMed PMID: 26092490.
8: Thase ME. Managing medical comorbidities in patients with depression to improve prognosis. J Clin Psychiatry. 2016 Feb;77 Suppl 1:22-7. doi: 10.4088/JCP.14077su1c.04. Review. PubMed PMID: 26829434.
9: Miller AL. The methylation, neurotransmitter, and antioxidant connections between folate and depression. Altern Med Rev. 2008 Sep;13(3):216-26. Review. PubMed PMID: 18950248.
10: Nzila A, Okombo J, Molloy AM. Impact of folate supplementation on the efficacy of sulfadoxine/pyrimethamine in preventing malaria in pregnancy: the potential of 5-methyl-tetrahydrofolate. J Antimicrob Chemother. 2014 Feb;69(2):323-30. doi: 10.1093/jac/dkt394. Epub 2013 Oct 14. Review. PubMed PMID: 24126794.
11: Gordon N. Cerebral folate deficiency. Dev Med Child Neurol. 2009 Mar;51(3):180-2. doi: 10.1111/j.1469-8749.2008.03185.x. Review. PubMed PMID: 19260931.
12: Seremak-Mrozikiewicz A. [Metafolin--alternative for folate deficiency supplementation in pregnant women]. Ginekol Pol. 2013 Jul;84(7):641-6. Review. Polish. PubMed PMID: 24032278.
13: Trimmer EE. Methylenetetrahydrofolate reductase: biochemical characterization and medical significance. Curr Pharm Des. 2013;19(14):2574-93. Review. PubMed PMID: 23116396.
14: Fava M, Mischoulon D. Folate in depression: efficacy, safety, differences in formulations, and clinical issues. J Clin Psychiatry. 2009;70 Suppl 5:12-7. doi: 10.4088/JCP.8157su1c.03. Review. PubMed PMID: 19909688.
15: Papakostas GI, Cassiello CF, Iovieno N. Folates and S-adenosylmethionine for major depressive disorder. Can J Psychiatry. 2012 Jul;57(7):406-13. Review. PubMed PMID: 22762295.
16: Strandler HS, Patring J, Jägerstad M, Jastrebova J. Challenges in the determination of unsubstituted food folates: impact of stabilities and conversions on analytical results. J Agric Food Chem. 2015 Mar 11;63(9):2367-77. doi: 10.1021/jf504987n. Epub 2015 Feb 26. Review. PubMed PMID: 25642846.
17: Bhatia P, Singh N. Homocysteine excess: delineating the possible mechanism of neurotoxicity and depression. Fundam Clin Pharmacol. 2015 Dec;29(6):522-8. doi: 10.1111/fcp.12145. Epub 2015 Sep 17. Review. PubMed PMID: 26376956.
18: Födinger M, Hörl WH, Sunder-Plassmann G. Molecular biology of 5,10-methylenetetrahydrofolate reductase. J Nephrol. 2000 Jan-Feb;13(1):20-33. Review. PubMed PMID: 10720211.
19: Hyland K, Shoffner J, Heales SJ. Cerebral folate deficiency. J Inherit Metab Dis. 2010 Oct;33(5):563-70. doi: 10.1007/s10545-010-9159-6. Epub 2010 Jul 29. Review. PubMed PMID: 20668945.
20: Saini RK, Nile SH, Keum YS. Folates: Chemistry, analysis, occurrence, biofortification and bioavailability. Food Res Int. 2016 Nov;89(Pt 1):1-13. doi: 10.1016/j.foodres.2016.07.013. Epub 2016 Jul 25. Review. PubMed PMID: 28460896.
