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Record Information
StatusDetected and Quantified
Creation Date2005-11-16 15:48:42 UTC
Update Date2021-09-14 15:44:56 UTC
Secondary Accession Numbers
  • HMDB01396
Metabolite Identification
Common Name5-Methyltetrahydrofolic acid
Description5 methyltetrahydrofolic acid (5-MTHF) is the most biologically active form of the B-vitamin known as folic acid, also known generically as folate. 5-MTHF functions, in concert with vitamin B12, as a methyl-group donor involved in the conversion of the amino acid homocysteine to methionine. Methyl (CH3) group donation is vital to many bodily processes, including serotonin, melatonin, and DNA synthesis. Therapeutically, 5-MTHF is instrumental in reducing homocysteine levels, preventing neural tube defects, and improving vascular endothelial function. Research on folate supplementation suggests it plays a key role in preventing cervical dysplasia and protecting against neoplasia in ulcerative colitis. Folic acid also shows promise as part of a nutritional protocol to treat vitiligo, and may reduce inflammation of the gingiva. Furthermore, certain neurological, cognitive, and psychiatric presentations may be secondary to folate deficiency. Such presentations include depression, peripheral neuropathy, myelopathy, restless legs syndrome, insomnia, dementia, forgetfulness, irritability, endogenous depression, organic psychosis, and schizophrenia-like syndromes. After ingestion, the process of conversion of folic acid to the metabolically active coenzyme forms is relatively complex. Synthesis of the active forms of folic acid requires several enzymes, adequate liver and intestinal function, and adequate supplies of riboflavin (B2), niacin (B3), pyridoxine (B6), zinc, vitamin C, and serine. After formation of the coenzyme forms of the vitamin in the liver, these metabolically active compounds are secreted into the small intestine with bile (the folate enterohepatic cycle), where they are reabsorbed and distributed to tissues throughout the body. Human pharmacokinetic studies indicate folic acid has high bioavailability, with large oral doses of folic acid substantially raising plasma levels in healthy subjects in a time and dose dependent manner. Red blood cells (RBCs) appear to be the storage depot for folic acid, as RBC levels remain elevated for periods in excess of 40 days following discontinuation of supplementation. Folic acid is poorly transported to the brain and rapidly cleared from the central nervous system. The primary methods of elimination of absorbed folic acid are fecal (through bile) and urinary. Despite the biochemical complexity of this process, evidence suggests oral supplementation with folic acid increases the body's pool of 5-MTHF in healthy individuals. However, enzyme defects, mal-absorption, digestive system pathology, and liver disease can result in impaired ability to activate folic acid. In fact, some individuals have a severe congenital deficiency of the enzyme Methyl tetrahydrofolate reductase (5-MTHFR), which is needed to convert folic acid to 5-MTHF. Milder forms of this enzyme defect likely interact with dietary folate status to determine risk for some disease conditions. In individuals with a genetic defect of this enzyme (whether mild or severe), supplementation with 5- MTHF might be preferable to folic acid supplementation. (PMID: 17176169 ).
5-Methyl tetrahydrofolateHMDB
Methyl folateHMDB
N( 5)-MethyltetrahydrofolateHMDB
N-(4-(((2-Amino-1,4,5,6,7,8-hexahydro-5-methyl-4-oxo-6-pteridinyl)methyl)amino)benzoyl)-L-glutamic acidHMDB
N-(5-Methyl-5,6,7,8-tetrahydropteroyl)-L-glutamic acidHMDB
N5-Methyl-tetrahydrofolic acidHMDB
N5-Methyltetrahydropteroyl mono-L-glutamateHMDB
5-Methyltetrahydrofolate, (DL-glu)-isomerHMDB
5-Methyltetrahydrofolate, (L-glu)-(R)-isomerHMDB
5-Methyltetrahydrofolate, (L-glu)-(S)-isomerHMDB
5-Methyltetrahydrofolate, calcium salt (1:1), (L-glu)-isomerHMDB
5-Methyltetrahydrofolate, methyl-(14)C-labeled, (DL-glu)-isomerHMDB
5-Methyltetrahydrofolate, methyl-(14)C-labeled, (L-glu)-isomerHMDB
L-Methyl folateHMDB
N(5)-Methyltetrahydrofolic acidHMDB
Prefolic aHMDB
Levomefolic acidHMDB
Chemical FormulaC20H25N7O6
Average Molecular Weight459.4558
Monoisotopic Molecular Weight459.186631567
IUPAC Name(2R)-2-[(4-{[(2-amino-5-methyl-4-oxo-3,4,5,6,7,8-hexahydropteridin-6-yl)methyl]amino}phenyl)formamido]pentanedioic acid
Traditional Name(2R)-2-[(4-{[(2-amino-5-methyl-4-oxo-3,6,7,8-tetrahydropteridin-6-yl)methyl]amino}phenyl)formamido]pentanedioic acid
CAS Registry Number134-35-0
InChI Identifier
Chemical Taxonomy
Description Belongs to the class of organic compounds known as tetrahydrofolic acids. These are heterocyclic compounds based on the 5,6,7,8-tetrahydropteroic acid skeleton conjugated with at least one L-glutamic acid unit.
KingdomOrganic compounds
Super ClassOrganoheterocyclic compounds
ClassPteridines and derivatives
Sub ClassPterins and derivatives
Direct ParentTetrahydrofolic acids
Alternative Parents
  • Tetrahydrofolic acid
  • Glutamic acid or derivatives
  • Hippuric acid or derivatives
  • Hippuric acid
  • N-acyl-alpha-amino acid
  • N-acyl-alpha amino acid or derivatives
  • Alpha-amino acid or derivatives
  • Aminobenzamide
  • Aminobenzoic acid or derivatives
  • Benzoic acid or derivatives
  • Benzamide
  • Benzoyl
  • Phenylalkylamine
  • Tertiary aliphatic/aromatic amine
  • Aniline or substituted anilines
  • Dialkylarylamine
  • Secondary aliphatic/aromatic amine
  • Pyrimidone
  • Aminopyrimidine
  • Dicarboxylic acid or derivatives
  • Benzenoid
  • Pyrimidine
  • Monocyclic benzene moiety
  • Vinylogous amide
  • Heteroaromatic compound
  • Secondary carboxylic acid amide
  • Tertiary amine
  • Amino acid or derivatives
  • Amino acid
  • Carboxamide group
  • Secondary amine
  • Carboxylic acid
  • Carboxylic acid derivative
  • Azacycle
  • Organic nitrogen compound
  • Carbonyl group
  • Organonitrogen compound
  • Organooxygen compound
  • Primary amine
  • Hydrocarbon derivative
  • Organic oxide
  • Organopnictogen compound
  • Organic oxygen compound
  • Amine
  • Aromatic heteropolycyclic compound
Molecular FrameworkAromatic heteropolycyclic compounds
External DescriptorsNot Available
Physiological effect

Health effect:



Biological location:


Naturally occurring process:


Biological role:

Physical Properties
Experimental Molecular Properties
Melting PointNot AvailableNot Available
Boiling PointNot AvailableNot Available
Water SolubilityNot AvailableNot Available
LogPNot AvailableNot Available
Predicted Molecular Properties
Water Solubility0.35 g/LALOGPS
pKa (Strongest Acidic)3.22ChemAxon
pKa (Strongest Basic)4.64ChemAxon
Physiological Charge-2ChemAxon
Hydrogen Acceptor Count11ChemAxon
Hydrogen Donor Count7ChemAxon
Polar Surface Area198.48 ŲChemAxon
Rotatable Bond Count9ChemAxon
Refractivity126.88 m³·mol⁻¹ChemAxon
Polarizability46.13 ųChemAxon
Number of Rings3ChemAxon
Rule of FiveNoChemAxon
Ghose FilterNoChemAxon
Veber's RuleNoChemAxon
MDDR-like RuleYesChemAxon
Predicted Spectral Properties

Collision Cross Sections

NameAdductTypeData SourceValueReference
AllCCS[M-H]-ExperimentalNot Available197.7
AllCCS[M+H]+ExperimentalNot Available197.0
DarkChem[M+H]+PredictedNot Available206.33731661259
DarkChem[M-H]-PredictedNot Available198.05531661259
AllCCS[M+H]+PredictedNot Available203.77132859911
AllCCS[M-H]-PredictedNot Available199.27132859911

Retention Indices


Not Available


5-Methyltetrahydrofolic acid,1TMS,#14750.7144
5-Methyltetrahydrofolic acid,1TMS,#24745.3545
5-Methyltetrahydrofolic acid,1TMS,#34912.4893
5-Methyltetrahydrofolic acid,1TMS,#44733.9297
5-Methyltetrahydrofolic acid,1TMS,#54706.2676
5-Methyltetrahydrofolic acid,1TMS,#64628.061
5-Methyltetrahydrofolic acid,1TMS,#74797.4614
5-Methyltetrahydrofolic acid,2TMS,#14563.058
5-Methyltetrahydrofolic acid,2TMS,#24704.054
5-Methyltetrahydrofolic acid,2TMS,#34627.5786
5-Methyltetrahydrofolic acid,2TMS,#44427.149
5-Methyltetrahydrofolic acid,2TMS,#54518.4805
5-Methyltetrahydrofolic acid,2TMS,#64517.2275
5-Methyltetrahydrofolic acid,2TMS,#74711.6113
5-Methyltetrahydrofolic acid,2TMS,#84636.389
5-Methyltetrahydrofolic acid,2TMS,#94431.2515
5-Methyltetrahydrofolic acid,2TMS,#104515.643
5-Methyltetrahydrofolic acid,2TMS,#114543.52
5-Methyltetrahydrofolic acid,2TMS,#124756.8223
5-Methyltetrahydrofolic acid,2TMS,#134810.134
5-Methyltetrahydrofolic acid,2TMS,#144647.856
5-Methyltetrahydrofolic acid,2TMS,#154726.423
5-Methyltetrahydrofolic acid,2TMS,#164711.0537
5-Methyltetrahydrofolic acid,2TMS,#174639.7793
5-Methyltetrahydrofolic acid,2TMS,#184426.0674
5-Methyltetrahydrofolic acid,2TMS,#194478.22
5-Methyltetrahydrofolic acid,2TMS,#204605.244
5-Methyltetrahydrofolic acid,2TMS,#214393.4814
5-Methyltetrahydrofolic acid,2TMS,#224586.7344
5-Methyltetrahydrofolic acid,1TBDMS,#15004.5874
5-Methyltetrahydrofolic acid,1TBDMS,#24980.067
5-Methyltetrahydrofolic acid,1TBDMS,#35093.5117
5-Methyltetrahydrofolic acid,1TBDMS,#44958.335
5-Methyltetrahydrofolic acid,1TBDMS,#54934.2085
5-Methyltetrahydrofolic acid,1TBDMS,#64885.982
5-Methyltetrahydrofolic acid,1TBDMS,#75001.5483
5-Methyltetrahydrofolic acid,2TBDMS,#15035.6294
5-Methyltetrahydrofolic acid,2TBDMS,#25147.355
5-Methyltetrahydrofolic acid,2TBDMS,#35116.4155
5-Methyltetrahydrofolic acid,2TBDMS,#44912.701
5-Methyltetrahydrofolic acid,2TBDMS,#55008.3755
5-Methyltetrahydrofolic acid,2TBDMS,#64999.217
5-Methyltetrahydrofolic acid,2TBDMS,#75135.476
5-Methyltetrahydrofolic acid,2TBDMS,#85101.5566
5-Methyltetrahydrofolic acid,2TBDMS,#94913.51
5-Methyltetrahydrofolic acid,2TBDMS,#104994.656
5-Methyltetrahydrofolic acid,2TBDMS,#115001.6133
5-Methyltetrahydrofolic acid,2TBDMS,#125162.8447
5-Methyltetrahydrofolic acid,2TBDMS,#135216.2974
5-Methyltetrahydrofolic acid,2TBDMS,#145041.714
5-Methyltetrahydrofolic acid,2TBDMS,#155131.391
5-Methyltetrahydrofolic acid,2TBDMS,#165103.112
5-Methyltetrahydrofolic acid,2TBDMS,#175087.81
5-Methyltetrahydrofolic acid,2TBDMS,#184892.309
5-Methyltetrahydrofolic acid,2TBDMS,#194935.728
5-Methyltetrahydrofolic acid,2TBDMS,#205043.107
5-Methyltetrahydrofolic acid,2TBDMS,#214858.314
5-Methyltetrahydrofolic acid,2TBDMS,#225061.141


Spectrum TypeDescriptionSplash KeyDeposition DateView
Predicted GC-MSPredicted GC-MS Spectrum - GC-MS (Non-derivatized) - 70eV, Positivesplash10-03di-2645900000-0472550b7df0d95d47182017-09-01View Spectrum
Predicted GC-MSPredicted GC-MS Spectrum - GC-MS (2 TMS) - 70eV, Positivesplash10-059i-4532290000-8e3b1526c05a0a70d9712017-10-06View Spectrum
MSMass Spectrum (Electron Ionization)splash10-03di-2645900000-0472550b7df0d95d47182021-09-05View Spectrum


Spectrum TypeDescriptionSplash KeyDeposition DateView
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 10V, Positivesplash10-03dl-0313900000-d4f9d7f9d8c65a2e04282016-08-03View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 20V, Positivesplash10-01ox-0924300000-2f133db2f48bd5b14a8d2016-08-03View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 40V, Positivesplash10-000f-0920000000-298e8f3cd55b0fe42bad2016-08-03View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 10V, Negativesplash10-0a4i-0001900000-b1b1292a418283f61ea92016-08-03View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 20V, Negativesplash10-0900-1256900000-32c1cf48970dead32f602016-08-03View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 40V, Negativesplash10-0006-9641100000-b5468bf27455b7e5ee392016-08-03View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 10V, Negativesplash10-0a4i-0001900000-ba41f53e73f5102d5c042021-09-06View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 20V, Negativesplash10-0a4u-3419800000-385454b6189707f5e6842021-09-06View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 40V, Negativesplash10-0zg0-3931100000-ccab9c792ff1e540a4e82021-09-06View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 10V, Positivesplash10-03di-0108900000-f5ef3a5a71a48db8560b2021-09-07View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 20V, Positivesplash10-03di-0209100000-ac8dd1f905331a1f84bb2021-09-07View Spectrum
Predicted LC-MS/MSPredicted LC-MS/MS Spectrum - 40V, Positivesplash10-000x-0912000000-e973388138222d0175172021-09-07View Spectrum
Biological Properties
Cellular Locations
  • Extracellular
Biospecimen Locations
  • Blood
  • Cerebrospinal Fluid (CSF)
  • Urine
Tissue Locations
  • Kidney
  • Liver
  • Placenta
Normal Concentrations
BloodDetected but not QuantifiedNot QuantifiedAdult (>18 years old)BothNormal details
BloodDetected and Quantified0.3 (0.094-0.7) uMAdult (>18 years old)BothNormal details
Cerebrospinal Fluid (CSF)Detected and Quantified0.12 (0.04-0.20) uMAdult (>18 years old)BothNormal details
Cerebrospinal Fluid (CSF)Detected and Quantified0.0400-0.128 uMChildren (1-13 years old)BothNormal details
Cerebrospinal Fluid (CSF)Detected and Quantified0.075 (0.045 - 0.13) uMAdult (>18 years old)Not SpecifiedNormal details
UrineExpected but not QuantifiedNot QuantifiedNot AvailableNot AvailableNormal
      Not Available
UrineDetected but not QuantifiedNot QuantifiedAdult (>18 years old)BothNormal details
Abnormal Concentrations
Cerebrospinal Fluid (CSF)Detected and Quantified0.00700-0.0110 uMChildren (1-13 years old)BothCerebral folate transport deficiency details
Cerebrospinal Fluid (CSF)Detected and Quantified0.053 (0.02 - 0.09) uMAdult (>18 years old)Not SpecifiedRett syndrome details
Cerebrospinal Fluid (CSF)Detected and Quantified0.026 +/- 0.007 uMChildren (1-13 years old)Not Specified
Associated Disorders and Diseases
Disease References
Rett syndrome
  1. Ormazabal A, Artuch R, Vilaseca MA, Aracil A, Pineda M: Cerebrospinal fluid concentrations of folate, biogenic amines and pterins in Rett syndrome: treatment with folinic acid. Neuropediatrics. 2005 Dec;36(6):380-5. [PubMed:16429378 ]
  1. Surtees R, Hyland K, Smith I: Central-nervous-system methyl-group metabolism in children with neurological complications of HIV infection. Lancet. 1990 Mar 17;335(8690):619-21. [PubMed:1969014 ]
Cerebral folate transport deficiency
  1. Al-Baradie RS, Chaudhary MW: Diagnosis and management of cerebral folate deficiency. A form of folinic acid-responsive seizures. Neurosciences (Riyadh). 2014 Oct;19(4):312-6. [PubMed:25274592 ]
Associated OMIM IDs
DrugBank IDNot Available
Phenol Explorer Compound IDNot Available
FooDB IDFDB022600
KNApSAcK IDNot Available
Chemspider ID388371
KEGG Compound IDC00440
BioCyc IDNot Available
BiGG ID34988
Wikipedia LinkLevomefolic acid
PubChem Compound439234
PDB IDNot Available
ChEBI ID15641
Food Biomarker OntologyNot Available
MarkerDB IDMDB00000321
Synthesis ReferenceGennari, Federico. Process for producing 5-methyltetrahydrofolic acid and its salts. U.S. (1992), 6 pp.
Material Safety Data Sheet (MSDS)Not Available
General References
  1. Garbis SD, Melse-Boonstra A, West CE, van Breemen RB: Determination of folates in human plasma using hydrophilic interaction chromatography-tandem mass spectrometry. Anal Chem. 2001 Nov 15;73(22):5358-64. [PubMed:11816560 ]
  2. Ormazabal A, Artuch R, Vilaseca MA, Aracil A, Pineda M: Cerebrospinal fluid concentrations of folate, biogenic amines and pterins in Rett syndrome: treatment with folinic acid. Neuropediatrics. 2005 Dec;36(6):380-5. [PubMed:16429378 ]
  3. Chladek J, Sispera L, Martinkova J: High-performance liquid chromatographic assay for the determination of 5-methyltetrahydrofolate in human plasma. J Chromatogr B Biomed Sci Appl. 2000 Jul 21;744(2):307-13. [PubMed:10993519 ]
  4. Prasad PD, Mahesh VB, Leibach FH, Ganapathy V: Functional coupling between a bafilomycin A1-sensitive proton pump and a probenecid-sensitive folate transporter in human placental choriocarcinoma cells. Biochim Biophys Acta. 1994 Jun 30;1222(2):309-14. [PubMed:8031869 ]
  5. Kim TH, Yang J, Darling PB, O'Connor DL: A large pool of available folate exists in the large intestine of human infants and piglets. J Nutr. 2004 Jun;134(6):1389-94. [PubMed:15173401 ]
  6. Camilo E, Zimmerman J, Mason JB, Golner B, Russell R, Selhub J, Rosenberg IH: Folate synthesized by bacteria in the human upper small intestine is assimilated by the host. Gastroenterology. 1996 Apr;110(4):991-8. [PubMed:8613033 ]
  7. Kamen BA, Smith AK: A review of folate receptor alpha cycling and 5-methyltetrahydrofolate accumulation with an emphasis on cell models in vitro. Adv Drug Deliv Rev. 2004 Apr 29;56(8):1085-97. [PubMed:15094208 ]
  8. Surtees R, Leonard J, Austin S: Association of demyelination with deficiency of cerebrospinal-fluid S-adenosylmethionine in inborn errors of methyl-transfer pathway. Lancet. 1991 Dec 21-28;338(8782-8783):1550-4. [PubMed:1683972 ]
  9. Surtees R, Hyland K, Smith I: Central-nervous-system methyl-group metabolism in children with neurological complications of HIV infection. Lancet. 1990 Mar 17;335(8690):619-21. [PubMed:1969014 ]
  10. Evans MI, Duquette DA, Rinaldo P, Bawle E, Rosenblatt DS, Whitty J, Quintero RA, Johnson MP: Modulation of B12 dosage and response in fetal treatment of methylmalonic aciduria (MMA): titration of treatment dose to serum and urine MMA. Fetal Diagn Ther. 1997 Jan-Feb;12(1):21-3. [PubMed:9101216 ]
  11. Kane MA, Roth E, Raptis G, Schreiber C, Waxman S: Effect of intracellular folate concentration on the modulation of 5-fluorouracil cytotoxicity by the elevation of phosphoribosylpyrophosphate in cultured human KB cells. Cancer Res. 1987 Dec 15;47(24 Pt 1):6444-50. [PubMed:2445472 ]
  12. Irizarry MC: Biomarkers of Alzheimer disease in plasma. NeuroRx. 2004 Apr;1(2):226-34. [PubMed:15717023 ]
  13. Van Hove JL, Lazeyras F, Zeisel SH, Bottiglieri T, Hyland K, Charles HC, Gray L, Jaeken J, Kahler SG: One-methyl group metabolism in non-ketotic hyperglycinaemia: mildly elevated cerebrospinal fluid homocysteine levels. J Inherit Metab Dis. 1998 Dec;21(8):799-811. [PubMed:9870205 ]
  14. Straw JA, Szapary D, Wynn WT: Pharmacokinetics of the diastereoisomers of leucovorin after intravenous and oral administration to normal subjects. Cancer Res. 1984 Jul;44(7):3114-9. [PubMed:6609768 ]
  15. Authors unspecified: 5-methyltetrahydrofolate. Monograph. Altern Med Rev. 2006 Dec;11(4):330-7. [PubMed:17176169 ]


General function:
Involved in cobalamin binding
Specific function:
Catalyzes the transfer of a methyl group from methyl-cobalamin to homocysteine, yielding enzyme-bound cob(I)alamin and methionine. Subsequently, remethylates the cofactor using methyltetrahydrofolate (By similarity).
Gene Name:
Uniprot ID:
Molecular weight:
5-Methyltetrahydrofolic acid + Homocysteine → Tetrahydrofolic acid + L-Methioninedetails
General function:
Involved in methylenetetrahydrofolate reductase (NADPH) activity
Specific function:
Catalyzes the conversion of 5,10-methylenetetrahydrofolate to 5-methyltetrahydrofolate, a co-substrate for homocysteine remethylation to methionine.
Gene Name:
Uniprot ID:
Molecular weight:
5-Methyltetrahydrofolic acid + NAD(P)(+) → 5,10-Methylene-THF + NAD(P)Hdetails
5-Methyltetrahydrofolic acid + NADP → 5,10-Methylene-THF + NADPH + Hydrogen Iondetails
5-Methyltetrahydrofolic acid + NAD → 5,10-Methylene-THF + NADH + Hydrogen Iondetails
General function:
Not Available
Specific function:
N5-glutamine methyltransferase responsible for the methylation of the GGQ triplet of the mitochondrial translation release factor MTRF1L.
Gene Name:
Uniprot ID:
Molecular weight:
Not Available
Serotonin + 5-Methyltetrahydrofolic acid → 5-Methoxytryptamine + Tetrahydrofolic aciddetails
General function:
Not Available
Specific function:
Probable methyltransferase (By similarity).
Gene Name:
Uniprot ID:
Molecular weight:
Not Available
Serotonin + 5-Methyltetrahydrofolic acid → 5-Methoxytryptamine + Tetrahydrofolic aciddetails
General function:
Not Available
Specific function:
Probable methyltransferase (By similarity).
Gene Name:
Uniprot ID:
Molecular weight:
Not Available
Serotonin + 5-Methyltetrahydrofolic acid → 5-Methoxytryptamine + Tetrahydrofolic aciddetails
General function:
Not Available
Specific function:
Methyltransferase that may act on DNA.
Gene Name:
Uniprot ID:
Molecular weight:
Not Available
Serotonin + 5-Methyltetrahydrofolic acid → 5-Methoxytryptamine + Tetrahydrofolic aciddetails