{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,15]],"date-time":"2026-07-15T16:38:10Z","timestamp":1784133490137,"version":"3.55.0"},"reference-count":51,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2017,5,5]],"date-time":"2017-05-05T00:00:00Z","timestamp":1493942400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Nutrients"],"abstract":"<jats:p>Studies on one-carbon metabolism for the assessment of folate deficiency have focused on either metabolites of folate metabolism or methionine cycle. To bridge the gap between deficiency markers in these pathways we designed a dietary induced folate deficiency study using male C57BL\/6N mice. After weaning (3 weeks) mice were fed a defined control diet (1 week) before being fed a folate deficient diet (n = 6 mice) and the control diet (n = 6 mice) for 12 additional weeks. Thereafter, we determined total homocysteine in plasma and folate in erythrocytes as well as S-adenosylmethionine, S-adenosylhomocysteine, and six folate vitamers in tissues including 5-methyltetrahydrofolate, 5-formyltetrahydrofolate, 5,10-methenyltetrahydrofolate, tetrahydrofolate, 10-formylfolic acid, and folic acid by means of stable isotope dilution assays coupled with liquid chromatography tandem mass spectrometry. In all organs, except heart (mainly 5-mehtyltetrahydrofolate), tetrahydrofolate constitutes the main vitamer. Moreover, in liver tetrahydrofolate was most abundant followed by 5-methyltetrahydrofolate (heart: tetrahydrofolate), 5-formyltetrahydrofolate, and 5,10-methenyltetrahydrofolate. Because of the significant decrease (p &lt; 0.05) of folate status and S-adenosylmethionine\/S-adenosylhomocysteine ratio accompanied with increasing S-adenosylhomocysteine (p &lt; 0.05), hepatocytes are most susceptible to folate deficiency. To the best of our knowledge, we herein present the first method for simultaneous quantitation of eight metabolites for both folate and methionine cycle in one tissue sample, tHcy in plasma, and erythrocyte folate to shed light on physiological interrelations of one-carbon metabolism.<\/jats:p>","DOI":"10.3390\/nu9050462","type":"journal-article","created":{"date-parts":[[2017,5,5]],"date-time":"2017-05-05T10:31:08Z","timestamp":1493980268000},"page":"462","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":16,"title":["Characterization and Interrelations of One-Carbon Metabolites in Tissues, Erythrocytes, and Plasma in Mice with Dietary Induced Folate Deficiency"],"prefix":"10.3390","volume":"9","author":[{"given":"Markus","family":"Kopp","sequence":"first","affiliation":[{"name":"Chair of Analytical Food Chemistry, Technical University of Munich, Alte Akademie 10, Freising D-85354, Germany"},{"name":"Institute for Food &amp; Health (Z I E L), Technical University of Munich, Weihenstephaner Berg 1, Freising D-85354, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rosalie","family":"Morisset","sequence":"additional","affiliation":[{"name":"Institute for Food &amp; Health (Z I E L), Technical University of Munich, Weihenstephaner Berg 1, Freising D-85354, Germany"},{"name":"Chair of Nutritional Physiology, Technical University of Munich, Gregor-Mendel-Stra\u00dfe 2, Freising D-85354, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5826-6288","authenticated-orcid":false,"given":"Michael","family":"Rychlik","sequence":"additional","affiliation":[{"name":"Chair of Analytical Food Chemistry, Technical University of Munich, Alte Akademie 10, Freising D-85354, Germany"},{"name":"Centre for Nutrition and Food Sciences, Queensland Alliance for Agriculture and Food Innovation (QAAFI), University of Queensland, Brisbane, QLD 4072, Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2017,5,5]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1832","DOI":"10.1056\/NEJM199212243272602","article-title":"Prevention of the first occurrence of neural-tube defects by periconceptional vitamin supplementation","volume":"327","author":"Czeizel","year":"1992","journal-title":"N. Engl. J. Med."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1016\/j.reprotox.2015.07.073","article-title":"Preventable neural tube defects in Germany","volume":"56","author":"Obeid","year":"2015","journal-title":"Reprod. Toxicol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1136\/heart.83.2.127","article-title":"Homocysteine, B vitamins, and risk of cardiovascular disease","volume":"83","author":"Robinson","year":"2000","journal-title":"Heart"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"993","DOI":"10.1093\/ajcn\/71.4.993","article-title":"Serum folate and the severity of atrophy of the neocortex in Alzheimer disease: Findings from the Nun study","volume":"71","author":"Snowdon","year":"2000","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s00702-012-0822-y","article-title":"Folate and Alzheimer: When time matters","volume":"120","author":"Hinterberger","year":"2013","journal-title":"J. Neural Transm."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"55","DOI":"10.1111\/j.1365-2621.2004.tb17887.x","article-title":"The role of folate in reducing chronic and developmental disease risk: An overview","volume":"69","author":"Caudill","year":"2004","journal-title":"J. Food Sci."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"129","DOI":"10.1093\/jn\/130.2.129","article-title":"Folate and carcinogenesis: An integrated scheme","volume":"130","author":"Choi","year":"2000","journal-title":"J. Nutr."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"215","DOI":"10.3177\/jnsv.46.215","article-title":"Determination of folate derivatives in rat tissues during folate deficiency","volume":"46","author":"Nakata","year":"2000","journal-title":"J. Nutr. Sci. Vitaminol."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"986","DOI":"10.1093\/jn\/122.4.986","article-title":"Long-term folate deficiency alters folate content and distribution differentially in rat tissues","volume":"122","author":"Selhub","year":"1992","journal-title":"J. Nutr."},{"key":"ref_10","first-page":"67","article-title":"Effect of diet on folates levels and distribution in selected tissues of the rat","volume":"100","author":"Ladjimi","year":"1992","journal-title":"Arch. Int. Physiol. Biochim. Biophys."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1633","DOI":"10.1093\/jn\/120.12.1633","article-title":"Tissue distribution and prediction of total body folate of rats","volume":"120","author":"Clifford","year":"1990","journal-title":"J. Nutr."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1016\/S0304-3835(97)04708-3","article-title":"Alterations in hepatic p53 gene methylation patterns during tumor progression with folate\/methyl deficiency in the rat","volume":"115","author":"Pogribny","year":"1997","journal-title":"Cancer Lett."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"2811","DOI":"10.1093\/jn\/131.11.2811","article-title":"Intracellular S-adenosylhomocysteine concentrations predict global DNA hypomethylation in tissues of methyl-deficient cystathionine beta-synthase heterozygous mice","volume":"131","author":"Caudill","year":"2001","journal-title":"J. Nutr."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"2061","DOI":"10.1016\/j.jchromb.2009.05.039","article-title":"Determination of S-adenosylmethionine and S-adenosylhomocysteine by LC-MS\/MS and evaluation of their stability in mice tissues","volume":"877","author":"Krijt","year":"2009","journal-title":"J. Chromatogr. B Anal. Technol. Biomed. Life Sci."},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"Dahlhoff, C., Desmarchelier, C., Sailer, M., F\u00fcrst, R.W., Haag, A., Ulbrich, S.E., Hummel, B., Obeid, R., Geisel, J., and Bader, B.L. (2013). Hepatic methionine homeostasis is conserved in C57BL\/6N mice on high-fat diet despite major changes in hepatic one-carbon metabolism. PLoS ONE, 8.","DOI":"10.1371\/annotation\/9dd8c2df-1921-45b4-b3ba-8919f9068aea"},{"key":"ref_16","first-page":"4142","article-title":"Effect of 2\u2032-deoxycoformycin infusion on S-adenosylhomocysteine hydrolase and the amount of S-adenosylhomocysteine and related compounds in tissues of mice","volume":"43","author":"Helland","year":"1983","journal-title":"Cancer Res."},{"key":"ref_17","first-page":"1847","article-title":"S-adenosylhomocysteine and S-adenosylhomocysteine hydrolase in various tissues of mice given injections of 9-beta-d-arabinofuranosyladenine","volume":"43","author":"Helland","year":"1983","journal-title":"Cancer Res."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"521","DOI":"10.1042\/bj1660521","article-title":"Tissue distribution of S-adenosylmethionine and S-adenosylhomocysteine in the rat. Effect of age, sex and methionine administration on the metabolism of S-adenosylmethionine, S-adenosylhomocysteine and polyamines","volume":"166","author":"Eloranta","year":"1977","journal-title":"Biochem. J."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1371\/journal.pone.0156610","article-title":"Stable isotope dilution assays for clinical analyses of folates and other one-carbon metabolites: Application to folate-deficiency studies","volume":"11","author":"Kopp","year":"2016","journal-title":"PLoS ONE"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"36","DOI":"10.1152\/ajprenal.00385.2005","article-title":"The kidney in vitamin B12 and folate homeostasis: Characterization of receptors for tubular uptake of vitamins and carrier proteins","volume":"291","author":"Birn","year":"2006","journal-title":"Am. J. Physiol. Ren. Physiol."},{"key":"ref_21","unstructured":"Sachdeva, A., Yadav, S.P., and Arya, S.C. (2012). Nutritional Anemia. Practical Pediatric Hematology: Reference Manual for National Training Project, Jaypee Brothers Medical Publishers Ltd."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"3628","DOI":"10.1074\/jbc.272.6.3628","article-title":"Human methionine synthase. cDNA cloning, gene localization, and expression","volume":"272","author":"Chen","year":"1997","journal-title":"J. Biol. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"721","DOI":"10.1016\/0026-0495(75)90040-2","article-title":"Labile methyl balances for normal humans on various dietary regimens","volume":"24","author":"Mudd","year":"1975","journal-title":"Metab. Clin. Exp."},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Meister, A. (1979). Creatine: Biosynthesis, regulation and function. Advances in Enzymology and Related Areas of Molecular Biology, Wiley.","DOI":"10.1002\/9780470122945"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1151","DOI":"10.1093\/ajcn\/76.5.1151S","article-title":"S-adenosyl-l-methionine (SAMe): From the bench to the bedside\u2014Molecular basis of a pleiotrophic molecule","volume":"76","author":"Bottiglieri","year":"2002","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1694","DOI":"10.1515\/CCLM.2007.341","article-title":"Metabolic regulatory properties of S-adenosylmethionine and S-adenosylhomocysteine","volume":"45","author":"Finkelstein","year":"2007","journal-title":"Clin. Chem. Lab. Med."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"337","DOI":"10.3945\/ajcn.111.013367","article-title":"Folate status assessment history: Implications for measurement of biomarkers in NHANES","volume":"94","author":"Shane","year":"2011","journal-title":"Am. J. Clin. Nutr."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"1359","DOI":"10.1016\/0006-291X(82)91263-3","article-title":"The binding of folyl polyglutamates by hemoglobin","volume":"106","author":"Benesch","year":"1982","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"150","DOI":"10.1016\/j.ab.2009.11.007","article-title":"Quantitation of folates and their catabolites in blood plasma, erythrocytes, and urine by stable isotope dilution assays","volume":"398","author":"Netzel","year":"2010","journal-title":"Anal. Biochem."},{"key":"ref_30","unstructured":"Carmel, R., and Jacobsen, D.W. (2001). Historical Overview and Recent Perspective. Homocysteine in Health and Disease, Cambridge University Press."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1049","DOI":"10.1001\/jama.1995.03530130055028","article-title":"A quantitative assessment of plasma homocysteine as a risk factor for vascular disease. Probable benefits of increasing folic acid intakes","volume":"274","author":"Boushey","year":"1995","journal-title":"JAMA"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"31","DOI":"10.1146\/annurev.med.49.1.31","article-title":"Homocysteine and cardiovascular disease","volume":"49","author":"Refsum","year":"1998","journal-title":"Annu. Rev. Med."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"918","DOI":"10.1124\/jpet.108.143206","article-title":"Impact of high and low folate diets on tissue folate receptor levels and antitumor responses toward folate-drug conjugates","volume":"327","author":"Leamon","year":"2008","journal-title":"J. Pharm. Exp. Ther."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"469","DOI":"10.1111\/j.1365-2141.1975.tb01844.x","article-title":"Folate deficiency in the alcoholic\u2014Its relationship to clinical and haematological abnormalities, liver disease and folate stores","volume":"29","author":"Wu","year":"1975","journal-title":"Br. J. Haematol."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"13077","DOI":"10.1074\/jbc.271.22.13077","article-title":"Purification and properties of human cytosolic folylpoly-gamma-glutamate synthetase and organization, localization, and differential splicing of its gene","volume":"271","author":"Chen","year":"1996","journal-title":"J. Biol. Chem."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"530","DOI":"10.1021\/bi00376a027","article-title":"Mammalian folylpoly-gamma-glutamate synthetase. 4. In vitro and in vivo metabolism of folates and analogues and regulation of folate homeostasis","volume":"26","author":"Cook","year":"1987","journal-title":"Biochemistry"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"551","DOI":"10.1042\/cs0810551","article-title":"Betaine-homocysteine methyltransferase: Organ distribution in man, pig and rat and subcellular distribution in the rat","volume":"81","author":"McKeever","year":"1991","journal-title":"Clin. Sci."},{"key":"ref_38","first-page":"24","article-title":"Mechanisms of folate homeostasis","volume":"246","author":"Steinberg","year":"1984","journal-title":"Am. J. Physiol."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"85","DOI":"10.1007\/BF01622223","article-title":"Nahrungsfolate\u2014Eine aktuelle \u00dcbersicht Stabilit\u00e4t, Physiologische Bedeutung, Bioverf\u00fcgbarkeit, analytische Bestimmungsmethoden, Einflu\u00df der Lebensmittelbehandlung","volume":"33","author":"Pfeiffer","year":"1994","journal-title":"Zeitschrift Ern\u00e4hrungswissenschaft"},{"key":"ref_40","first-page":"228","article-title":"Methionine metabolism in mammals","volume":"1","author":"Finkelstein","year":"1990","journal-title":"J. Nutr. Physiol."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"4215","DOI":"10.1074\/jbc.M510624200","article-title":"Regulation of de novo purine biosynthesis by methenyltetrahydrofolate synthetase in neuroblastoma","volume":"281","author":"Field","year":"2006","journal-title":"J. Biol. Chem."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1002\/pca.1006","article-title":"pH stability of individual folates during critical sample preparation steps in prevision of the analysis of plant folates","volume":"18","author":"Zhang","year":"2007","journal-title":"Phytochem. Anal."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3067","DOI":"10.1021\/ja01151a028","article-title":"Synthetic and degradative investigations of the structure of folinic acid-SF","volume":"73","author":"May","year":"1951","journal-title":"J. Am. Chem. Soc."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"6500","DOI":"10.1073\/pnas.80.21.6500","article-title":"Evaluation of ascorbic acid in protecting labile folic acid derivatives","volume":"80","author":"Wilson","year":"1983","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1038\/ncomms3123","article-title":"Choroid plexus transcytosis and exosome shuttling deliver folate into brain parenchyma","volume":"4","author":"Grapp","year":"2013","journal-title":"Nat. Commun."},{"key":"ref_46","unstructured":"Carmel, R., and Jacobsen, D.W. (2001). Transport and tissue distribution of homocysteine and related S-adenosyl compounds. Homocysteine in Health and Disease, Cambridge University Press."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"231","DOI":"10.1016\/S0378-4347(98)00580-5","article-title":"Simultaneous determination of adenosine, S-adenosylhomocysteine and S-adenosylmethionine in biological samples using solid-phase extraction and high-performance liquid chromatography","volume":"724","author":"Delabar","year":"1999","journal-title":"J. Chromatogr. B Biomed. Sci. Appl."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"S40","DOI":"10.1007\/PL00014300","article-title":"The metabolism of homocysteine: Pathways and regulation","volume":"157","author":"Finkelstein","year":"1998","journal-title":"Eur. J. Pediatr."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"1707","DOI":"10.1096\/fj.06-7435com","article-title":"Mutations in methylenetetrahydrofolate reductase or cystathionine beta-synthase gene, or a high-methionine diet, increase homocysteine thiolactone levels in humans and mice","volume":"21","author":"Chwatko","year":"2007","journal-title":"FASEB J."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"29318","DOI":"10.1074\/jbc.M002725200","article-title":"Increase in plasma homocysteine associated with parallel increases in plasma S-adenosylhomocysteine and lymphocyte DNA hypomethylation","volume":"275","author":"Yi","year":"2000","journal-title":"J. Biol. Chem."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1501","DOI":"10.1002\/mnfr.201500991","article-title":"Gene specific epigenetic regulation of hepatic folate transport system is responsible for perturbed cellular folate status during aging and exogenous modulation","volume":"60","author":"Najar","year":"2016","journal-title":"Mol. Nutr. Food Res."}],"container-title":["Nutrients"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2072-6643\/9\/5\/462\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T18:34:48Z","timestamp":1760207688000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2072-6643\/9\/5\/462"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2017,5,5]]},"references-count":51,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2017,5]]}},"alternative-id":["nu9050462"],"URL":"https:\/\/doi.org\/10.3390\/nu9050462","relation":{},"ISSN":["2072-6643"],"issn-type":[{"value":"2072-6643","type":"electronic"}],"subject":[],"published":{"date-parts":[[2017,5,5]]}}}