{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,8,17]],"date-time":"2026-08-17T15:16:30Z","timestamp":1786979790320,"version":"build-2736575974"},"reference-count":35,"publisher":"Springer Science and Business Media LLC","issue":"20","license":[{"start":{"date-parts":[[2022,7,7]],"date-time":"2022-07-07T00:00:00Z","timestamp":1657152000000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,7,7]],"date-time":"2022-07-07T00:00:00Z","timestamp":1657152000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/501100001787","name":"University of South Australia","doi-asserted-by":"crossref","id":[{"id":"10.13039\/501100001787","id-type":"DOI","asserted-by":"crossref"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Anal Bioanal Chem"],"published-print":{"date-parts":[[2022,8]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>\n                    Low-dose methotrexate (MTX) plays a key role in treatment of rheumatoid arthritis. However, not all patients respond satisfactorily, and no therapeutic drug monitoring has been implemented in clinical practice, despite the fact that MTX therapy has now been available for decades. Analysis of individual intracellular MTX metabolites among rheumatoid arthritis (RA) patients is hampered by the low intracellular concentrations of MTX-PGs which require a highly sensitive method to quantify. Here, we present a rapid and highly sensitive LC (HILIC) MS\/MS method with LLOQ 0.1\u00a0nM, 0.8\u00a0nmol\/L for each metabolite of MTX-PG\n                    <jats:sub>1-5<\/jats:sub>\n                    and MTX-PG\n                    <jats:sub>6-7<\/jats:sub>\n                    respectively. Over a linear range of 0.1\u2013100\u00a0nM, 0.8\u2013100\u00a0nmol\/L for each metabolite of MTX-PG\n                    <jats:sub>1-5<\/jats:sub>\n                    and MTX-PG\n                    <jats:sub>6-7<\/jats:sub>\n                    , respectively, the inter- and intra- accuracy and precision were within 15% of the nominal value for all MTX metabolites. The presented assay was used to assess and compare MTX metabolite concentrations extracted from four different matrices: red blood cells, plasma, peripheral blood mononuclear cells, and whole blood that have been collected either using traditional venepuncture or volumetric absorptive micro-sampling (VAMS) sampling techniques. The presented method not only improves analyte coverage and sensitivity as compared to other published methods; it also improves the greenness.\n                  <\/jats:p>\n                  <jats:p>\n                    <jats:bold>Graphical abstract<\/jats:bold>\n                  <\/jats:p>","DOI":"10.1007\/s00216-022-04186-1","type":"journal-article","created":{"date-parts":[[2022,7,7]],"date-time":"2022-07-07T01:02:50Z","timestamp":1657155770000},"page":"6029-6046","update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Quantitation of methotrexate polyglutamates in human whole blood, erythrocytes and leukocytes collected via venepuncture and volumetric absorptive micro-sampling: a green LC\u2013MS\/MS-based method"],"prefix":"10.1007","volume":"414","author":[{"ORCID":"https:\/\/orcid.org\/0000-0003-1933-5241","authenticated-orcid":false,"given":"Dala N.","family":"Daraghmeh","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mahin","family":"Moghaddami","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Larisa","family":"Bobrovskaya","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Susanna M.","family":"Proudman","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michael D.","family":"Wiese","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2022,7,7]]},"reference":[{"issue":"5","key":"4186_CR1","doi-asserted-by":"publisher","first-page":"781","DOI":"10.1016\/S0738-081X(97)00014-X","volume":"15","author":"S Said","year":"1997","unstructured":"Said S, Jeffes EW, Weinstein GD. Methotrexate. Clin Dermatol. 1997;15(5):781\u201397.","journal-title":"Clin Dermatol"},{"issue":"3","key":"4186_CR2","doi-asserted-by":"publisher","first-page":"626","DOI":"10.1038\/leu.2017.283","volume":"32","author":"T Sakura","year":"2018","unstructured":"Sakura T, Hayakawa F, Sugiura I, Murayama T, Imai K, Usui N, et al. High-dose methotrexate therapy significantly improved survival of adult acute lymphoblastic leukemia: a phase III study by JALSG. Leukemia. 2018;32(3):626\u201332.","journal-title":"Leukemia"},{"issue":"4","key":"4186_CR3","doi-asserted-by":"publisher","first-page":"409","DOI":"10.1111\/j.1365-2125.1993.tb04158.x","volume":"35","author":"P Seideman","year":"1993","unstructured":"Seideman P, Beck O, Eksborg S, Wennberg M. The pharmacokinetics of methotrexate and its 7-hydroxy metabolite in patients with rheumatoid arthritis. Br J Clin Pharmacol. 1993;35(4):409\u201312.","journal-title":"Br J Clin Pharmacol"},{"key":"4186_CR4","doi-asserted-by":"crossref","unstructured":"Daraghmeh DN, King C, Wiese MD. A review of liquid biopsy as a tool to assess epigenetic, cfDNA and miRNA variability as methotrexate response predictors in patients with rheumatoid arthritis. Pharmacol Res. 2021;173:105887.","DOI":"10.1016\/j.phrs.2021.105887"},{"issue":"1","key":"4186_CR5","doi-asserted-by":"publisher","first-page":"28","DOI":"10.1016\/j.semarthrit.2015.02.009","volume":"45","author":"RB M\u00fcller","year":"2015","unstructured":"M\u00fcller RB, von Kempis J, Haile SR, Schiff MH. Effectiveness, tolerability, and safety of subcutaneous methotrexate in early rheumatoid arthritis: A retrospective analysis of real-world data from the St. Gallen cohort Semin Arthritis Rheum. 2015;45(1):28\u201334.","journal-title":"Gallen cohort. 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