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The influence of 7-OH methotrexate metabolite on clinical relevance of methotrexate determination
E. Klapkova, J. Kukacka, K. Kotaska, I. Suchanska, R. Urinovska, R. Prusa,
Language English Country Germany
Document type Comparative Study, Journal Article, Validation Study
PubMed
21888024
Knihovny.cz E-resources
- MeSH
- Precursor Cell Lymphoblastic Leukemia-Lymphoma blood drug therapy MeSH
- Biotransformation MeSH
- Child MeSH
- Fluorescence Polarization Immunoassay MeSH
- Humans MeSH
- Methotrexate analogs & derivatives analysis blood pharmacokinetics MeSH
- Adolescent MeSH
- Young Adult MeSH
- Drug Monitoring methods MeSH
- Bone Neoplasms blood drug therapy MeSH
- Osteosarcoma blood drug therapy MeSH
- Antimetabolites, Antineoplastic blood pharmacokinetics MeSH
- Chromatography, High Pressure Liquid methods MeSH
- Check Tag
- Child MeSH
- Humans MeSH
- Adolescent MeSH
- Young Adult MeSH
- Publication type
- Journal Article MeSH
- Comparative Study MeSH
- Validation Study MeSH
BACKGROUND: A modified high performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of methotrexate (MTX) and its main metabolite 7-hydroxymethotrexate (7-OHMTX) and compared to the immunochemical fluorescence polarization immunoassay (FPIA2) determination of methotrexate. METHODS: Methotrexate was determined by HPLC with UV detection at 303 nm after precipitation of proteins with trichloroacetic acid. Fluorescence polarization immunoassays (FPIA2) of methotrexate were performed on the TDx FLx Immunoassay Analyzer. RESULTS: Our data indicate good correlation between methotrexate levels > 1 micromol/L determined by HPLC and FPIA2. (r = 0.94, Spearman correlation coefficient). However, concentrations of methotrexate < 1 micromol/L measured by fluorescence polarization immunoassay were overestimated. CONCLUSIONS: The concentration of MTX < 1 micromol/L are overestimated due to the cross reactivity with metabolites 7-OHMTX and 2,4-diamino-N10-methylpteroic acid (DAMPA). The cross reaction may affect the therapy and lead to relapse in children with acute lymphoblastic leukemia.
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- $a Klapkova, Eva $u Department of Clinical Biochemistry and Pathobiochemistry, Charles University, 2nd Faculty of Medicine and Faculty Hospital Motol, Prague, Czech Republic. eva.klapkova@email.cz
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- $a BACKGROUND: A modified high performance liquid chromatographic (HPLC) method has been developed for the simultaneous determination of methotrexate (MTX) and its main metabolite 7-hydroxymethotrexate (7-OHMTX) and compared to the immunochemical fluorescence polarization immunoassay (FPIA2) determination of methotrexate. METHODS: Methotrexate was determined by HPLC with UV detection at 303 nm after precipitation of proteins with trichloroacetic acid. Fluorescence polarization immunoassays (FPIA2) of methotrexate were performed on the TDx FLx Immunoassay Analyzer. RESULTS: Our data indicate good correlation between methotrexate levels > 1 micromol/L determined by HPLC and FPIA2. (r = 0.94, Spearman correlation coefficient). However, concentrations of methotrexate < 1 micromol/L measured by fluorescence polarization immunoassay were overestimated. CONCLUSIONS: The concentration of MTX < 1 micromol/L are overestimated due to the cross reactivity with metabolites 7-OHMTX and 2,4-diamino-N10-methylpteroic acid (DAMPA). The cross reaction may affect the therapy and lead to relapse in children with acute lymphoblastic leukemia.
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