Identification and quantitation of methotrexate and methotrexate metabolites in clinical high-dose therapy by high-pressure liquid chromatography and field desorption mass spectrometry†

摘要: High-pressure liquid chromatography in combination with field desorption mass spectrometry as techniques of high specificity and sensitivity have been applied to the identification quantitation anticancer drug methotrexate its metabolites which occur clinical high-dose therapy. Field spectra several folic acid derivatives, when investigated free acids or ammonium salts, yield abundant protonated molecular ions a consistent pattern structurally significant fragments. chromatographic separation was performed on reverse-phase, C-18 columns using volatile, bicarbonate/acetate containing mobile phase that especially suited for spectral analysis isolated metabolites, provided definite 7-hydroxymethotrexate 4-[[2,4-diamino-6-pteridinyl]methyl]methylamino]-benzoic serum urine patients treated methotrexate. The intensity stability [MH]+ found suitable related folate analogues by spectrometry. A synthetic derivative, methotrexate-gamma-(2-hydroxy)ethyl-amide used internal standard quantitative determination urine. In study comparatively assess potential specific methods, levels major metabolite, were determined (i) an enzyme immunoassay, (ii) reverse high-pressure (iii) Results obtained from four osteogenic sarcoma receiving methotrexate/leucovorin rescue therapy consistently show sustained elimination over days, thus indicating utility specifically monitoring this nephrotoxic at massive doses.