Meesters Roland J W, den Boer Ethan, Mathot Ron A A, de Jonge Robert, van Klaveren Rob J, Lindemans Jan, Luider Theo M
Laboratories of Neuro-Oncology & Clinical & Cancer Proteomics, Department of Neurology, University Medical Center Rotterdam (ErasmusMC), Dr. Molewaterplein 50, Room Ee-1981, 3015 GE Rotterdam, The Netherlands.
Bioanalysis. 2011 Jun;3(12):1369-78. doi: 10.4155/bio.11.113.
A new analytical MS method using isotope dilution combined with MALDI-triple quadrupole MS/MS has been developed and validated for the determination of methotrexate and 7-hydroxymethotrexate in plasma. Methotrexate, methotrexate-d3, 7-hydroxymethotrexate and 7-hydroxymethotrexate-d3 were monitored by selected reaction monitoring using the transitions m/z 455.2→308.2, 458.2→311.2, 471.2→324.2 and 474.2→327.2 for methotrexate, methotrexate-d3, 7-hydroxymethotrexate and 7-hydroxymethotrexate-d3, respectively.
The LLOQ was 1 nmol/l for methotrexate and 7-hydroxymethotrexate while the limit of detection was 0.3 nmol/l for both analytes. The new developed method was cross-validated by a fluorescence polarization immunoassay and tested for its clinical feasibility by measuring plasma samples from patients suffering from acute lymphoblastic leukemia. Plasma methotrexate concentrations ranged between 66.0 and 954 nmol/l and observed 7-hydroxymethotrexate/methotrexate ratios ranged between 0.1 and 32.4, respectively.
The new method showed comparable analytical performances as the fluorescence polarization immunoassay, but analyte specificity and sensitivity of the newly developed method were significantly better.
