METHOD FOR MEASURING THE CONCENTRATION OF POLYGLUTAMATES OF METHOTREXATE BY HIGH-PERFORMANCE LIQUID CHROMATOGRAPHY WITH MASS SPECTROMETRIC DETECTION (HPLC-MS/MS)

摘要

FIELD: medicine.;SUBSTANCE: invention relates to a method for determining the concentration of methotrexate polyglutamates, including the collection of venous blood of a patient, separation of a sample of erythrocyte mass, isolation of methotrexate polyglutamates from it by means of a precipitation reaction and determination of the concentration of methotrexate polyglutamates using high-performance liquid chromatography with mass spectrometric detection (HPLC-MS/MS), moreover, the separated sample of the erythrocyte mass is frozen at a temperature of -80±2°C before the release of methotrexate polyglutamates from it, followed by thawing at a temperature of 6-10°C, while the precipitation reaction is carried out at a temperature of 6-10°C by introducing acidified acetonitrile into the sample, mixing it intensively, followed by centrifugation at 12000±10 rpm for 15±2 minutes and selecting the upper layer containing methotrexate polyglutamates for subsequent analysis using HPLC-MS/MS. ;EFFECT: wider range of methods for determining the concentration of methotrexate polyglutamates. ;8 cl, 1 dwg, 8 tbl, 4 ex