RESUMEN
Chiral high performance liquid chromatography was used for the enantiometric separation of leucovorin. The optimal separation conditions recommended after optimizing the mobile phase composition, flow rate, and temperature are described. Achiral reversed-phase chromatographic method was applied for the simultaneous separation of methotrexate and leucovorin in clinical samples. Column-switching system including achiral short pre-column and chiral analytical column based on immobilized bovine serum albumin were used for simultaneous determination of leucovorin and methotrexate patients treated at the National Cancer Institute.
Asunto(s)
Leucovorina/química , Metotrexato/química , Cromatografía Líquida de Alta Presión , Humanos , Indicadores y Reactivos , Leucovorina/sangre , Leucovorina/aislamiento & purificación , Metotrexato/sangre , Metotrexato/aislamiento & purificación , EstereoisomerismoRESUMEN
Determination of polyamines (putrescine, spermidine and spermine) in urine samples of patients was worked out using high performance liquid chromatography (HPLC) and pre-column derivatization. For HPLC separation, a method of reversed-phase chromatography with gradient elution of water-organic mobile phases, UV and fluorescence detection were applied. Special care was devoted to the preseparation step, the hydrolysis, derivatization and selective extraction of clinical samples. The optimal HPLC conditions were recommended for simultaneous determination of putrescine, spermidine and spermine in a minimal analysis time (about 15 min) and suitable chromatographic resolution values (Rij greater than 1.5). Detection limits for all polyamines were evaluated (40 pmol for putrescine, 25 pmol for spermidine and 20 pmol for spermine) for the analysis of polyamines in the urine. The urine samples of healthy people and the cancer patients were compared and the procedure or the complete analysis has been described.