Development and validation of HPLC method to determination of Methotrexate in children oncologic patients

OBJECTIVE: The acute lymphocytic leukemia is a hematopoietic cancer that occurs predominantly in children. Methotrexate is one of the most useful drugs in cancer chemotherapy. The aim of the study was to develop and validate the methodology of high performance liquid chromatography (HPLC) with ultraviolet detection for methotrexate dosage and to determine its concentration in plasma samples from children with leukemia.PATIENTS AND METHODS: The study included patients from the outpatient care of pediatric oncology at the Faculty of Medicine of ABC carriers in treatment of leukemia. The study was conducted in chromatographic model Agilent 1100 with UV detector at 302 nm and by the method of ELISA microplate reader capable of reading absorbance at 450 nm.RESULTS: We obtained satisfactory results of selectivity, accuracy, linearity, limit of quantification (LOQ), limit of detection (LOD), precision and robustness and apply the basic criteria for validation as RE No. 899, of May 29, 2003 Guide validation of analytical and bioanalytical National Agency Health Surveillance (ANVISA).CONCLUSIONS: We conclude that results for linearity/concentration range, precision, robustness, limit of quantification and detection limits are within the acceptance criteria defined by ANVISA and that the developed analytical method is valid and feasible to be used as a tool in monitoring therapy of methotrexate.Fac Med ABC, Lab Anal Clin, Santo Andre, SP, BrazilUniv Estadual Campinas, UNICAMP, Sch Med Sci FCM, Dept Pharmacol, Campinas, SP, BrazilUNIFESP, Inst Ciencias Ambientais Quim & Farmaceut, Diadema, SP, BrazilUNIFESP, Inst Ciencias Ambientais Quim & Farmaceut, Diadema, SP, BrazilWeb of Scienc