摘要
A new simple, sensitive and precise green analytical procedure using an automated packed-reactor derivatization technique coupled with on-line solid-phase enrichment （SPEn） has been developed and evaluated to determine trace levels of methotrexate （MTX）. The method was based on injection of MTX into a flowing stream of phosphate buffer （0.04 M, ptt 3.4）, carried through the packed oxidant reactor of Cerium （IV） trihydroxyhydroperoxide for oxidative cleavage of the drug into highly fluorescent product, 2,4-diaminopteridine- 6-carboxylic acid, followed by SPEn on a head of short ODS column （10mm x 4.6 mm i.d., 5 I＋tm particle size）. The flow rate was 0.25 mL/min and packed reactor temperaturc was 40 ~C. The trapped product was back-flush eluted from the ODS column to the detector by column-switching with an environmentally friendly mobile phase consisting of ethanol and phosphate buffer （0.04 M, pH 3.4） in the ratio of 5：95 （v/v）. The eluent was monitored at emission and excitation wavelengths of 460 and was linear over the concentration range of 1.25-50 360 nm, respectively. The calibration curve ng/mL with a detection limit of 0.08 ug/ml..The method was successfully applied to determine MTX in pharmaceutical formulations with mean percentage recovery ranging from 99.48 to 99.60.
A new simple, sensitive and precise green analytical procedure using an automated packed-reactor derivatization technique coupled with on-line solid-phase enrichment （SPEn） has been developed and evaluated to determine trace levels of methotrexate （MTX）. The method was based on injection of MTX into a flowing stream of phosphate buffer （0.04 M, ptt 3.4）, carried through the packed oxidant reactor of Cerium （IV） trihydroxyhydroperoxide for oxidative cleavage of the drug into highly fluorescent product, 2,4-diaminopteridine- 6-carboxylic acid, followed by SPEn on a head of short ODS column （10mm x 4.6 mm i.d., 5 I＋tm particle size）. The flow rate was 0.25 mL/min and packed reactor temperaturc was 40 ~C. The trapped product was back-flush eluted from the ODS column to the detector by column-switching with an environmentally friendly mobile phase consisting of ethanol and phosphate buffer （0.04 M, pH 3.4） in the ratio of 5：95 （v/v）. The eluent was monitored at emission and excitation wavelengths of 460 and was linear over the concentration range of 1.25-50 360 nm, respectively. The calibration curve ng/mL with a detection limit of 0.08 ug/ml..The method was successfully applied to determine MTX in pharmaceutical formulations with mean percentage recovery ranging from 99.48 to 99.60.
