Coupling of on-Line Pre-Column Oxidative Cleavage and Solid-Phase Enrichment with Liquid Chromatography Using an Eco-Friendly Analytical Procedure to Determine Low Levels of Methotrexate.docVIP

Journal of Analytical Sciences, Methods and Instrumentation, 2012, 2, 194-202 /10.4236/jasmi.2012.24030 Published Online December 2012 (http://www.SciRP.org/journal/jasmi) Coupling of on-Line Pre-Column Oxidative Cleavage and Solid-Phase Enrichment with Liquid Chromatography Using an Eco-Friendly Analytical Procedure to Determine Low Levels of Methotrexate Samy Emara , Walaa Zarad , Maha Kamal , Ramzia EL-Bagary 1* 1 2 3 1 Pharmaceutical Chemistry Department, Faculty of Pharmacy, Misr International University, Cairo, Egypt; Pharmaceutical Analyti- 2 3 cal Chemistry Department, Faculty of Pharmacy, Modern Sciences and Arts University, 6th of October City Egypt; Pharmaceutical Chemistry Department, Faculty of Pharmacy, Cairo University, Giza, Egypt. Email: emara_miu@ * Received September 28 , 2012; revised November 9 , 2012; accepted November 18 , 2012 th th th ABSTRACT A simple, sensitive and precise green high-performance liquid chromatographic method including on-line pre-column oxidation combined by column switching with a short Hypersil ODS analytical column (100 mm × 4.0 mm i.d.) for enrichment and separation was developed and validated to determine low levels of methotrexate (MTX). The method was based on oxidative cleavage of MTX into highly fluorescence products, 2,4-diaminopteridine-6-carboxaldehyde and the corresponding 2,4-diaminopteridine-6-carboxylic acid, during the flow of phosphate buffer (0.04 M, pH 3.4) containing the analyte through the packed reactor of cerium (IV) trihydroxyhydroperoxide (CTH) at a flow-rate of 0.2 mL/min and 40?C. The fluorescent products were enriched on the head of ODS analytical column for the final separa- tio