Quenched phosphorescence as alternative detection mode in the chiral separation of methotrexate by electrokinetic chromatography.

M.G. Castro, I. Lammers, J.B. Buijs, C. Gooijer, F. Ariese

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Abstract

Quenched phosphorescence was used, for the first time, as detection mode in the chiral separation of methotrexate (MTX) enantiomers by electrokinetic chromatography. The detection is based on dynamic quenching of the strong emission of the phosphorophore 1-bromo-4-naphthalene sulfonic acid (BrNS) by MTX under deoxygenated conditions. The use of a background electrolyte with 3 mg/mL 2-hydroxypropyl-β-cyclodextrin and 20% MeOH in 25 mM phosphate buffer (pH 7.0) and an applied voltage of 30 kV allowed the separation of l-MTX and its enantiomeric impurity d-MTX with sufficient resolution. In the presence of 1 mM BrNS, a detection limit of 3.2 × 10
Original languageEnglish
Pages (from-to)2913-2919
JournalAnalytical and Bioanalytical Chemistry
Volume400
Issue number9
Early online date18 Apr 2011
DOIs
Publication statusPublished - 2011

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