Journal
ELECTROPHORESIS
Volume 27, Issue 14, Pages 2905-2909Publisher
WILEY
DOI: 10.1002/elps.200500800
Keywords
CE; human plasma; large-volume sample stacking-EOF pump; metabolites; methotrexate
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This paper describes approaches for large-volume sample stacking (LVSS) with an EOF pump in CE for the determination of methotrexate (MTX) and its metabolites in human plasma. After pretreatment of plasma through a SPE cartridge, a large sample volume was loaded by hydrodynamic injection (3 psi, 70 s) into the capillary filled with phosphate buffer (70 mM, pH 6.0) containing 0.01% poly(ethylene oxide). Following removal of a large plug of sample matrix from the capillary using polarity switching (-25 kV), the separation of anionic analytes was subsequently performed without changing polarity again, achieving an improvement of sensitivity of around a 100-fold. The method was applied to therapeutic drug monitoring of MTX in one acute lymphoblastic leukemia patient. This study is one of very few applications showing the feasibility of LVSS in analysis of biological samples by CE.
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