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Development and validation of a bioanalytical assay for (E)-5-(2-bromovinyl)-2'deoxyuridine in plasma by capillary
H J Reeuwijk1, U R Tjaden, J van der Greef
1Division of Analytical Chemistry, Leiden/Amsterdam Center for Drug Research, Leiden University, The Netherlands.
Abstract:
A capillary zone electrophoretic method for the quantification of (E)-5-(2-bromovinyl)-2'-deoxyuridine in plasma has been developed and validated. Separation was performed with a 25 mmol/l borate buffer, pH 9.0, after an initial rinsing step with sodium hydroxide. The rinsing step was necessary for reproducible analyses of aqueous samples and plasma extracts obtained by C18 solid-phase extraction after deproteination with perchloric acid. No interferences with plasma compounds were observed. The calibration graph was linear over the range of 30 to 3000 ng/ml using 5-fluorouracil as external standard. The limit of quantification was 24 ng/ml. The CZE method is fast, reproducible, linear and is therefore a good alternative for the already established HPLC methods.