Glibenclamide in serum: comparison of high-performance liquid chromatography using fluorescence detector and liquid chromatography/mass spectrometry with atmospheric-pressure chemical-ionization (APCI LC/MS)

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Abstract
Comparative studies of high-performance liquid Chromatographic and liquid chromatographic/mass spectrometric methods for the quantitative determination of glibenclamide in patient serum are described. The 4-methylcyclohexyl analogue of glibenclamide [N-(4-(β-5-chloro-2-methoxybenzamidoethyl)benzenesulfonyl-N′-(4-methyl-cyclohexyl)-urea] is used as internal standard for both methods. After acidification of the sample, the analyte and internal standard are extracted with chloroform. For HPLC analysis, the glibenclamide and internal standard are derivatized to a highly fluorescent amine with 7-chloro-4-nitrobenzo-2-oxa-l,3-diazole (NBD-chloride). The glibenclamide derivative is detected by a fluorescence detector. For APCI LC/MS assay, a derivatization of the analyte is not required and the compounds are measured by selected ion monitoring (SIM). The accuracy, precision, and recovery of the compared methods are presented and discussed.