New spectrofluorimetric analysis of dapagliflozin after derivatization with NBD-Cl in human plasma using factorial design experiments.

Abstract

A new validated spectrofluorimetric method was proposed for dapagliflozin (DGF) analysis in bulk, plexin its commercially available tablets and in spiked human plasma. The proposed spectrofluorimetric method depended on the formation of a fluorescent complex soluble in organic liquids by a substitution reaction between 4-chloro-7-nitrobenzo-2-oxa-1,3-diazole (NBD-Cl) reagent and DGF in aqueous buffered solution at pH7. The fluorescence intensity was measured at 522nm after excitation at 453nm. The high selectivity of the proposed method allowed analysis of DGF in dosage form and human plasma samples with average recovery values of 99.84±1.38% and 98.71±1.80%, respectively, without any interference from matrix components. The calibration range was 50-1000ngml-1 . The limit of detection (LOD) and limit of quantitation (LOQ) were 14.24ngml-1 and 43.14ngml-1 , respectively. The estimated relative standard deviation values were lower than 2.0%, this showed the excellent precision at both levels. Factorial design was used to get the optimum method conditions for the analysis of the resulting DGF fluorescence complex in different matrices. The proposed method could be used in routine analysis of DGF in quality control laboratories. Also, it could be used to assay DGF in human plasma and be applied for pharmacokinetic investigation of DGF.