Simple and sensitive spectrofluorimetric method for the determination of pregabalin in capsules through derivatization with fluorescamine

Abstract

A new, simple and sensitive spectrofluorimetric method has been developed for the determination of pregabalin (PG) in capsules. The method is based on the reaction between pregabalin and fluorescamine in borate buffer solution of pH 10 to give a highly fluorescent derivative that is measured at 487 nm after excitation at 390 nm. The different experimental parameters affecting the development and stability of the reaction product were carefully studied and optimized. The fluorescence intensity concentration plot was rectilinear over the range of 0.01-0.3 µg mL⁻¹ with a lower detection limit of 0.0017 µg mL⁻¹ and limit of quantitation of 0.005 µg mL⁻¹. The developed method was successfully applied to the analysis of the drug in its commercial capsules. The mean percentage recovery of PG in its capsule was 99.93±1.24 (n = 3). Statistical comparison of the results with those of the comparison method revealed good agreement and proved that there was no significant difference in the accuracy and precision of the two methods. A proposed reaction pathway was postulated.

Utility of Experimental Design in Pre-Column Derivatization for the Analysis of Tobramycin by HPLC—Fluorescence Detection: Application to Ophthalmic Solution and Human Plasma

“…Fluorescamine reagent is a useful derivatizing reagent that reacts with primary amino group to form fluorescent pyrrolinone moieties 27. Optimization of the pre-column derivatization step was performed using Design of Experiments (DOE) approach.…”

A Simple High-Throughput Method for Determination of Antiepileptic Analogues of γ-Aminobutyric Acid in Pharmaceutical Dosage Forms Using Microplate Fluorescence Reader

“…1) The analytical method development for PGB, GBP and VGB determination is challenging since these molecules have no significant UV or visible absorption. Several derivatization methods have been proposed [2][3][4][5][6][7][8][9][10][11][12][13][14][15][16][17][18][19] and used for their determination in bulk or pharmaceutical dosage forms, plasma, serum, and urine. These methods employed derivatization reagents such as fluorescamine, [2][3][4][17][18][19] 7-chloro-4-nitrobenzofurazan, [5][6][7] 1-fluoro-2,4-dinitrobenzene, 8) o-phtaldialdehyde, 9,10) 2,4,6-trinitrobenzene sulfonic acid, 11,12) and 9-fluorenylmethylchloroformate.…”

“…2,[5][6][7]17,19) Moreover, HPLC 3,[8][9][10][11][12][13][14]18) and capillary electrophoresis (CE) methods 4) using precolumn derivatization with either UV or fluorescent detector were also used.…”

“…In addition method should meet validation criteria in terms of precision, accuracy, robustness etc., and most of all it has to offer highthroughput. So far developed methods for determination of PGB, 3,5,17,19) GBP, 2,7,8,15,16,18,[20][21][22][23][24] and VGB 2,6,7,18,25) use from a few µL to approximately 1 mL of sample. The sample preparation is the time-consuming step in derivatization methods and in case of PGB, GBP, and VGB derivatization can take from few seconds to 50 min.…”

“…22,25) Cyclic voltammetry 23) and flow analysis with chemiluminometric detection 24) are rarely used for drug determination in pharmaceutical dosage forms and therefore less available for routine quality control analysis. All the above mentioned methods have been partially validated and proved to be suitable for quantification of PGB, GBP, and VGB in pharmaceutical dosage forms, however only a few methods were exposed to robustness test 3,17,[19][20][21][22] which is crucial for routine quality control analysis. Developed HPLC or CE methods have run times longer than 4 to 5 min, which does not allow a high-throughput analysis.…”