Determination of three antiepileptic drugs in pharmaceutical formulations using microfluidic chips coupled with light-emitting diode induced fluorescence detection.

Abstract

In this study, a facile, sensitive, and precise lab-on-a-chip electrophoretic method coupled with light-emitting diode induced fluorescence (LED-IF) detection was developed to assay three antiepileptic drugs, namely, vigabatrin, pregabalin, and gabapentin, in pharmaceutical formulations. The analytes were derivatised offline for the first time with fluorescine-5-isothiocyanate (FITC) to yield highly fluorescent derivatives with λex/em of 490/520nm. The FITC-labelled analytes were injected, separated, and quantitated by a microfluidic electrophoresis device using fluorescence detection. The labelled analytes were monitored using a blue LED-IF system. The separation conditions were significantly optimised adding specific concentrations of heptakis-(2,6-di-O-methyl)-β-cyclodextrin (HDM-β-CD) and methylcellulose to the buffer solution (40mM borate buffer). HDM-β-CD acted as a selective host for the studied antiepileptic drugs, rendering a high separation efficiency. Methylcellulose was used as an efficient dynamic coating polymer to prevent the labelled drugs from being adsorbed on the inner surfaces of the poly (methylmethacrylate) microchannels. A laboratory-prepared ternary mixture of the three antiepileptic drugs was separated within 100s with acceptable resolution between all the three analytes (Rs>3) and a high number of theoretical plates (N) for each analyte (N≈106 plates/m). The sensitivity of the method was enhanced approximately 80-fold by stacking to yield a detection limit below 0.6ngmL-1 in the concentration range of 2.0-200.0ngmL-1. The method was successfully validated for analysing the studied drugs in their pharmaceutical formulations.