Stacking-cyclodextrin-microchip electrokinetic chromatographic determination of gabapentinoid drugs in pharmaceutical and biological matrices

- The first CD-MCEKC method was developed for the assay of gabapentinoids.
- The microfluidic channels were dynamically coated with methylcellulose.
- Cyclodextrins acted as pseudo-stationary phases to improve the method selectivity.
- Field-amplified stacking enhanced the sensitivity of the CD-MCEKC method.
- The Lab Chip CD-EKC was applied to pharmaceutical and biological analysis.

A facile, rapid, and highly sensitive microchip-based electrokinetic chromatographic method was developed for the simultaneous analysis of two gabapentinoid drugs, gabapentin (GPN) and pregabalin (PGN). Both drugs were first reacted with 4-fluoro-7-nitro-2,1,3-benzoxadiazole (NBD-F) via nucleophilic substitution reactions to yield highly fluorescent products with λex/em 470/540 nm. Analyses of both fluorescently labeled compounds were achieved within 200 s in a poly(methyl methacrylate) (PMMA) microchip with a 30 mm separation channel. Optimum separation was achieved using a borate buffer (pH 9.0) solution containing methylcellulose and β-cyclodextrin (β-CD) as buffer additives. Methylcellulose acted as a dynamic coating to prevent adsorption of the studied compounds on the inner surfaces of the microchannels, while β-CD acted as a pseudo-stationary phase to improve the separation efficiency between the labeled drugs with high resolution (Rs > 7). The fluorescence intensities of the labeled drugs were measured using a light emitting diode-induced fluorescence detector at 540 nm after excitation at 470 nm. The sensitivity of the method was enhanced 14- and 17-fold for PGN and GPN, respectively by field-amplified stacking relative to traditional pinched injection so that it could quantify 10 ng mL−1 for both analytes, with a detection limit lower than 3 ng mL−1. The developed method was efficiently applied to analyze PGN and GPN in their pharmaceutical dosage forms and in biological fluids. The extraction recoveries of the studied drugs from plasma and urine samples were more than 89% with%RSD values lower than 6.2.