Validated HPLC Method for Quantification of Pregabalin in Human Plasma Using 1-Fluoro-2,4-dinitrobenzene as Derivatization Agent

Abstract EN

In this study, a sensitive, simple, and reliable HPLC method for quantification of pregabalin in human plasma was developed and validated.

1-Fluoro-2,4-dinitrobenzene was used as precolumn derivatization agent.

For chromatography, an analytical reversed phase (C18) column and a mixture of Na2HPO4 10 mM (pH 8.0)—methanol (35 : 65 v/v) were used as stationary and mobile phase, respectively.

Detection was performed using a UV detector tuned at 360 nm.

The linearity of the method was tested over the concentration range 1–4500 ng/mL in 500 μL of human plasma and satisfactory results were obtained (r2 > 0.999).

The method showed good precision and accuracy in terms of within—between days relative standard deviations and percent deviation from nominated values (in the range of 4.3–12.7% and 2.6–8.0%, resp.).

The limit of quantification of the method was found to be 1 ng/mL which is better than previously reported method and indicates its potential application for sensitive bioanalysis.