Determination of Benzodiazepines by Micellar Electrokinetic Chromatography

A method for the separation and determination of benzodiazepines by micellar electrokinetic chromatography (MEKC) has been developed. Separation buffers consisted of aqueous solutions of glycine and triethanolamine (pH 9.0), containing sodium dodecyl sulfate (SDS) as surfactant and methanol as organic modifier. The effect of the concentration of SDS, methanol, glycine and triethanolamine on migration times and resolution was studied. Ten benzodiazepines were baseline separated at a 25 mM SDS concentration and 20% v/v methanol in a 75 mM glycine-250 mM triethanolamine buffer. Under these conditions, the within-day reproducibilities were 0.3-0.5% for migration times and 1.7-1.9% for peak areas at a concentration of 10 micrograms/mL. The limits of detection and quantification for oxazepam were 0.2 and 0.7 micrograms/mL, respectively, using an injection time of 5 s.