Capillary electrophoresis stereoselective determination of R-(+)- and S-(-)-pentobarbital from serum using hydroxypropyl-gamma-cyclodextrin, solid-phase extraction and ultraviolet detection.

Pentobarbital is a sedative hypnotic which is used in the treatment of people suffering from anxiety related conditions and to manage elevated intracranial pressures and cerebral ischemia due to neurosurgical procedures. This paper develops and validates a rapid and sensitive method for the determination of S-(-)- and R-(+)-enantiomers in serum using capillary electrophoresis (CE) and UV detection. Stereoselective resolution was accomplished using 40 mM hydroxypropyl-gamma-cyclodextrin contained in 50 mM phosphate buffer pH 9.0. The method involves a solid-phase extraction of both the enantiomers and the internal standard, aprobarbital, from serum using C18 Bond-Elut cartridges. The CE system consists of a 52 cmX75 microm I.D. fused-silica capillary maintained at a run voltage of 15 kV with sample detection performed at 254 nm. The detection and quantitation limits for S-(-)- and R-(+)-pentobarbital are 1 microg/ml from serum. Linear calibration curves from 1 to 60 microg of both S-(-)- and R-(+)-enantiomers show a coefficient of determination of more than 0.999. The precision and accuracy of the method calculated as R.S.D. and error are 0.20-2.20% and 0.00-4.40%, respectively for R-(+)-pentobarbital and 0.30-2.19% and 0.30-6.40%, respectively for S-(-)-pentobarbital.