Determination of dexamethasone in plasma of premature neonates using high-performance liquid chromatography

A high-performance liquid chromatographic assay is described for dexamethasone in neonatal plasma. Samples (100 microliters) were extracted with ethyl acetate (1.0 ml) containing 50 ng of beclomethasone. The mobile phase of acetonitrile-0.01 M phosphate buffer (pH 7.0) (45:55, v/v) was pumped at a flow-rate of 1.0 ml/min through an Ultrasphere ODS column (5.0 microns particles). Detection was by UV absorbance at 240 nm. Calibration plots were linear (r2 > 0.999) from 10-1000 ng/ml. Within-day and between-day precision (C.V.%, coefficient of variation) between 15-900 ng/ml was 1.21-20.6%. Inaccuracy was 7.7%, or less. The minimum quantifiable concentration (C.V. approximately 20%) was 15 ng/ml. Recovery (20-1000 ng/ml) exceeded 75%. Concomitant neonatal medication did not interfere.