Simple, rapid, and micro high-pressure liquid chromatographic determination of plasma griseofulvin levels

A rapid high-pressure liquid chromatographic assay is described for the quantitative determination of griseofulvin in plasma. An aliquot (25--10 microliter) of plasma was deproteinized by a simple procedure involving the addition of 2.5 volumes of acetonitrile, vortex mixing for a few seconds, and centrifugation for 1 min. The clear supernate, 50 microliter, was injected into the high-pressure liquid chromatograph. A reversed-phase column was used with a mobile phase of distilled water-acetonitrile (1:1) at a flow rate of 2 ml/min and was operated at ambient temperature. A fluorescent detector with an excitation wavelength of 260 nm was employed to monitor the column effluent. Griseofulvin had a retention time of 3.8 min. This procedure yields reproducible results with high sensitivity; plasma concentrations as low as 50 ng/ml can be measured. Several commonly used drugs do not interfere. Analysis of plasma samples collected from a rabbit injected with griseofulvin indicated that the procedure is suitable for pharmacokinetic studies and clinical monitoring of plasma concentrations in patients. Assay turnaround time is less than 6 min. For clinical monitoring of plasma griseofulvin concentrations, a sample volume as small as 10 microliter can be used.