A high-performance liquid chromatography method using ultraviolet and fluorescence detection for the quantitation of UCN-01, 7-hydroxystaurosporine, from human plasma and saliva

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Abstract
UCN‐01, 7‐hydroxystaurosporine, is an antagonist of protein kinase C, as well as causing cell cycle arrest. We developed and validated an HPLC assay method for the quantitation of UCN‐01. Plasma and saliva standard curves were prepared at concentrations ranging from 0.2 to 20.0 µg/mL and 4.0 to 200.0 ng/mL, respectively. The sample preparation consisted of acetonitrile precipitation. Separation was accomplished on a phenyl column and a C‐18 precolumn insert utilizing a gradient‐profile consisting of ammonium acetate and acetonitrile. UV detection was set at 295 nm for UCN‐01 and 323 nm for umbelliferone, the internal standard. For fluorescence detection, excitation occurred at 290 nm, while emission was at 400 nm. The retention times were around 4 min for umbelliferone and 9.1 for UCN‐01. Inter‐ and intra‐assay errors of accuracy were less than 7.0% and 10.7%, respectively, for the plasma standard curve and less than 7.1% and 6.7%, respectively, for the saliva standard curve. The recoveries of UCN‐01 and umbelliferone from saliva were 81.4 ± 0.9% and 106.3 ± 10.2%, respectively. The recovery of UCN‐01 from plasma was 97.9 ± 7.1% and for umbelliferone was 103.3 ± 2.3%. This method is suitable for quantifying UCN‐01 in patient samples and further characterizing the clinical pharmacology of this compound. Published in 2000 by John Wiley & Sons, Ltd.