Development of a high-performance liquid chromatographic method for the analysis of staurosporine.
Staurosporine (Stsp), a protein kinase inhibitor, has been found to have a differential effect on the proliferation of normal and transformed cells in vitro. Hence, Stsp might be used in cancer therapy to arrest normal proliferating cells in G1, while permitting tumor cells to continue proliferation. The patient could then be treated with a therapeutic agent of maximum toxicity for actively proliferating tumor cells. To facilitate investigations of Stsp in vivo, we have developed an HPLC method for measuring the levels of Stsp in blood. Using a rat model, plasma containing Stsp is treated with acetone to precipitate proteins and extract the Stsp. The acetone extract is then subjected to reversed-phase HPLC on a mu Bondapak C18 column. Using a linear elution gradient of acetonitrile containing trifluoroacetic acid, Stsp elutes as a sharp peak at ca. 35 min which can be detected by UV absorption at 292 nm. No blood or reagent components interfere with its quantification. The calibration curve, ranging from 0.1 to 2.0 micrograms Stsp, demonstrated a linear response to Stsp concentration having a correlation coefficient (r2) of 0.990. Precision analysis demonstrated that the method will yield results that are +/- 11.6% from the mean 95% (two standard deviations) of the time. This method was used to measure Stsp levels in plasma after administering an injection of 0.2 mg Stsp into the jugular vein of rats. No Stsp could be detected in the plasma 5 min after injection, even though enough Stsp was administered to be easily detectable if it was simply contained in the plasma. Thus, it is concluded that some compartment other than the plasma must adsorb the Stsp from the plasma and sequester it in vivo.
D000096:Acetone; D000470:Alkaloids / Q000097:blood*; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004492:Edetic Acid; D004791:Enzyme Inhibitors / Q000097:blood*; D006493:Heparin; D007202:Indicators and Reagents; D008297:Male; D011092:Polyethylene Glycols; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D051381:Rats; D011916:Rats, Inbred F344; D015203:Reproducibility of Results; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet; D019311:Staurosporine
D000470:Alkaloids; D004791:Enzyme Inhibitors; D007202:Indicators and Reagents; D012997:Solvents; D000096:Acetone; D011092:Polyethylene Glycols; D006493:Heparin; D004492:Edetic Acid; D011493:Protein Kinase C; D019311:Staurosporine