High-performance liquid chromatographic analysis of the new antitumour drug N-benzoylstaurosporine (CGP 41 251) and four potential metabolites in micro-volumes of plasma.
A high-performance liquid chromatographic (HPLC) assay is described for the determination of the new antitumour drug N-benzoylstaurosporine (CGP 41 251; I) and four of its potential metabolites in micro-volumes (100 microliters) of plasma. After addition of an internal standard, the compounds were isolated from plasma by liquid-liquid extraction with diisopropyl ether. Chromatography was carried out using a 5 microns LiChrospher C-18 end-capped column (125 x 4.0 mm i.d.) and binary gradient elution with acetonitrile and a triethylamine-containing phosphate buffer (pH 3.6) as solvents. Fluorimetric detection was performed with excitation and emission wavelengths set at 286 and 386 nm, respectively. The absolute recovery was more than 98% for all of the investigated compounds. The limit of detection (LOD) for I and three metabolites was 0.1 ng ml-1 and the lower limit of quantitation (LLQ) was 0.2 ng ml-1 in 100 microliters of plasma. The LOD and LLQ for the fourth metabolite was 0.25 and 0.5 ng ml-1, respectively. The between-day and within-day precisions were always < 15% for all the analytes. A limited pharmacokinetic study in mice treated and with I demonstrated that the method is appropriate for this purpose.
van Gijn
R
R
Department of Pharmacy, Slotervaart Hospital/Netherlands Cancer Institute, Amsterdam.
Havik
E
E
Boven
E
E
Vermorken
J B
JB
ten Bokkel Huinink
W W
WW
van Tellingen
O
O
Beijnen
J H
JH
D000818:Animals; D000970:Antineoplastic Agents / Q000097:blood* / Q000493:pharmacokinetics; D001711:Biotransformation; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D011786:Quality Control; D051381:Rats; D012015:Reference Standards; D012996:Solutions; D019311:Staurosporine / Q000031:analogs & derivatives* / Q000097:blood / Q000493:pharmacokinetics
D000970:Antineoplastic Agents; D012996:Solutions; D019311:Staurosporine; C059539:midostaurin