Determination of WR-1065 in human blood by high-performance liquid chromatography following fluorescent derivatization by a maleimide reagent ThioGlo3.
In order to improve the sensitivity and stability of human blood samples containing WR-1065 (i.e., active metabolite of the cytoprotective agent amifostine), a high-performance liquid chromatographic method was developed and validated using fluorescent derivatization with ThioGlo3. Using a sample volume of only 100 microl, the method was specific, sensitive (limit of quantitation=10 nM in deproteinized blood or 20 nM in whole blood), accurate (error < or = 3.2%) and reproducible (CV < or = 8.7%). In addition, the stability of WR-1065 in deproteinized and derivatized blood samples was assured for at least four weeks at -20 degrees C. This method should be particularly valuable in translating the kinetic-dynamic relationship of WR-1065 in preclinical models to that in cancer patients.
Chen
Jun
J
Department of Pharmaceutical Sciences, College of Pharmacy, The University of Michigan, Ann Arbor, MI 48109, USA.
Lu
Zheng
Z
Lawrence
Theodore S
TS
Smith
David E
DE
D004999:Amifostine / Q000378:metabolism / Q000627:therapeutic use; D002851:Chromatography, High Pressure Liquid / Q000295:instrumentation / Q000379:methods*; D004355:Drug Stability; D006801:Humans; D008113:Liver Neoplasms / Q000188:drug therapy; D008624:Mercaptoethylamines / Q000097:blood*; D011714:Pyrans; D011758:Pyrroles; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013050:Spectrometry, Fluorescence
C434651:9-acetoxy-2-(4-(2,5-dihydro-2,5-dioxo-1H-pyrrol-1-yl)phenyl)-3-oxo-3H-naphtho(2,1-b)pyran; D008624:Mercaptoethylamines; D011714:Pyrans; D011758:Pyrroles; C020174:N-(2-mercaptoethyl)-1,3-diaminopropane; D004999:Amifostine