Simultaneous measurement of the cell-differentiating agent hexamethylene bisacetamide and its metabolites by gas chromatography.
Hexamethylene bisacetamide (HMBA) is a potent in vitro differentiating agent that has clinical potential as an anticancer drug both as a single agent and as a component of combination therapy. A sensitive and efficient GC method for the isolation, derivatization, and measurement of both HMBA and its two major metabolites in plasma and urine in a single analysis is described. In situ carbamylation of the biological sample with diethylpyrocarbonate forms the urethane derivative of the basic N-acetyl diaminohexane metabolite and allows analyte isolation and concentration by solid-phase extraction. Subsequent formation of the n-butyl ester of 6-acetamidohexanoic acid, the major metabolite, provides a derivatized biological extract that can be rapidly analyzed by temperature-programmed GC. The quantitative extraction and the efficient derivatization steps provide a limit of quantitation of 0.05 mM (10 micrograms/ml) for all analytes with a precision better than 8% for the range of in vitro activity (0.1-2.0 mM). This method is amenable to automation and is well-suited for the analysis of clinical samples.
Roth
J S
JS
Laboratory of Medicinal Chemistry, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892-0037.
Kelley
J A
JA
Chun
H G
HG
Ward
F T
FT
D000081:Acetamides / Q000032:analysis* / Q000097:blood / Q000652:urine; D000614:Aminocaproates; D015119:Aminocaproic Acid / Q000097:blood / Q000652:urine; D000970:Antineoplastic Agents / Q000032:analysis* / Q000097:blood / Q000652:urine; D002849:Chromatography, Gas / Q000379:methods* / Q000706:statistics & numerical data; D004047:Diethyl Pyrocarbonate; D006801:Humans; D009369:Neoplasms / Q000097:blood / Q000188:drug therapy / Q000652:urine; D012680:Sensitivity and Specificity
D000081:Acetamides; D000614:Aminocaproates; D000970:Antineoplastic Agents; C050895:N-acetyl-1,6-diaminohexane; C018802:6-acetylaminocaproic acid; C014026:hexamethylene bisacetamide; D004047:Diethyl Pyrocarbonate; D015119:Aminocaproic Acid