The identification of in vitro metabolites of CKD-732 by liquid chromatography/tandem mass spectrometry.
The structural elucidation of fourteen metabolites of CKD-732, formed in vitro with rat liver microsomes, was performed using high-performance liquid chromatography/electrospray ionization-tandem mass spectrometry (HPLC/ESI-MS/MS). To identify proposed structures of the metabolites, the product ion mass spectra of the protonated molecules ([M + H]+), the retention times on reversed-phase HPLC, and UV-Vis spectra were utilized. Characteristic product ions for the identification of CKD-732 metabolites were observed at m/z 231, 236, and 252. The fragment ions at m/z 236 and 252 indicated the unchanged form and the N-oxide of the dimethylaminoethoxycinnamoyl group, respectively. The ion at m/z 231 indicated the presence of the hydroxylated form of the fumagillol group. The N-oxide of CKD-732, which was detected at m/z 515 and eluted later than CKD-732 in the reversed-phase HPLC system, was measured as a major metabolite. Three cis-trans isomers were also found.
Myung
Seung-Woon
SW
Doping Control Center, Korea Institute of Science and Technology, P.O. Box 131, Seoul 136-791, Korea. swmyung@kist.re.kr
Kim
Hye-Young
HY
Min
Hye-Ki
HK
Kim
Dong-Hyun
DH
Kim
Myungsoo
M
Cho
Hyun-Woo
HW
Lee
Ho Sup
HS
Kim
Joon-Kyum
JK
Hong
Chung I
CI
D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D003510:Cyclohexanes; D005231:Fatty Acids, Unsaturated / Q000032:analysis / Q000378:metabolism*; D008862:Microsomes, Liver / Q000378:metabolism; D051381:Rats; D012717:Sesquiterpenes; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods*
D000900:Anti-Bacterial Agents; D003510:Cyclohexanes; D005231:Fatty Acids, Unsaturated; D012717:Sesquiterpenes; C026211:fumagillin