Determination of 2-methyl derivatives of tamoxifen in cell culture medium using high-performance liquid chromatography and electrochemical detection.
The 2-methyl derivatives of tamoxifen (2-methyltamoxifen and 2-methyl-4-hydroxytamoxifen) were extracted from a cell culture medium at pH 5.4 (Earle's Minimum Essential Medium) with an internal standard (tamoxifen) on a phenyl sorbent cartridge. The compounds were then separated by high-performance liquid chromatography on a nitrile column eluted with acetonitrile-methanol-0.05 M sodium dihydrogenphosphate (19:4:11.6:69,v/v) containing 0.11 mmol/l disodium EDTA and determined by electrochemical detection at +1.1 V vs. Ag/AgCl/3 M NaCl. The absolute detection limits were 50 pg for 2-methyl-4-hydroxytamoxifen and 100 pg for tamoxifen and 2-methyltamoxifen at a sensitivity of 1 nA/V.
Chamart
S
S
Unité de Chimie Bioanalytique et de Toxicologie, Université Libre de Bruxelles, Belgium.
Hanocq
M
M
Helson
M
M
Devleeschauwer
N
N
Leclercq
G
G
D001943:Breast Neoplasms / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D003470:Culture Media; D004563:Electrochemistry; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009994:Osmolar Concentration; D013056:Spectrophotometry, Ultraviolet; D013629:Tamoxifen / Q000031:analogs & derivatives / Q000032:analysis* / Q000493:pharmacokinetics; D014407:Tumor Cells, Cultured / Q000378:metabolism
D003470:Culture Media; D013629:Tamoxifen