Liquid chromatographic determination of ethambutol in serum samples based on intramolecular excimer-forming fluorescence derivatization.
A selective liquid chromatographic method has been developed for the assay of ethambutol in serum samples. The assay involves intramolecular excimer-forming derivatization with 4-(1-pyrene)butanoyl chloride (PBC) and isocratic reversed-phase chromatography with fluorescence detection. After acetonitrile-deproteinization of the serum sample, the derivatization reaction of ethambutol with PBC was completed within 30 min at 50 degrees C. N,N'-Diethylethylenediamine was used as an internal standard. The detection limit of ethambutol was 23 ng/ml serum, corresponding to 180 fmol on column at a signal-to-noise ratio of 3. The present method was selective enough to analyze ethambutol in rabbit serum without any tedious sample clean-up procedure because biogenic monoamines gave no peak in the chromatogram. The method was applicable to drug monitoring in rabbit serum.
Nakano
Yukitaka
Y
Department of Pharmacy, Saga University Hospital, 5-1-1 Nabeshima, Saga 849-8501, Japan.
Nohta
Hitoshi
H
Yoshida
Hideyuki
H
Todoroki
Kenichiro
K
Saita
Tetsuya
T
Fujito
Hiroshi
H
Mori
Masato
M
Yamaguchi
Masatoshi
M
D000818:Animals; D000995:Antitubercular Agents / Q000097:blood* / Q000493:pharmacokinetics; D004977:Ethambutol / Q000097:blood* / Q000493:pharmacokinetics; D005029:Ethylenediamines; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D011817:Rabbits; D012015:Reference Standards; D015203:Reproducibility of Results; D012996:Solutions; D013050:Spectrometry, Fluorescence
D000995:Antitubercular Agents; D005029:Ethylenediamines; D005456:Fluorescent Dyes; D007202:Indicators and Reagents; D012996:Solutions; C477544:N,N-diethylethylenediamine; D004977:Ethambutol