SIMULTANEOUS ANALYSIS OF BENZPHETAMINE AND ITS METABOLITES, AND QUANTITATION OF URINARY P-HYDROXY-N-BENZYLAMPHETAMINE BY MICELLAR ELECTROKINETIC CHROMATOGRAPHY
A. Fujinami et al., SIMULTANEOUS ANALYSIS OF BENZPHETAMINE AND ITS METABOLITES, AND QUANTITATION OF URINARY P-HYDROXY-N-BENZYLAMPHETAMINE BY MICELLAR ELECTROKINETIC CHROMATOGRAPHY, Biological & pharmaceutical bulletin, 21(11), 1998, pp. 1207-1210
We developed a method for simultaneous analysis of benzphetamine (BZ)
and its metabolites, p-hydroxy-N-benzylamphetamine (pHBA), p-hydroxybe
nzphetamine (pHBz), amphetamine (AP), methamphetamine and p-hydroxymet
hamphetamine by micellar electrokinetic chromatography (MEKC). Urine s
amples from 0-15 h (3-h intervals) after oral administration of BZ (10
mg) were hydrolyzed with beta-glucuronidase (EC 3.2.1.31) at 37 degre
es C overnight. The treated urine was applied to a solid phase extract
ion column Bond Elut Certify(R). After sequentially washing the column
with water, 0.1 mol/l acetic acid and methanol, the samples were elut
ed with dichloromethane: isopropanol: 28% ammonium hydroxide=78.4 : 19
.6 : 2.0 (v/v %). The eluate was evaporated and the residue dissolved
in running buffer was analyzed by MEKC. In urine from 0-3 h, AP, pHBZ
and pHBA were detected. After that, only pHBA, which is one of the maj
or metabolites of BZ in human urine, could be detected in the urine by
the present method. A method for quantitation of pHBA by MEKC is desc
ribed here. The effects of acetonitrile and sodium dodecyl sulfate in
the running buffer of MEKC on the separation of BZ and its metabolites
are also reported.