Column-switching high-performance liquid chromatography-electrospray ionization mass spectrometry for identification of heroin metabolites in human urine
M. Katagi et al., Column-switching high-performance liquid chromatography-electrospray ionization mass spectrometry for identification of heroin metabolites in human urine, J CHROMAT B, 751(1), 2001, pp. 177-185
In order to prove heroin (DAM) use, a simple, rapid and sensitive analytica
l method has been established by combining semi-microcolumn HPLC, a column
switching technique and electrospray ionization mass spectrometry (ESI-MS).
Urine samples were directly introduced to the system, and endogenous urina
ry constituents were removed by using on-line column switching solid-phase
extraction with a strong cation-exchange (SCX) cartridge column (2.0 mm I.D
. X 10 mm). Heroin and its metabolites enriched on the top of the column we
re then successfully analyzed with excellent separation by use of a SCX sem
i-microcolumn (1.5 mm I.D. X 150 mm), accompanied by ESI mass spectral dete
ction. The proposed conditions are as follows: mobile phase, 10 mM ammonium
acetate (pH 6.0)-acetonitrile (30:70, v/v) (for main separation) and 30 mM
ammonium acetate (for trapping); flow-rates, 120 mul/min (for main separat
ion) and 200 mul/min (for trapping); capillary voltage, +4.5 kV; cone volta
ge, 50 V. Linear calibration curves were obtained in the selected ion monit
oring (SLM) mode using protonated molecular ions (m/z 370 for DAM, m/z 328
for MAM and mit 286 for MOR) over the concentration ranges from 10 to 1000
ng/ml for morphine (MOR) and 1-100 ng/ml for DAM and 6-acetylmorphine (MAM)
. The detection limits were 0.1-3 ng/ml. Upon applying the scan mode, 2-30
ng/ml were the detection limits. The present HPLC-ESI-MS method was success
fully applied to the determination of opiates in users' urine samples. (C)
2001 Elsevier Science B.V. All rights reserved.