Simultaneous determination of heroin 6-monoacetylmorphine, morphine, and its glucuronides by liquid chromatography--atmospheric pressure ionspray-mass spectrometry.
A new analytical technique has been developed for the simultaneous determination of heroin, 6-monoacetylmorphine, morphine, morphine-6- and 3-glucuronides, and codeine in serum using liquid chromatography coupled with ionspray mass spectrometry. The analytes and the internal standard, nalorphine, were subjected to solid-phase extraction (SPE) using ethyl SPE columns before chromatography. The chromatographic separation of the analytes was achieved using a normal phase column and a water-methanol-acetonitrile-formic acid mobile phase at a flow rate of 230 microL/min. The mass spectrometer was operated in selected-ion monitoring mode. Under these conditions, the limit of quantitation was 0.5 ng/ml for heroin, 4 ng/ml for 6-monoacetylmorphine, 4 ng/ml for morphine, 1 ng/ml for morphine-3-glucuronide, 4 ng/ml for morphine-6-glucuronide, and 4 ng/mL for codeine. Serum levels of heroin metabolites were determined in C57BL/6 inbred mice after a dose of 20 mg/kg heroin administered subcutaneously. 6-monoacetylmorphine showed a peak concentration of 0.93 micrograms/mL serum at 3 min, whereas morphine and morphine-3-glucuronide achieved their peak concentrations of 9.6 and 2.9 micrograms/mL serum at 10 and 20 min, respectively. Finally, the absence of morphine-6-glucuronide and codeine excluded the possibility of their formation from morphine in this animal model.
Zuccaro
P
P
Clinical Biochemistry Department, Istituto Superiore di Sanità, Roma, Italy.
Ricciarello
R
R
Pichini
S
S
Pacifici
R
R
Altieri
I
I
Pellegrini
M
M
D'Ascenzo
G
G
D000818:Animals; D001274:Atmospheric Pressure; D003932:Heroin / Q000032:analysis / Q000378:metabolism / Q000493:pharmacokinetics; D013287:Illicit Drugs / Q000032:analysis*; D013058:Mass Spectrometry / Q000379:methods*; D051379:Mice; D008810:Mice, Inbred C57BL; D015394:Molecular Structure; D009020:Morphine / Q000032:analysis; D009022:Morphine Derivatives / Q000032:analysis; D009294:Narcotics / Q000032:analysis* / Q000378:metabolism / Q000493:pharmacokinetics; D015203:Reproducibility of Results
D013287:Illicit Drugs; D009022:Morphine Derivatives; D009294:Narcotics; C035349:morphine-6-glucuronide; D003932:Heroin; D009020:Morphine; C026979:6-O-monoacetylmorphine; C018108:morphine-3-glucuronide