A highly sensitive assay for the simultaneous determination of morphine, morphine-3-glucuronide, and morphine-6-glucuronide in human plasma by high-performance liquid chromatography with electrochemical and fluorescence detection.
A novel, highly sensitive and specific bioanalytical method has been developed for the simultaneous determination of morphine and its major metabolites, morphine-3-glucuronide and morphine-6-glucuronide, in human plasma, using noroxymorphone as the internal standard. The analytes are isolated from human plasma using a nonpolar/polar C2 solid-phase extraction cartridge and analyzed by high-performance liquid chromatography with serial detection using electrochemical detection for morphine, morphine-6-glucuronide (M6G), and noroxymorphone and fluorescence detection for morphine-3-glucuronide (M3G). The limit of quantitation (sensitivity) using a 0.5-ml sample of plasma is 1 ng/ml, 10 ng/ml, and 5 ng/ml for morphine, M3G, and M6G, respectively. Standard curves were linear (correlation coefficients > 0.999) over the ranges 1-30 ng/ml, 10-500 ng/ml, and 5-100 ng/ml for morphine, M3G, and M6G, respectively. The overall interday accuracy of the method was -1.58% for morphine, 2.27% for M3G, and -5.34% for M6G. The assay is routinely used for the study of morphine, M3G, and M6G pharmacokinetics after oral administration of morphine.
Rotshteyn
Y
Y
Department of Pharmacokinetics/Drug Metabolism, Purdue Frederick Company, Yonkers, New York, USA.
Weingarten
B
B
D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004355:Drug Stability; D009020:Morphine / Q000097:blood*; D009022:Morphine Derivatives / Q000097:blood*; D009294:Narcotics / Q000097:blood*
D009022:Morphine Derivatives; D009294:Narcotics; C035349:morphine-6-glucuronide; D009020:Morphine; C018108:morphine-3-glucuronide