DETERMINATION OF MORPHINE AND ITS 3-GLUCURONIDES AND 6-GLUCURONIDES, CODEINE, CODEINE-GLUCURONIDE AND 6-MONOACETYLMORPHINE IN BODY-FLUIDS BY LIQUID-CHROMATOGRAPHY ATMOSPHERIC-PRESSURE CHEMICAL-IONIZATION MASS-SPECTROMETRY

A selective assay of morphine-3-glucuronide (M3G), morphine-6-glucuron ide (M6G), morphine, codeine, codeine-6-glucuronide (C6G) and 6-monoac etylmorphine (6-MAM) based on liquid chromatography atmospheric pressu re chemical ionization mass spectrometry (LC-APCI-MS) is described. Th e drugs were extracted from serum, autopsy blood, urine, cerebrospinal fluid or vitreous humor using C-18 solid-phase extraction cartridges and subjected to LC-APCI-MS analysis. The separation was performed on an ODS column in acetonitrile-50 mM ammonium formate buffer, pH 3.0 (5 :95), using a flow-rate gradient from 0.6 to 1.1 ml/min (total analysi s time was 17 min). The quantitative analysis was done using deuterate d analogues of each compound. Selected-ion monitoring detection was ap plied: m/z 286 (for morphine, M3G-aglycone and M6G-aglycone), 289 (for morphine-d(3), M3G-d(3)-aglycone and M6G-d(3)-aglycone), 300 (for cod eine and C6G-aglycone), 303 (for C6G-d(3)-aglycone), 306 (for codeine- d(6)), 328 (for 6-MAM), 334 (for 6-MAM-d(6)), 462 (for M3G and M6G), 4 65 (for M3G-d(3) and M6G-d(3)), 476 (for C6G) and 379 (for C6G-d(3)). The limits of quantitation were: 1 mu g/l for morphine, 2 mu g/l for 6 -MAM, 5 mu g/l for M3G, M6G and codeine and 200 mu g/l for C6G. The re covery ranged from 85 to 98% for each analyte. The method appeared ver y selective and may be used for the routine determination of opiates i n body fluids of heroin abusers and patients treated with opiates. (C) 1997 Elsevier Science B.V.