DETERMINATION OF MORPHINE, M-6-G, M-3-G, CODEINE, C-6-G AND 6-MAM IN BODY FLUIDS BY MEANS OF LC-APCI-MS|
Bogusz M.J., Maier R.D., Erkens M., Driessen S.
Institute of Forensic Medicine, Aachen University of Technology, D-52057 Aachen, Germany
A selective assay of morphine-3-glucuronide (M3G), morphine-6-glucuronide (M6G), morphine, codeine, codeine-6-glucuronide (C6G) and 6-monoacetylmorphine (6-MAM) is described. The 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 recovery ranged from 85 to 89% for each analyte. The separation was performed on ODS column in acetonitrile-ammonium formate buffer 50 mM, pH 3.0 (5:95), using flow rate gradient from 0.6 to 1.1 ml/min (total analysis time was 17 min). The quantitative analysis was done using deuterated analogues of each compound. The selected ion monitoring detection was applied: m/z 286 (for morphine, M3G-aglycone and M6G aglycone), 289 (for morphine-d3, M3G-d3-aglycone and M6G-d3-aglycone), 300 (for codeine and C6G-aglycone), 303 (for C6G-d3-aglycone), 306 (for codeine-d6), 328 (for 6-MAM), 334 (for 6-MAM-d6), 462 (for M3G and M6G), 465 (for M3G-d3 and M6G-d3), 476 (for C6G) and 479 (for C6G-d3). The limits of quantitation were: 1 µg/l for morphine, 2 µg/l for 6-MAM, 5 µg/l for M3G, M6G and codeine, 200 µg/l for C6G. The method appeared very selective and may be applied for routine determination of opiates in body fluids of heroin abusers and patients treated with opiates.