Schänzle G, Li S, Mikus G, Hofmann U
Dr. Margarete Fischer-Bosch-Institut für Klinische Pharmakologie, Stuttgart, Germany.
J Chromatogr B Biomed Sci Appl. 1999 Jan 8;721(1):55-65. doi: 10.1016/s0378-4347(98)00438-1.
A rapid, highly sensitive method for the determination of morphine and its metabolites morphine-3-glucuronide (M3G), morphine-6-glucuronide (M6G) and normorphine has been developed using high-performance liquid chromatography-electrospray mass spectrometry, with the deuterated analogues as internal standards. The analytes were extracted automatically using end-capped C2 solid-phase extraction cartridges. Baseline separation of morphine, M3G and M6G was achieved on a LiChrospher 100 RP-18 end-capped analytical column (125x3 mm I.D., 5 microm particle size) with water-acetonitrile-tetrahydrofuran-formic acid (100:1:1:0.1, v/v) as the mobile phase. Morphine and normorphine coeluate and were separated mass spectrometrically. The mass spectrometer was operated in the selected-ion monitoring mode using m/z 272 for normorphine, m/z 286 for morphine, m/z 462 for morphine-6-glucuronide. Due to an interfering peak, M3G was measured by tandem mass spectrometry in the daughter-ion mode. The limits of quantitation achieved with this method were 1.3 pmol/ml for morphine, 1.5 pmol/ml for normorphine, 1.0 pmol/ml for M6G and 5.4 pmol/ml for M3G in serum or cerebrospinal fluid. The limits of quantitation achieved in urine were 10 pmol/ml for morphine, 20 pmol/ml for normorphine and M6G and 50 pmol/ml for M3G using a sample size of 100 microl. The method described was successfully applied to the determination of morphine and its metabolites in human serum, cerebrospinal fluid and urine in pharmacokinetic and drug interaction studies.
已开发出一种快速、高灵敏度的方法，使用高效液相色谱 - 电喷雾质谱联用技术，并以氘代类似物作为内标，来测定吗啡及其代谢物吗啡 - 3 - 葡萄糖醛酸苷（M3G）、吗啡 - 6 - 葡萄糖醛酸苷（M6G）和去甲吗啡。使用封端的C2固相萃取柱自动萃取分析物。在LiChrospher 100 RP - 18封端分析柱（内径125x3 mm，粒径5微米）上，以水 - 乙腈 - 四氢呋喃 - 甲酸（100:1:1:0.1，v/v）作为流动相，实现了吗啡、M3G和M6G的基线分离。吗啡和去甲吗啡共洗脱，并通过质谱进行分离。质谱仪在选择离子监测模式下运行，使用m/z 272测定去甲吗啡，m/z 286测定吗啡，m/z 462测定吗啡 - 6 - 葡萄糖醛酸苷。由于存在干扰峰，M3G通过串联质谱在子离子模式下进行测定。该方法在血清或脑脊液中对吗啡的定量限为1.3 pmol/ml，对去甲吗啡为1.5 pmol/ml，对M6G为1.0 pmol/ml，对M3G为5.4 pmol/ml。在尿液中，使用100微升的样本量，吗啡的定量限为10 pmol/ml，去甲吗啡和M6G为20 pmol/ml，M3G为50 pmol/ml。所描述的方法已成功应用于药代动力学和药物相互作用研究中人体血清、脑脊液和尿液中吗啡及其代谢物的测定。
