Tomita Mamoru, Nakashima Mihoko N, Wada Mitsuhiro, Nakashima Kenichiro
Department of Clinical Pharmacy, Course of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
Biomed Chromatogr. 2007 Oct;21(10):1016-22. doi: 10.1002/bmc.839.
Simultaneous determination of 3,4-methylenedioxymethamphetamine (MDMA) and 3,4-methylenedioxyamphetamine (MDA) in rat blood and brain microdialysates by high-performance liquid chromatography with fluorescence detection (HPLC-FL) was developed. Microdialysates were directly subjected to derivatization with 4-(4,5-diphenyl-1H-imidazol-2-yl)benzoyl chloride (DIB-Cl). The DIB-derivatives of MDMA, MDA and the internal standard, 1-methyl-3-phenylpropylamine (MPPA), were isocratically separated on an ODS column using a mixture of 50 mm phosphate buffer (pH 7.0)-acetonitrile-methanol-2-propanol (50:45:5:2, v/v/v/v %) as an eluent at a flow rate of 1.5 mL/min. The calibration curves of MDA and MDMA spiked to blood and brain microdialysates were linear over the ranges 2.5-500 and 5.0-1000 ng/mL, respectively. The detection limits of MDA and MDMA were 1.2 and 4.2 for blood and 1.3 and 4.8 ng/mL for brain, respectively. Additionally, the intra- and the inter-assay precisions were lower than 5.6% for the blood and brain microdialysates (n = 4). The proposed method was successfully applied for the monitoring of MDMA and its metabolite MDA in rat blood and brain microdialysates, and the pharmacokinetic parameters of MDMA and MDA in the microdialysates after administration of MDMA (5 mg/kg, i.p.) with or without caffeine (20 mg/kg, i.p.) were evaluated.
建立了高效液相色谱荧光检测法（HPLC-FL）同时测定大鼠血液和脑微透析液中3,4-亚甲基二氧甲基苯丙胺（MDMA）和3,4-亚甲基二氧苯丙胺（MDA）的方法。微透析液直接用4-（4,5-二苯基-1H-咪唑-2-基）苯甲酰氯（DIB-Cl）进行衍生化。MDMA、MDA及其内标1-甲基-3-苯基丙胺（MPPA）的DIB衍生物在ODS柱上采用50 mM磷酸盐缓冲液（pH 7.0）-乙腈-甲醇-2-丙醇（50:45:5:2，v/v/v/v%）的混合物作为洗脱剂等度洗脱，流速为1.5 mL/min。加标到血液和脑微透析液中的MDA和MDMA的校准曲线在2.5-500和5.0-1000 ng/mL范围内分别呈线性。MDA和MDMA在血液中的检测限分别为1.2和4.2，在脑中的检测限分别为1.3和4.8 ng/mL。此外，血液和脑微透析液的批内和批间精密度均低于5.6%（n = 4）。该方法成功应用于大鼠血液和脑微透析液中MDMA及其代谢产物MDA的监测，并评估了MDMA（5 mg/kg，腹腔注射）单独或与咖啡因（20 mg/kg，腹腔注射）联合给药后微透析液中MDMA和MDA的药代动力学参数。
