title: Determination of MDMA and MDA in rat urine by semi-micro column HPLC-fluorescence detection with DBD-F and their monitoring after MDMA administration to rat.
abstract: A simultaneous semi-micro column HPLC method with fluorescence detection of abused drugs, such as 3,4-methylenedioxymethamphetamine (MDMA), 3,4-methylenedioxyamphetamine (MDA), amphetamine (AP) and methamphetamine (MP) in rat urine was examined by using 4-(N,N-dimethylaminosulphonyl)-7-fluoro-1,2,3-benzoxadiazole (DBD-F) as a labelling reagent and alpha-phenylethylamine as an internal standard (IS). A sample (50 microL) of rat urine was added to 5 microL IS and 100 microL 100 mmol/L borate buffer (pH 12) and extracted with 1.5 mL n-hexane. After evaporation, 50 microL 75 mmol/L borate buffer (pH 8.5) and 50 microL 20 mmol/L DBD-F in CH3CN were added to the residue and mixed well. The resultant solution was heated for 20 min at 80 degrees C and then cooled in an ice bath. A good separation of DBD-derivatives could be achieved within 45 min using a semi-micro ODS column with an eluent of CH3CN/CH3OH/10 mmol/L imidazole-HNO3 buffer (pH 7.0) (= 45:5:50, v/v/v %). The DBD derivatives were monitored at 565 nm with an excitation at 470 nm. The calibration curves showed good linearity (r = 0.997) with 0.5-15 ng/mL detection limits at a S/N ratio of 3. MDMA and MDA in rat urine could be monitored for 15 h after a single administration of MDMA to rat (2.0 mg/kg, i.p.). The concentrations for MDMA and MDA (n = 3) were 0.13-160.1 and 0.17-10.9 microg/mL, respectively.
authors.affiliation: Department of Clinical Pharmacy, Course of Pharmaceutical Sciences, Graduate School of Biomedical Sciences, Nagasaki University, 1-14 Bunkyo-machi, Nagasaki 852-8521, Japan.
authors.forename: Mitsuhiro
authors.initials: M
authors.lastname: Wada
authors.forename: Shinichi
authors.initials: S
authors.lastname: Nakamura
authors.forename: Mamoru
authors.initials: M
authors.lastname: Tomita
authors.forename: Mihoko N
authors.initials: MN
authors.lastname: Nakashima
authors.forename: Kenichiro
authors.initials: K
authors.lastname: Nakashima
mesh_terms: D015104:3,4-Methylenedioxyamphetamine / Q000652:urine*; D000818:Animals; D001881:Borates; D002138:Calibration; D002851:Chromatography, High Pressure Liquid; D003979:Diazonium Compounds; D016903:Drug Monitoring / Q000379:methods; D005470:Fluorometry; D008297:Male; D018817:N-Methyl-3,4-methylenedioxyamphetamine / Q000008:administration & dosage / Q000652:urine*; D051381:Rats; D017208:Rats, Wistar; D015813:Substance Abuse Detection / Q000379:methods* / Q000592:standards
chemical_list: D001881:Borates; D003979:Diazonium Compounds; C113457:p-(N-N-dimethylamino)-benzendiazonium fluoroborate; D015104:3,4-Methylenedioxyamphetamine; D018817:N-Methyl-3,4-methylenedioxyamphetamine