Abstracts of Oral and Poster Presentations

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  • I. TOXIC0L.-TOXIN REVIEWS, 17(1), 85-98 (1998)

    ABSTRACTS OF ORAL AND POSTER PRESENTATIONS

    AUTOMATED PREPARATION AND ANALYSIS OF MORPHINE IN BIOLOGICAL E U I D S

    A l a n Namera*, Miluo Yashiki*, Kanako Okada*, Yasumasa Iwasakr*, Tohru Kojima* and HaJirne Kawakami**, *Department of Legal Medicine, Hiroshima University School of Medicine, 1-2-3, Kasumi, Minamr ku, Hiroshima 734, Japan and **Yokogawa Analqtical Systems Inc , 1-15-5, Naka-cho, Musashini 180, Japan

    A method for the automatic analqsis of morptune i n biological fluids u a s debeloped using a PrepStation (Hewlett Pachard, USA) in combination with a gas chromatographimass spectrometer Human interaction is only required to place samples on the auto-sampler tray Recoveq of morphine was more than 90% The calibration curve, using the internal standard method, demonstrated good lineanty throughout the concentration range from 0 01 to 1 0 [ L gjml

    for unne and from 0 05 to 1 0 u g/ml for serum The proposed method was applied to some clinical cases

    MICROANALYSIS OF C A N N A B I S COMPONENTS A N D THEIR METABOLITES BY ELISA. K. Watanabe, Y. Tateoka, T. Matsunaga, I. Yamamoto), Y. Shoyama & H. Yoshimura), ) Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa 920- 11, Japan, *) Department of Pharmacognosy, Faculty of Pharmaceutical Sciences, Kyushu Uiversity, Fukuoka 812, Japan & 3, Department of Food and Nutrition, Nakamura Gakuen University, Fukuoka 814-01, Japan

    The monoclonal antibody (MAb-4A4) against A9-tetrahydrocannabinolic acid (A9-THCA) was produced in mice immunized with A9-THCA-BSA conjugate. Competitive ELISA was established using MAb-4A4, A9-THCA-@alanineBSA conjugate, peroxidase- or alkaline phosphatase-conjugated anti-mouse IgG. The antibody cross-reacted with THC, cannabidiol, cannabinol and 15 kinds of THC metabolites, but not with other biological

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  • 86 ABSTRACTS

    compounds such as cholesterol, testosterone and anandamide. Thus, the ELISA was not specific for A9-THCA but detected various cannabinoids indicating the advantage of the antibody for screening of cannabinoid related compounds. This system is applicable to detection of cannabinoids and their metabolites in biological samples.

    A NEW SCREENING METHOD FOR AMPHETAMINE AND METHAM-

    PHETAMINE USING DANSYL CHLORIDE DERIVATIZATION. H. Y A M A D A I . H. IWASAKI ' , K. OGURI' & S. WADA*. I Faculty of Pharmaceutical Sciences, Kyushu University 62, Fukuoka 8 12-81, Japan; Kyushu District Narcotic Control Office, Fukuoka 8 12, Japan.

    A new screening method for amphetamines was developed. It consists of derivati- zation with dansyl chloride, extraction of the derivative using a Sep-Pak cartridge, a reversed phase column, and visualization of the fluorescence of the cartridge. Of fifty five compounds examined, amphetamine, methamphetamine and the methylene- dioxy derivatives exhibited strong fluorescence, while related compounds, such as N- ethylamphetamine and fenetylline, were negative or weak positive. Amino acids were negative. In consistent with this, drug-free control urine exhibited no fluores- cence in cartridge. The disadvantage of the present method is that i t is a multi- step procedure and needs 20 - 30 min for screening. However. since the method has a different specificity from the widely used immunochemical method, i t is suggested as a specific screen for amphetamines.

    SIMULTANEOUS DETERMINATION OF METHAMPHETAMINE AND p-HYDROXY DESMETHYL BENZPHETAMINE BY HPLC USING POST- COLUMN SIMON'S REACTION Laboratory, Ishikawa Pref Police H Q . 2-1-1 Hirosaka, Kanazawa 920 Japan

    S Chinaha & N Takayama, Forensic Science

    A high-performance liquid chromatographic method for the sitnultaneous determination of methamphetamine (MA) and p-hydrox) desrnethyl benzphetamine (OHnorBZP) using post-column Simon's reaction (Simon's-HPLC) has been developed MA is used by most stimulant addicts Benzphetamine (BZP), from which OHnorBZP or MA etc are biotransformed, is also being irnponed or used as street drug The detection limit of MA was Ing This data indicated that the Simon's-HPLC has about 100 times sensitivity in comparison with the thin-

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  • ABSTRACTS 87

    l q e r chromatography using Simons reaction (Simons-TLC) The detection liniit of OHnorBZP ivas ca.20ng. The calibration curve of MA \$as linear at the range 0 1 - 1 000pgiiiil. and the intermediate precisions of within-run and between-run assals Mere I 1% and 2 12%. respectively. MA i n 13 stimulant addicts urine sainplcc was determined by the Simons-HPLC and gas chromatography with flame ionization dctection. and these quantiiative values Mere good correlative M A and OIHiiorBZP i n the MA users urine spiked OHnorBZP could be analyzed simultaneously M ithout interference peaks. Our method was useful as a substitute of the Slmoils- ILC.

    SENSITIVE DETERMINATION OF THINNER COMPONENTS IN HUMAN BODY FLUIDS BY CAPILLARY GAS CHROMATOGRAPHY WITH LOW OVEN

    TEMPERATURE. X.-P. Lee, T. Kumazawa, K. Satoa), K. Watanabeb, H. Senob and 0.

    Suzuki. Department of Legal Medicinc, Showa University School of Medicine, 1-58

    Hatanodai, Shinagawa-ku, Tokyo 142, Japan ; bDepartment of Legal Medicine, Hamamatsu

    University School of Mcdicine, 3600 Handa-cho, Hamamatsu 431 -31, Japan.

    A sensitive method is presented for determination of thinner components in human body

    fluids by capillary gas chromatography (GC) with low oven temperature for trapping headspace

    vapor insidc the capillary column. After heating a whole blood or urinc sample containing

    ethyl acetatc, bcnzene, ri-butanol, toluenc, rr-butyl acetate, n-isoamyl acetate and ethyl benzene

    as internal standard in a 7.5 ml-vial at YO C for 30 min, 5 ml of headspace vapor was drawn

    into a glass syringe. All vapor was introduced through an injection port in the splitless mode

    into a D E h 2 4 middle-bore capillary column with the oven temperature at 5 C for trapping

    all volatilc compounds, and the oven temperature was programmed up t o 110 C for detection

    of the compounds by GC with a flame ionization detector. The prcsent conditions gave sharp

    peaks and good separation of each compound with low background noises. Recoveries of thc

    6 compounds were 4.7&75.7 o/o and 3.SCA1.7 % for whole blood and urine samples,

    respcctively. The CV values of all compounds in whole blood and urine samples were

    3.5-9.5 %. The calibration curves showed linearity in the range 0.78400 ngi0.5 ml wholc

    blood or urine; the detection limits were 0.5-5 ngj0.5 ml. The data on actual analysis of

    toluene and n-butyl acetate in postmortem blood were also reported.

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  • 88 ABSTRACTS

    SENSITIVE DETERMINATION OF CHLOROFORM AND METHYLENE CHLORIDE IN WHOLE BLOOD BY CAPILLARY GAS CHROMATOGRAPHY WITH CRYOGENIC OVEN TEMPERATURE Kanako Watanabe, Akira Ishii, Hiroshi Sen0 & Osamu Suzuki, Department of Legal Medicine, Hamamatsu University School of Medicine, 3600 Handa- cho, Hamamatsu 43 1-3 1, Japan & Hideki Hattori, Department of Legal Medicine, k c h i Medical University, Nagakute-cho, Aichi, 480- 1 1, Japan

    A new and sensitive method of gas chromatography (GC), for measurements of chloroform or methylene chloride in whole blood with use of cryogenic oven temperature, is presented. After heating a blood sample containing chloroform and methylene chloride (internal standard, vice versa) in a 7.0-ml vial at 55C for 20 min, 5 ml of the headspace vapor was drawn into a glass

    syringe. All vapor was introduced into an Rtx-Volatiles middle-bore capillary column in the splittless mode at -30C of oven temperature for trapping entire amounts of compounds to be analyzed, and the oven temperature was programmed up to 280% for detection of the compounds and for cleaning of the column. The present conditions gave sharp peaks for hoth chloroform and methylene chloride, and very low background noises for whole blood samples. As much as 1 1 . 5 and 20 0 YO of chloroform and methylene chloride, respectively, which had been added to whole blood in a vial, could be introduced into the GC column. The calibration curves showed linearity in the range of 0.05-5.0 mg/0.5 ml whole blood. The detection limit was about 2 ng0 .5 ml. The coefficients of intra-day and inter-day variations of area ratios of chloroform / methylene chloride, which had been spiked to human blood, were not greater than 1.74 and 8 85 %, respectively. The data on chloroform or methylene chloride in rat blood after inhalation of each compound were also presented

    ANALYSIS FOR PESTICIDE COMPONENTS BY PULSE HEATlNG - GAS CHROMATOGRAPHY MASS SPECTROMET'RY. T. Takayasu, T. Ohshima, T. Kondo, M. Ohtsuji & Y. Sato, Department of Legal Medicine, Kanazawa University Faculty of Medicine, School of Medicine. 13-1 Takara-machi, Kanazawa 920-8640. Japan

    Analysis fo

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