| Project/Area Number |
05454580
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| Research Category |
Grant-in-Aid for General Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Research Field |
応用薬理学・医療系薬学
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| Research Institution | TOHOKU UNIVERSITY |
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Principal Investigator |
HISHINUMA Takanori Tohoku University Hospital, AssociateProfessor, 医学部・附属病院, 助教授 (20199003)
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| Co-Investigator(Kenkyū-buntansha) |
MIZUGAKI Michinao Tohoku University, Hospital, Professor, 医学部・附属病院, 教授 (60004595)
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| Project Period (FY) |
1993 – 1994
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| Project Status |
Completed (Fiscal Year 1994)
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| Budget Amount *help |
¥6,100,000 (Direct Cost: ¥6,100,000)
Fiscal Year 1994: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 1993: ¥5,100,000 (Direct Cost: ¥5,100,000)
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| Keywords | prostaglandin / prostacycline / thromboxane / GC-MS / 11-dehydro-TXB2 / 11-dehydro-TXB3 / biodistribution / 11-dehydro-TXB_3 / 6,15-diketo-13,14-dihydro-PGF_<1α> / GC / MS / SIM / モノクローナル抗体 / 阻害ELISA / 3型PG |
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| Research Abstract |
1.The microdetermination of 11-dehydro-thromboxane B3 (11-dehydro-TXB3) in human urine is described. We prepared [^<18>O] 11-dehydro-thromboxane B3 for use as an internal standard (IS). Samples to which was added [^<18>O] analogue were extracted by chromatographic sample preparation using Sep Pak tC18 and silica gel column. Conversion of the extracted11-dehydro-thromboxane B3 to 1-methyl ester (ME) -11-n-propylamide (PA) -methoxime-9,11,15-tris-dimethylisopropylsilyl (DMIPS) ether derivative was followed by gas chromatography/high resolution-selected ion monitoring (GC/HR-SIM). Interfering substances from the urine matrix were eliminated during GC/HR-SIM analysis using a MP-65HT column. Good linear response over the range of 10 pg-100 ng/tube was demonstrated. We were able to detect 11-dehydro-TXB3 in the range from 26 to 375 pg/ml of the human urine. The present method can be applied to the determination of 11-dehydro-TXB3 in the human urine
2.The microanalysis of 2,3-dinor-6-keto-pros
… More
taglandin F1alpha (I) in human urine was developped. Samples to which was added [^2H4] -analogue as an internal standard were extracted by chromatographic sample preparation using Bond Elut C18, and silica gel. Conversion of the extracted I to 1-methyl ester-6-methoxime-9,11,15-tris-dimethy-lisopropylsilyl ether derivative was followed by gas chromatography/high resolution-selected ion monitoring (GC/HR-SIM). Inrerfering substances from the urine matrix were eliminated during GC/HR-SIM analysis.
A method of the microdetermination of 2,3-dinor-DELTA^<17>-6-keto-PGF1alpha, a urinary metabolite of PGI3, is developped. An authentic 2,3-dinor-DELTA^<17>-6-keto-PGF1alpha was preparedfrom DELTA^<17>-6-keto-PGF1alpha incubated with homogenate of rat liver. [^<18>O] DELTA^<17>-6-keto-PGF1alpha was synthesized by repeating base-catalyzed hydrolysis of methyl ester derivatives in [^<18>O] water, to obtain an internal standard in gas chromatography/selected ion monitoring (GC/SIM) of DELTA^<17>-6-keto-PGF1alpha. Good linear response over the range of 10 pg-10ng was demonstrated. Chromatographic conditions using a MP-65HT column presented nearly baseline separation of DELTA^<17>-6-keto-PGF1alpha and 6-keto-PGF1alpha. We were able to detect DELTA^<17>-6-keto-PGF1alpha in the range from 6 to 26 pg/ml of the human plasma. The present method can be applied to the determination of DELTA^<17>-6-keto-PGF1alpha in the human urine and plasma. Less
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