| Project/Area Number |
13672250
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Physical pharmacy
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| Research Institution | The University of Tokyo |
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Principal Investigator |
SANTA Tomofumi The University of Tokyo, Graduate School of Pharmaceutical Sciences, Associate Professor, 大学院・薬学系研究科, 助教授 (30187306)
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| Co-Investigator(Kenkyū-buntansha) |
IMAI Kazuhiro The University of Tokyo, Graduate School of Pharmaceutical Sciences, Professor, 大学院・薬学系研究科, 教授 (50012620)
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| Project Period (FY) |
2001 – 2002
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| Project Status |
Completed (Fiscal Year 2002)
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| Budget Amount *help |
¥3,700,000 (Direct Cost: ¥3,700,000)
Fiscal Year 2002: ¥1,800,000 (Direct Cost: ¥1,800,000)
Fiscal Year 2001: ¥1,900,000 (Direct Cost: ¥1,900,000)
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| Keywords | nitrogen oxide / HPLC / fluorescence / dimethylarginine / OPA / derivatization |
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| Research Abstract |
A fully automated analyzer for methylated L-arginine metabolites (N,N-dimethyl-L-arginine (ADMA), N-methylarginine (NMMA) and N, N'-dimethyl-L-arginine (SDMA)) by high-performance liquid chromatography with post-column fluorescence derivatization was developed. This system consists of an in-line extraction, a separation on a reversed phase ion-pair chromatography, a post-column derivatization by o-phthaladehyde (OPA) and thiol reaction, and fluorescence detection. NMMA, ADMA and SDMA were separated in 40 min with isocratic elution by a combination of octanoate and cyclohexanecarboxylate as ion-pair reagents. The eluate was monitored at 450 nm with the excitation at 337 nm. The calibration curves for NMMA, ADMA and SDMA showed linearity over the range from 0.05 μmol 1^<-1> (0.5 pmol on column) to 5.0 μmol 1^<-1> (50 pmol on column). This method does not require any time-consuming pre-treatment and requires only 10-μl of plasma sample for assay.
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