Simultaneous determination of<i>N</i><sup>G</sup>-monomethyl-<scp>l</scp>-arginine,<i>N</i><sup>G</sup>,<i>N</i><sup>G</sup>-dimethyl-<scp>l</scp>-arginine,<i>N</i><sup>G</sup>,<i>N</i><sup>G′</sup>-dimethyl-<scp>l</scp>-arginine, and<scp>l</scp>-arginine using monolithic silica disk-packed spin columns and a monolithic silica column

  • Satoko Nonaka
    Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
  • Masae Sekine
    Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
  • Makoto Tsunoda
    Graduate School of Pharmaceutical Sciences; University of Tokyo; Tokyo Japan
  • Yuji Ozeki
    Department of Psychiatry; Dokkyo Medical University School of Medicine; Tochigi Japan
  • Kumiko Fujii
    Department of Psychiatry; Dokkyo Medical University School of Medicine; Tochigi Japan
  • Kazufumi Akiyama
    Department of Biological Psychiatry and Neuroscience; Dokkyo Medical University School of Medicine; Tochigi Japan
  • Kazutaka Shimoda
    Department of Psychiatry; Dokkyo Medical University School of Medicine; Tochigi Japan
  • Takemitsu Furuchi
    Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
  • Masumi Katane
    Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
  • Yasuaki Saitoh
    Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
  • Hiroshi Homma
    Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan

書誌事項

タイトル別名
  • Liquid Chromatography

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説明

We have developed and validated a high-performance liquid chromatography method that uses monolithic silica disk-packed spin columns and a monolithic silica column for the simultaneous determination of N(G)-monomethyl-L-arginine, N(G),N(G)-dimethyl-L-arginine, and N(G),N(G')-dimethyl-L-arginine in human plasma. For solid-phase extraction, our method employs a centrifugal spin column packed with monolithic silica bonded to propyl benzenesulfonic acid as a cation exchanger. After pretreatment, the methylated arginines are converted to fluorescent derivatives with 4-fluoro-7-nitro-2,1,3-benzoxadiazole, and then the derivatives are separated on a monolithic silica column. L-arginine concentration was also determined in diluted samples. Standard calibration curves revealed that the assay was linear in the concentration range 0.2-1.0 μM for methylated arginines and 40-200 μM for L-arginine. Linear regression of the calibration curve yielded equations with correlation coefficients of 0.999 (r(2)). The sensitivity was satisfactory, with a limit of detection ranging from 3.75 to 9.0 fmol for all four compounds. The RSDs were 4.3-4.8% (intraday) and 3.0-6.8% (interday). When this method was applied to samples from six healthy donors, the detected concentrations of N(G)-monomethyl-L-arginine, N(G),N(G)-dimethyl-L-arginine, N(G),N(G')-dimethyl-L-arginine and L-arginine were 0.05 ± 0.01, 0.41 ± 0.07, 0.59 ± 0.11, and 83.8 ± 30.43 μM (n = 6), respectively.

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