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
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- Satoko Nonaka
- Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
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- Masae Sekine
- Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
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- Makoto Tsunoda
- Graduate School of Pharmaceutical Sciences; University of Tokyo; Tokyo Japan
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- Yuji Ozeki
- Department of Psychiatry; Dokkyo Medical University School of Medicine; Tochigi Japan
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- Kumiko Fujii
- Department of Psychiatry; Dokkyo Medical University School of Medicine; Tochigi Japan
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- Kazufumi Akiyama
- Department of Biological Psychiatry and Neuroscience; Dokkyo Medical University School of Medicine; Tochigi Japan
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- Kazutaka Shimoda
- Department of Psychiatry; Dokkyo Medical University School of Medicine; Tochigi Japan
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- Takemitsu Furuchi
- Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
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- Masumi Katane
- Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
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- Yasuaki Saitoh
- Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
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- Hiroshi Homma
- Faculty of Pharmaceutical Life Sciences; Kitasato University; Tokyo Japan
書誌事項
- タイトル別名
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- 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.
収録刊行物
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- Journal of Separation Science
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Journal of Separation Science 37 (16), 2087-2094, 2014-06-30
Wiley