| Japanese |
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| Title | ミニカラム法によるビタミンD3分画の分画精製および25(OH)D測定系の検討 |
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| Subtitle | 技術報告 |
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| Authors | 夏目克巳*, 鈴村栄里*, 鈴木智子*, 渡辺幸彦* |
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| Organization | *株式会社エスアールエル |
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| Journal | 核医学 |
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| Volume | 32 |
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| Number | 1 |
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| Page | 99-104 |
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| Year/Month | 1995/1 |
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| Article | 報告 |
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| Publisher | 日本核医学会 |
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| Abstract | 「要旨」C18ミニカラムおよびNH2ミニカラムの2ステップ精製法によるビタミンD代謝体3分画 [25(OH)D, 24,25(OH)2Dおよび1,25(OH)2D] の分画精製について検討した. NH2ミニカラムによる分画精製において, 25(OH)Dはヘキサン:ジクロロメタン (50:50), 24,25(OH)2Dはヘキサン:ジクロロメタン (20:80), 1,25(OH)2Dはヘキサン:イソプロパノール (75:25) で溶出した. この際の該当フラクションへの他代謝体の混入は1.4%以下であった. また血清に添加した3H-25(OH)D, 3H-24,25(OH)2Dおよび3H-1,25(OH)2Dの回収率はそれぞれ73.2±2.45%, 60.0±2.98%, 63.5±3.37% (mean±SD, n=8) であった. さらにミニカラム抽出で得られた25(OH)D画分を用いてCPBAによる測定を検討したところ, 同時再現性CV=4.60〜8.41%, 日差再現性CV=6.62〜16.4%であり, 添加回収試験では81.2〜130%の回収率が得られた. また, 健常人の血清 (5月採取) を測定したところ17.4±6.02ng/ml (mean±SD, n=110) であり, 従来のHPLC精製法の報告と一致した. さらに, 本精製法とHPLC精製法との相関も良好であった (y=-0.38+1.03x, r=0.953, n=36). 25(OH)D測定は, 繁雑なHPLC精製の代わりに簡便なミニカラムを使用した本精製法による測定が可能であると考えられた. |
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| Practice | 臨床医学：一般 |
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| Keywords | Vitamin D metabolites, NH2 cartridge purification |
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| English |
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| Title | Extraction and Purification of the Three Major Vitamin D Metabolites Using C18 and NH2 Cartridges and Measurement of 25-Hydroxyvitamin D |
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| Authors | Katsumi NATSUME, Eri SUZUMURA, Tomoko SUZUKI, Yukihiko WATANABE |
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| Organization | SRL, Inc. |
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| Journal | The Japanese Journal of nuclear medicine |
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| Volume | 32 |
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| Number | 1 |
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| Page | 99-104 |
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| Year/Month | 1995/1 |
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| Article | Report |
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| Publisher | THE JAPANESE SOCIETY OF NUCLEAR MEDICINE |
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| Abstract | [Summary] In this study, the three major vitamin D metabolites : 25-hydroxyvitamin D [25(OH)D], 24,25-dihydroxyvitamin D [24,25(OH)2D], 1,25-dihydroxyvitamin D [1,25(OH)2D] were clearly separated using a NH2 cartridge after acetonitrile and C18 cartridge extraction. In the NH2 cartridge purification procedure, 25(OH)D was eluted with hexane/dichloromethane (50:50), 24,25(OH)2D was eluted with hexane/dichloromethane (20:80) and 1,25(OH)2D was eluted with hexane/isopropanol (75:25). Contamination of each fraction with two other metabolites were less than 1.4%. Recoveries of added 3H-25(OH)D, 3H-24,25-(OH)2D and 3H-1,25(OH)2D were 73.2+-2.45%, 60.0+-2.98% and 63.5+-3.37%, respectively. Using the 25(OH)D fraction after the NH2 cartridge procedure, we measured 25(OH)D using a competitive protein binding assay. The intra- (n=10) and interassay (n=8) coefficients of variation were 4.60-8.41% and 6.62-16.4%, respectively. Analytical recovery of added 25(OH)D was in the range of 81.2-130%. The 25(OH)D values were 17.4+-6.02 ng/ml (mean+-SD) in serum from 110 healthy volunteer collected in May. The correlation of 25(OH)D values was good between cartridge purification and high performance liquid chromatography (HPLC) purification. (y=-0.38+1.03x, r=0.953, n=36) This purification using a simple cartridge procedure was suitable for the measurement of 25(OH)D, and preferable to the time-consuming HPLC purification. |
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| Practice | Clinical medicine |
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| Keywords | Vitamin D metabolites, NH2 cartridge purification |
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