| Project/Area Number |
14340234
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
分離・精製・検出法
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| Research Institution | Kyoto Institute of Technology |
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Principal Investigator |
TANAKA Nobuo Kyoto institute of Technology, Faculty of Textile Science, Professor, 繊維学部, 教授 (60127165)
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| Co-Investigator(Kenkyū-buntansha) |
HOSOYA Ken Kyoto Inst.Tech., Faculty of Textile Sci., Assoc.Professor, 繊維学部, 助教授 (00209248)
MIYABE Kanji Toyama University, Faculty of Engineering, Professor, 工学部, 教授 (10281015)
IKEGAMI Tohru Kyoto Inst.Tech., Faculty of Textile Sci., Instructor, 繊維学部, 助手 (20301252)
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| Project Period (FY) |
2002 – 2004
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| Project Status |
Completed (Fiscal Year 2004)
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| Budget Amount *help |
¥11,800,000 (Direct Cost: ¥11,800,000)
Fiscal Year 2004: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 2003: ¥1,600,000 (Direct Cost: ¥1,600,000)
Fiscal Year 2002: ¥8,500,000 (Direct Cost: ¥8,500,000)
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| Keywords | Liquid chromatography / Peak capacity / Comprehensive 2D-HPLC / HPLC column / High speed separations / Reversed-phase LC / Monolithic silica column / 2D-chromatogram / HPCL充填剤 / 多孔性シリカ連続体 / シリカモノリス / 炭化水素 |
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| Research Abstract |
The objective of this research project was to increase peak capacity of HPLC by developing simple and comprehensive two dimensional (2D) HPLC. By employing a monolithic silica-C18 column at the second dimension (2nd-D), we were able to obtain the following results.
(1) We proved the concept of simple and comprehensive 2D-HPLC in a reversed-phase-reversed-phase mode by using a particle-packed column of fluoroalkyl-bonded silica as a first-dimension (1st-D) column, and C18- or pentabromobenzyloxypropyl-bonded monolithic silica as a 2nd-D column.
(2) By utilizing the difference in selectivity provided by organic solvents, 2D-HPLC was carried out using monolithic silica C18 columns both at 1st-D and at 2nd-D.
(3) 2D-HPLC was carried out by using an ion-exchange column packed with polymer particles at 1st-D and a monolithic silica-C18 column or a monolithic silica capillary column at 2nd-D for peptide separations.
The use of these systems produced (1)peak capacity of up to 1100 in methanol-water mobile phase, (2)peak capacity of 700 or greater for hydrocarbons and benzene derivatives, respectively. The ion-exchange-reversed-phase 2D-HPLC system provided (3)peak capacity of 700 or greater for BSA digest with a short monolithic silica column at 2nd-D, and peak capacity of about 500 for BSA digest by using a 0.2 mmlD, 10 cm long capillary column at 2nd-D, respectively. Optimization of 2D-HPLC conditions were attempted to show the possibility of achieving peak capacity of greater than 2000 in one hour. The flow rate, retention factors of solutes, and column length at 2nd-D were shown to dominate the peak capacity of a 2D-HPLC system. Monolithic silica columns were found to be best suited for 2D-HPLC based on their high permeability and high performance at high linear velocity of mobile phase.
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