| Project/Area Number |
16350045
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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 |
Analytical chemistry
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
HIROKAWA Takeshi Hiroshima University, Graduate School of Engineering, Professor, 大学院・工学研究科, 教授 (30116652)
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| Co-Investigator(Kenkyū-buntansha) |
IKUTA Natsuki Hiroshima University, Graduate School of Engineering, Research associate, 大学院・工学研究科, 助手 (00274118)
ITO Kazuaki Kinki Univ., Faculty of Engineering, Associate Professor, 工学部, 助教授 (80151497)
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| Project Period (FY) |
2004 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥9,700,000 (Direct Cost: ¥9,700,000)
Fiscal Year 2005: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2004: ¥7,100,000 (Direct Cost: ¥7,100,000)
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| Keywords | Microchip electrophoresis / On-line preconcentration / Transient isotachophoresis / Highly sensitive analysis |
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| Research Abstract |
Microchip electrophoresis (MCE) is attracting a great deal of attention in the field of bio-analysis for DNA fragments and proteins because of downsizing of the electrophoretic apparatus, rapid automated analysis and parallel processing. However the separability of conventional MCE procedure is not high because of its geometric restriction (for example, the separation channel of a commercially available microchip is less than 30mm) and its analytical sensitivity is also restricted due to the small volume of the sample reservoir (ca.3μl). The reason of the latter issue is due to the character of pinched injection procedure, which introduces only a part of analytes from the reservoir. For example, when DNA fragments are analyzed by the conventional method using UV detection, the LOD is a few hundreds of μg/mL. We developed a new injection method named "electrokinetic supercharging" (a combined technique of electrokinetic injection and transient Isotachophoresis, abbreviated as EKS), which enables preconcentrating injection to achieve high-sensitive analysis. We demonstrated 20-30 times enhancement of the LOD in the analysis of DNA fragments and proteins, even when commercially available MCE apparatus and microchip are used. By applying computer simulation (CFD-ACE+, USA), a new microchip was designed with a preconcentration channel suitable for EKS procedure. The simulation was very effective in designing the geometry of the bent channel and evaluation of the analyte dispersion at the cross part of the channels. The developed microchip and analytical method does not conflict with the Ramsey patent, which is the basic patent of the conventional pinched injection. We have applied a new patent (Microchip Electrophoresis Method and Apparatus) to the government of Japan (2005) and USA (in preparation, 2006) as a collaborative work of Hiroshima University and Shimadzu Corporation.
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