Development and application of a portable micro liquid chromatograph

Research Project

Project/Area Number	09640723
Research Category	Grant-in-Aid for Scientific Research (C)
Allocation Type	Single-year Grants
Section	一般
Research Field	分離・精製・検出法
Research Institution	KYOTO UNIVERSITY
Principal Investigator	ROKUSHIKA Souji Kyoto University, Science, Lecturer, 大学院・理学研究科, 講師 (80025379)
Co-Investigator(Kenkyū-buntansha)	YAMAMOTO Fumiko Kyoto University, Science, Instructor, 大学院・理学研究科, 助手 (20089139)
Project Period (FY)	1997 – 1999
Project Status	Completed (Fiscal Year 1999)
Budget Amount *help	¥1,200,000 (Direct Cost: ¥1,200,000) Fiscal Year 1999: ¥500,000 (Direct Cost: ¥500,000) Fiscal Year 1998: ¥700,000 (Direct Cost: ¥700,000)
Keywords	HPLC / Portable LC / Micro-column / Nitrous acid / Sugar / UV検出器 / 微量分析 / ポ-タブル液体グロマトグラフ / 紫外検出器
Research Abstract	The purpose of this project is to develop a portable gradient high performance liquid chromatograph and its application and to utilize the micro-column for the analysis of the solute retention mechanisms. The developed instrument is composed of two pumps, a sample injector, a column-heating block and a flow cell in a unit. The newly designed pumps can deliver smooth gradient elution flow without mixing nor dumping devices before the column. The weight of the instrument without recorder was below 11 kg. A zinc discharge lamp was applied to measure the absorbance of the column eluate at 214 nm besides the mercury lamp determined at 254 or 280 nm. At 214 nm, the base line was stable in the 0.05 AUFS range for one year and the detection limit for benzoate esters was as low as 3 ng. Examples of gradient elution chromatography were demonstrated for the chymotripsin digested reduced lysozyme and the PMP (1-phenyl-2-methyl-5-pyrazolone) derivatives of the oligo-sugars yielded by the partial acid digestion of dextrin. The retention behavior of nitrous acid on a C-18 reversed phase column eluting with acidic alcohol-water mixture was studied. The application of the computer simulation for the study of the retention mechanism of the PMP sugar in reversed phase chromatography was reported.