Studies on improvement of multiwavelength detection high performance liquid chromatography for diagnostic

1989 Fiscal Year Final Research Report Summary

• Project/Area Number: 62490004
• Research Category: Grant-in-Aid for General Scientific Research (B)
• Allocation Type: Single-year Grants
• Research Field: 広領域
• Research Institution: The University of Tokyo
• Principal Investigator: DOHI Takeyoshi The Univ. of Tokyo, Fac. of Eng., Professor, 工学部, 教授 (40130299)
• Co-Investigator(Kenkyū-buntansha): SAKUMA Ichiro Tokyo DENKI University, Fac. of Sci. and Eng. Research Associate, 理工学部, 助手 (50178597) ; TAKAI Nobuharu The Univ. of Tokyo, Inst. of Industrial Sci. Associate Professor, 生産技術研究所, 助教授 (20013188) ; OHKUBO Akiyuki The Univ. of Tokyo, Fac. of Medicine, Professor, 医学部, 教授 (20010423) ; SENO Manabu The Univ. of Tokyo, Inst. of Industrial Sci. Professor, 生産技術研究所, 教授 (40013099)
• Project Period (FY): 1987 – 1989
• Keywords: high performance liquid chromatography / multiwavelength detection / ultraviolet absorption spectrum / fluorescence emission spectrum / multiwavelength absorption detector / multiwavelength fluorescence detector / biological fluid analysis

Research Abstract

In this study, a new type of a multiwavelength detection system for high performance liquid chromatography was developed. To overcome the limitations of diode array detectors about their detectable wavelength range and sensitivity, the system adopted a photomultipliers as an optical detector since its sensitivity is much greater than that of a diode array and its detectable wavelength range is much wider than that of a diode array, especially in the region of ultra-violet light shorter than 200nm. A new mechanical scanning system was developed where a disk with a spiral shaped slit is rotating at the image plane of the spectrometer. The position of the slit moves from left to right on the spectrum diffracted by the spectrometer as the disk rotates. Light passing through the slit is collected on the PMT and PMT output signal is introduced to a microcomputer based signal processing unit. The developed system was applied to the multiwavelength detection HPLC system where the absorption spectra of separated compounds in the region from 190nm and 250nm in ultraviolet region. By the developed system, amino acids, sugars and carboxylic acids were analyzed by the developed multiwavelength detection HPLC system. Three dimensional chromatograpms of these components in the region from 190nm to 250 nm were obtained with good SIN ratio. The developed system was also applied to multiwavelength fluorescence detection. Although the sensitivity was not enough as a fluorescence detector, the feasibility of the detection system as an multiwavelength fluorescenece detection system for HPLC was confirmed.