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

诊断用多波长检测高效液相色谱的改进研究

• 批准号: 62490004
• 负责人: DOHI Takeyoshi
• 金额: $ 3.78万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1987
• 资助国家: 日本
• 起止时间: 1987 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-62490004/
• 关键词: high performance liquid chromatography; multiwavelength detection; ultraviolet absorption spectrum; fluorescence emission spectrum; multiwavelength absorption detector; multiwavelength fluorescence detector; biological fluid analysis; 多波長高速液体クロマトグラ

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.

本研究开发了一种新型高效液相色谱多波长检测系统。为了克服二极管阵列探测器在探测波长范围和灵敏度方面的局限性，系统采用光电倍增管作为光学探测器，因为它的灵敏度远大于二极管阵列，其探测波长范围远大于二极管阵列，特别是在短于200 nm的紫外光区域。提出了一种新的机械扫描系统，它是一个带螺旋狭缝的圆盘在光谱仪的像面上旋转。当圆盘旋转时，狭缝的位置在光谱仪衍射的光谱上从左向右移动。通过狭缝的光被收集在光电倍增管上，光电倍增管的输出信号被引入到基于微计算机的信号处理单元。将该系统应用于多波长检测HPLC系统，分离的化合物在紫外区190 nm和250 nm范围内的吸收光谱。通过所开发的系统，氨基酸，糖和羧酸的分析，通过开发的多波长检测HPLC系统。在190 ~ 250 nm波长范围内得到了这些组分的三维色谱图，具有良好的SIN比。所开发的系统也被应用于多波长荧光检测。虽然作为荧光检测器的灵敏度不够，但该检测系统作为HPLC多波长荧光检测系统的可行性得到了证实。