線上微透析取樣配合原子吸收光譜連線分析系統之研究及其生物醫學應用

Study on the Hyphenated System of On-Line Microdialysis Sampling Coupled with Atomic Absorption Spectrometry and Its Biomedical Applications

曾維昌(Wei-Chang Tseng)

指導教授 ： 黃友利 ； 陳田柏

高雄醫學大學/醫學院/醫學研究所/博士(2003年)

摘要

本研究目的在於發展線上微透析取樣技術(MDS)、流動注入系統(FI)與各式原子吸收光譜(AAS)之連線分析方法，經嚴謹的驗證程序，實際分析活體動物內或離體生物樣品中的微量元素及其物種，以確定連線方法之可行性。本研究的內容主要有三部分：第一部分，建立完全自動化之線上微透析取樣系統、高效液相層析、流動注入系統與氫化物產生原子吸收光譜(MDS-HPLC-FI-HGAAS)之連線分析方法，直接測定尿中四個砷物種(三價砷、五價砷、單甲基砷和雙甲基砷)，並利用此自動化之連線分析系統，將微透析探針植入活體大白鼠膀胱內，進行尿中砷物種分析。第二部分，發展線上微透析取樣系統、流動注入系統與電熱式原子吸收光譜(MDS-FI-ETAAS)之連線分析方法，本研究於微透析取樣系統和電熱式原子吸收光譜之間，設計一套可導入適量樣品的流動注入介面，使透析液與修飾劑在線上混合並導入石墨管，此連線分析系統經分析效能評估後，可連續監測活體大白鼠腦中錳的分佈。第三部分，發展線上微透析取樣技術、流動注入系統和火焰式原子吸收光譜(MDS-FI-FAAS)之連線分析方法，此方法利用簡單設計的流動注入介面，使收集的透析液被攜帶溶液導入火焰原子化器，毋需任何樣品前處理步驟，即可直接和快速地測定人類全血樣品中可被透析的鈣。本研究研發的三種線上微透析與原子吸收光譜連線分析方法，將有助於進一步研究微量元素之生物醫學機轉。

關鍵字

線上微透析取樣 ； 流動注入 ； 原子吸收光譜 ； 連線分析方法 ； 生物醫學應用

並列摘要

The hyphenated techniques of microdialysis sampling (MDS) coupled on-line with flow injection (FI) atomic absorption spectrometry (AAS) have been developed in the present study. The performance characteristics (linearity, detection limit, spiked recovery, precision and accuracy) of the on-line MDS coupled with AAS system were carefully validated. The determination and speciation of trace elements in in vivo living animals or in vitro biological fluids were performed by the proposed on-line MDS-FI-AAS system. Three major works were carried out in this study. Firstly, an on-line and fully automated MDS-HPLC-FI-HGAAS system has been developed to determine four urinary arsenic species [arsenite (AsIII), arsenate (AsV), monomethylarsonic acid (MMA) and dimethylarsinic acid (DMA)]. The continuous and dynamic in vivo monitoring of four urinary arsenic species in bladders of anesthetized living rats were investigated using the proposed on-line method. In the subsequent part, an on-line MDS coupled with electrothermal atomic absorption spectrometry (ETAAS) has been developed by a home-made FI interface. The use of an on-line MDS-FI-ETAAS system permitted the in situ, dynamic and continuous in vivo monitoring of extracellular manganese in brains of anaesthetized living rats. Finally, a direct and rapid determination of diffusible calcium in human whole blood using on-line MDS coupled with FI flame atomic absorption spectrometry (FAAS) has been developed. The use of an on-line MDS-FI-FAAS system makes it possible to rapidly determine diffusible calcium in complex matrix of human whole blood without sample pretreatment. These on-line MDS-FI-AAS methods proposed in the present study may benefit the further researches into the biomedical mechanisms of trace elements.

並列關鍵字

on-line microdialysis sampling ； flow injection ； atomic absorption spectrometry ； hyphenated analytical method ； biomedical application

參考文獻

第1章

[1]K. J. McLaughlin, A. A. Faibushevich and C. E. Lunte, Analyst, 2000, 125, 105.

[2]J. E. Thompson, T. W. Vickroy and R. T. Kennedy, Anal. Chem., 1999, 71, 2379.

[3]F. Xiang, Y. Lin, J. Wen, D. W. Matson and R. D. Smith, Anal. Chem., 1999, 71, 1485.

[4]M. Pravda, L. Bogaert, S. Sarre, G. Ebinger, J. M. Kauffmann and Y. Michotte, Anal. Chem., 1997, 69, 2354.